Although the literature reports many deleterious effects associated with maternal cigarette smoking on the fetus, the molecular alterations and mechanisms of action are not yet clear. Smoking may act directly on nuclear DNA by inducing mutations or epigenetic modifications. Recent studies also indicate that smoking may act on mitochondrial DNA by inducing a change in the number of copies to make up for the damage caused by smoking on the respiratory chain and lack of energy. In addition, individual genetic susceptibility plays a significant role in determining the effects of smoking during development. Furthermore, prior exposure of paternal and maternal gametes to cigarette smoke may affect the health of the developing individual, not only the in utero exposure. This review examines the genetic and epigenetic alterations in nuclear and mitochondrial DNA associated with smoke exposure during the most sensitive periods of development  and assesses how such changes may have consequences for both fetal growth and development.Newborns exposed to maternal cigarette smoke (CS)  The disease risks from cigarette smoking increased over most of the 20th century . As a main utero and perinatal. Even if the two latter periods have been most extensively studied, it is well known that maternal lifestyle and environmental exposures during these three periods can lead to adverse health effects throughout the life course. For instance, in utero exposure to cigarette smoke is associated with reduced fecundity in females and with altered reproductive development in males [Cigarette smoke, with its mix of hazardous chemicals, carcinogens, pulmonary irritants, cilia toxicants, cardiotoxin and teratogens, has been associated with childhood diseases and lifelong risks of chronic disease ,5 pre-tein males  and cigain males . Regardiin males ,13.The critical role of early life events that increase susceptibility to certain chronic diseases resides in the developmental plasticity feature of mammalian organisms. This plasticity consists in the modulation of gene expression during early development by environmental and intrinsic characteristics. Developmental plasticity is mediated, at least in part, by epigenetic processes such as DNA methylation and histone modifications that lead to changes in gene expression . TherefoIn this review, we analyze the genetic and epigenetic alterations in nuclear and mitochondrial DNA associated with smoke exposure during the prenatal and early postnatal periods of development, covering the most crucial findings and revising the literature related to these critical epigenetic modifications.Exposures to environmental contaminants and lifestyle choices during gestation and/or shortly after birth can significantly influence individual phenotype and susceptibility to disease during childhood and throughout the life course. Moreover, environmental exposures also represent the major source of mutation and DNA damage that threaten the integrity of the genome, at gene and chromosome levels.in utero exposure [in vitro study on fetal cells (amniocytes) treated with nicotine, Demirhan and colleagues found an increase in chromosomal aberrations (CAs) in cells exposed to nicotine [in utero exposure to cigarette smoke or environmental tobacco smoke (second-hand smoke) led to a significant increase in DNA damage. They also reported a direct association between subject with serious DNA damage and low birth weight of neonates, an indicator of health problems later in adult life [The mutagenic impact of maternal cigarette smoking on fetal DNA has been described in several studies . In adulexposure . In an inicotine . In vivonicotine ,24,25. Ault life .Therefore, there is a large body of evidence that demonstrates the direct genotoxic effects of smoke components, namely nicotine and its metabolized derivatives, all along the life course, as well as during embryonic development.CYP1A1), glutathione S-transferase (GST), and N-acetyltransferases (NAT) [CYP1A1), while phase II enzymes, including NAT2, GSTM1 and GSTT1, catalyze the activation or deactivation of aromatic and heterocyclic amine carcinogens and intermediate products generated by the CYP1A1 enzyme. Each of these enzymes exhibits SNPs that have been shown to be related to adverse pregnancy outcomes and specific cancers [CYP1A1 and GSTT1 genotypes modified the association between maternal cigarette smoking and infant birth weight. Wu and colleagues [CYP1A1 and GSTT1, and DNA damage. Other studies have confirmed variable DNA damage caused by smoking in genes that regulate genotoxic metabolism [Not all mothers who smoke have children with consequences such as low birth weight or intrauterine growth restriction. The reason for this variability is largely unknown, but it may be related to individual genetic susceptibility. Studies that examine genetic polymorphisms associated to tobacco smoke exposure reveal variability in the frequency of single nucleotide polymorphysms (SNPs) in genes associated with the metabolism of polycyclic aromatic hydrocarbons (PAHs) and aromatic amines. The genes with the highest frequency of SNPs associated to cigarette smoke exposure are isoenzyme of cytochrome P450 (es (NAT) ,29,30,31 cancers ,27,28. Wlleagues  also foutabolism ,33, althTherefore, there are indications that cigarette smoke toxicity depends not only on the intrinsic characteristics of the toxic components in the environmental stress, but also on the individual susceptibility of the subjects exposed to cigarette smoke.in utero and early childhood may influence life-long susceptibility to disease via epigenetic alterations.Epigenetics provide a window through which we can understand the impact of the environment, nutrition and lifestyle choices on chronic disease susceptibility and risk, since the very early life . TransieEpigenetic regulation is achieved by a combination of several molecular machineries, among which there are DNA methylation, histone modification and the noncoding RNA-mediated gene regulation, especially by microRNA (miRNA) .DNA methylation is one of the most widely studied and well-characterized marks of epigenetic regulation and consists of both specific DNA promoter methylation and global DNA methylation in non-coding areas of the genome. Interestingly, both of them are commonly found altered in most types of cancer  and emerGlobal DNA methylation has been proposed as an early epigenomic mark in the initial transition from a normal to a diseased cell . TypicalGlobal loss of DNA methylation is a complex phenomenon that is not clearly understood. What is known is that huge decrease in methylation of large regions enriched with cytosines  cannot be due only to changes in the methylation of gene-coding regions, because these account for less than 5% of cytosines found in human DNA. Rather, global DNA hypomethylation is due to changes in the methylation of CpG rich non-coding areas of the genome, such as satellites SAT2 and SAT3, and interspersed repeat sequences such as long interspersed elements (LINEs), short interspersed elements (SINEs) and long terminal containing repeats (LTRs). These changes may well be adaptive responses, evolution-conserved, that maintain homeostasis and assure cell survival in the face of threatening and noxious environmental stimuli.Because of the increasing evidence that differential DNA epigenome is linked to human disease, several investigators have studied the association between environmental exposure and epigenetic mechanisms such as tobacco smoke exposure and DNA methylation, especially in the early life period.Terry and colleagues  studied in utero smoke exposure is associated with lower levels of global DNA methylation and higher level of methylation of single CpG loci in eight genes analyzed. Interestingly, another independent study confirmed that global DNA methylation in newborns is associated with exposure to prenatal maternal smoking by measuring global DNA methylation through examination of the cord blood serum of full term babies [In two different studies ,43, Bretm babies . The stuin utero exposure to tobacco smoke with gene-specific DNA methylation alterations in the placenta. Suter and colleagues observed that maternal tobacco use is associated with alterations in promoter methylation of placental CYP1A1 gene and that these changes are correlated with CYP1A1 expression and fetal growth restriction [Interestingly, several studies correlate triction . Using Itriction . The strtriction .CYP1A1 gene and smoke exposure has been confirmed. Indeed, Joubert and colleagues observed a strong dose dependent association between maternal smoking during pregnancy and the methylation of genes involved in the pathway for the detoxification of xenobiotics in tobacco smoke such as CYP1A1 and aryl-hydrocarbon receptor repressor (AHRR) together with a trascriptional repressor gene growth factor independent 1 transcription repressor (GFI1) that is involved in diverse developmental process [CYP1A1 AHRR and GFI1 persist also during adolescence [Interestingly, in a recent genome-wide methylation study conducted on umbilical cord blood samples from a Norwegian Mother and Child Cohort study (MoBa), the association between methylation of  process . Moreovelescence .in utero tobacco exposure and methylation levels of a parentally imprinted gene has been characterized. Indeed, Murphy and colleagues have shown that prenatal exposure to cigarette smoke can cause increased methylation of differentially methylated regions (DMR) in the paternally expressed Insulin-Like Growth Factor II (IGF2) gene, from the analysis of umbilical cord blood of 418 pregnant women. They showed that higher methylation of the IGF2 DMR was associated with low birth weight, predominantly in male infants [IGF2 plays an important role in regulating growth [Additionally, a significant association between  infants . IGF2 plg growth  and is ain utero is linked to behavioral and cognitive problems [BDNF) in blood samples from adolescents whose mothers smoked during pregnancy. This gene is important for long-term memory, it helps to support survival of existing neurons and encourages the growth and differentiation of new neurons and synapses [BDNF appears to be very susceptible to methylation by environmental factors, especially in exon 6, but other preliminary studies identified methylation changes upstream of exon 1 as potential diagnostic blood biomarkers for major depression [Exposure to tobacco smoke problems . Some stsynapses . BDNF appression . and placental and embryonic development . Moreover, the most remarkable observation in their data is the identification of unique smoking-related methylation changes in adults versus newborns, suggesting potential age-related differences in the response to exposure. These differences may be related to age-related metabolic and detoxification rates, mediated by epigenetic susceptibility to environmental stress.Finally, a very interesting study has been recently published by the group of Wilcox  that perin utero smoking exposure by self-report, others use air monitoring of nicotine concentration, or measure the levels of cotinine, a nicotine metabolite, in blood or urine. Finally, other studies are yet limited by not taking into account the impact of second-hand smoke exposure.Among the large amount of data collected so far, divergent results have been obtained, but this could be a consequence of several limitations intrinsic in each study. For example, some studies are limited by the use of DNA from different tissues or the use of different surrogate methods of quantifying DNA methylation. Other studies are constrained by the method of quantification of the exposure to cigarette smoke. Some studies determine DNMT1, DNMT3A, and DNMT3B). In 2009, two seminal papers described the existence of a new epigenetic modification, 5-hydroxymethylcytosine (5hmC), that was present in high levels in neurons and embryonic stem (ES) cells and arises from the oxidation of 5mC by a group of Fe2+ and 2-oxoglutarate-dependent dioxygenases belonging to the mammalian TET family, namely TET1, 2 and 3 [The most studied and best-characterized epigenetic mark is 5-methylcytosine (5mC), regulated by the action of various DNA methyltranferases  control gene expression by base-pairing to the 3’-untranslated region of the target messenger RNA, which results in problems translating the messenger RNA (mRNA) into a protein molecule, or in degradation of the target mRNA. It is also known that a single miRNA has the capability of regulating a number of genes , some ofMaccani and colleagues conducted a study on placental cell lines, to examine the effects of two components of tobacco smoke (nicotine and benzo(a)pyrene), on four candidate miRNAs expressed in the placenta. They showed downregulation of three distinct miRNAs miR-16, miR-21 and miR-146a, involved in cell cycle regulation, growth, immunomodulation and development in the placenta . In a foOnly a small number of studies have examined the associations between prenatal smoke exposure and histone modifications. A recent study, however, found an association between cigarette smoke exposure during spermatogenesis with abnormal histone-to-protamine transition in human sperm . The excThe consequences of these alterations in the histone/protamine exchange ratio and the SP1/SP2 ratio can lead to infertility and decreased quality of sperm. They also can lead to increased DNA damage that can be transmitted to the fetus ,72.in utero smoke exposure [As this review highlights, the research community has placed particular focus on investigating the influence of maternal cigarette smoking during pregnancy on DNA methylation patterns. Meanwhile, work on the effects of maternal cigarette smoking on miRNA expression, histone modifications or imprinting during pregnancy has been much less comprehensive. These are important future considerations for investigation in the context of exposure .Variation in mtDNA content is another effect deriving from the exposure to cigarette smoking . Due to The proper functioning of mitochondrial DNA depends on its integrity and the number of copies, suggesting that even slight alterations in mtDNA sequences could have profound effects. For example, mtDNA mutations in respiratory chain genes lead to an increase in ROS.The malfunction of the mitochondria due to mitochondrial DNA mutations and reduced functionality of the respiratory chain lead to a change in the number of copies of mtDNA and to an increase of mitochondrial mass. The increase of the number of copies of mtDNA, according to some researchers, is considered a compensatory mechanism in respiratory function ,78,79. TA few studies have examined the relationship between maternal smoking and mitochondrial functionality in humans as well as in animals, and the results are controversial. In humans, studies have reported dose-dependent associations between mtDNA content and smoking in saliva and lung tissue ,80.Regarding the prenatal effects, Bouhours-Nouet and colleagues  measuredin utero smoke exposure, IUGR and the increase in mtDNA copy number, suggests that cigarette smoke can also affect fetal growth by altering mitochondrial function. Some suggest that the increase in the number of copies of mtDNA is a compensatory mechanism to offset the reduction of mitochondrial functionality [Moreover, maternal smoking during pregnancy is associated with adverse outcomes, among which intrauterine growth retardation (IUGR) is the most frequent complication ,85,86. Tionality ,87.in utero or neonatal second hand smoke, Fotterman and colleagues noticed an increase in mtDNA copy number in aortic tissue [Maraca mulata showed a decrease in mtDNA copy number in aortic tissue after perinatal exposure to ETS accompanied by a significant increase in oxidative stress, mitochondrial damage and dysfunction [In mice exposed to c tissue . Anotherfunction .DNMT1, DNMT3BA and DNMT3B) [TET1, TET2, and TET3) [Mitochondrial DNA alterations have been proven as an important aspect of mammalian mitochondrial physiology and, similarly to nuclear DNA methylation, abnormal mitochondrial DNA could be the consequence of environmental stress . There a DNMT3B)  and TET nd TET3) .mtTFA) gene promoter, leading to subsequent mitochondrial respiratory complex dysfunction. They also demonstrated for the first time the therapeutic efficacy of treatment with demethylating agents in the reversion of these adverse effects [The role of environmental influences on mtDNA has been the focus of a limited number of studies, but some interesting data has already been collected, such as the discovery of higher methylation levels within the Phe-mtRNA and 12S rRNA coding regions in workers exposed to airborne pollutants ,93,94. A effects .DNMT1 in the mitochondrial matrix, bound to mtDNA and modifying transcription of the mitochondrial genome, there is no evidence in the literature regarding a hypothetical function of the TET family of methylcytosine oxygenases in the mitochondria. However, both epigenetic modifications (5mC and 5hmC) have been found in mtDNA. DNMTs and TETs proteins have also been identified in the mitochondria in DNA samples extracted from blood and cultured cells from both humans and mice. In addition, the majority of methylated cytosines in mitochondria are located outside of CpG nucleotides [In contrast with the recent characterization of leotides ,93 TogetExposure of maternal and paternal gametes to biological, chemical and physical agents before conception can have profound effects on the newborn. Unfortunately, the association of exposures prior to conception with genomic stability in the offspring is difficult to analyze in human populations, because it can not be separated from gestational exposures. Maternal environmental smoke exposure affects the health of the developing individual and increases the risk of congenital malformations . There aInterestingly, cigarette smoking has been directly linked also to numerical chromosome abnormalities in gametes, which finally determine serious infertility problems, such as pregnancy loss, or developmental disabilities and mental retardation in the progeny . Some stOther studies have analyzed the consequences of cigarette smoking on the vitality and health of human gametes. For example, sperm form smokers have a higher level of DNA strand breaks  and highLaubenthal and colleagues analyzed the genomic stability in the offspring exposed to pre-conceptional paternal smoking and gestational maternal smoking, to discriminate between gestational exposure and exposure prior to conception in 39 families . They shCigarette smoke metabolites may alter the genomic stability inducing DNA damages such as epigenetic modifications, which can be transmitted to the next generation. Youngson and Holliday demonstrated that cytosine methylation and chromatin structure patterns in spermatozoa were transmissible throughout subsequent generations ,112. MetGerm-line and post fertilization reprogramming are two extremely important and vulnerable phases for the development of the fetus . If reprEnvironmental factors, even in the absence of continued exposure, may affect DNA reprogramming, persist through next generations and compromise their health  definingTherefore, it can be assumed that environmental factors such as cigarette smoking may have the same effect, especially if we consider that cigarette smoking is one of the most powerful agents capable of modifying DNA methylation . Howeverin utero [Moreover, hypothetically, epigenetic and DNA damages in nuclear DNA can also influence the regulation of genes responsible for maintaining genome stability and alter regular DNA damage response and repair processes. This hypothesis is a possible explanation for the elevated strand breaks found in cord blood DNA from newborns exposed to cigarette smoke in utero ,125.Tobacco smoke ranks as an important public health problem whose adverse impacts have spread around the world. The concern increases if we consider the association between exposure to prenatal smoke and increased risk of spontaneous abortion, preterm delivery, respiratory disease, immune system difficulties such as asthma and allergies, placental complications and increased risk of illnesses, health problems, cancer and psychological outcomes later in development ,126,127.Most of the research about the health effects of cigarette smoke have focused on mutagenic and genotoxic effects on nuclear DNA, but more recently, epigenetic studies have shown that cigarette smoke can influence the parental, fetal, or placental epigenome, altering gene regulation mediated by DNA methylation, histone modifications and noncoding RNAs. Genetic and epigenetic effects of cigarette smoke could contribute together to compromise the health of the individual but, the relationships between them still needs to be better understood. However, the literature reports that cigarette smoking acts mainly on some target regions on DNA, such as genes of the chemical detoxification pathway.CYP1A1 is a gene coding for a Phase I enzyme and is often reported in the literature in relationship to cigarette smoke [CYP1A1 affect susceptibility to tobacco smoke [CYP1A1 site-specific CpG methylation and gene expression [CYP1A1 in particular, but also other genes involved in chemical detoxification pathway, may represent a specific target for tobacco smoke, which can modify its function in a genotoxic or epigenetic way.As explained before, the genotoxic potential of tobacco compounds require a metabolic activation by Phase I detoxification enzymes, while Phase II enzymes, catalyze the activation or deactivation of aromatic and heterocyclic amine carcinogens. te smoke ,45. In fco smoke . Furtherpression ,47. CYP1Importantly, cigarette smoke contains and generates various ROS, which can damage nuclear DNA but also mtDNA. Recently, many groups have noted variation in mtDNA copies number, in response to damage to mtDNA and the respiratory chain, associated to cigarettes smoke exposure ,128,129.Mitochondrial DNA is extremely vulnerable. Defects in the respiratory chain can lead to an increased production of ROS, increasing in turn the probability of nuclear DNA damage and increases in mtDNA content, a compensatory mechanism for the damages to the respiratory chain and the lack of energy production ,80. MoreIn conclusion, cigarette smoke leads to adverse effects on nuclear DNA and mtDNA, affecting the health of newborns by compromising their energy intake. Furthermore, since mtDNA is maternally inherited, it is conceivable that maternal exposure to cigarettes smoke prior to conception can lead to mtDNA alterations, which are subsequently transmitted to new generations. More investigations on humans will be necessary to clarify the influences of cigarette smoke on mtDNA and the interactions with genomic DNA alterations, opening another chapter in the understanding of cigarette smoke mechanisms of action.in utero and perinatal. Not much attention has been placed on the preconception period, mainly due to the difficulty of distinguishing pre-conceptional effects to those that occur in the two later periods. However, the literature reports chromosomal aberrations and DNA damages in parental gametes, in particular in sperm [in utero and early life periods.In addition to providing an overview on the mechanisms of action of cigarette smoke adverse effects on nuclear and mtDNA, this review wants to emphasize the importance of parental life-styles in the future health of their unborn children. As mentioned before, there are three sensitive periods during the very early stage of development where a disruption or modification of the environment can influence fetal and offspring development: preconception, in sperm ,108,109 in sperm ,23,25,27in sperm . More st


Here we reconstruct the evolutionary history of eumalacostracan  development with regard to early appendage morphogenesis, muscle and central nervous system development, and determine the heterochronic transformations involved in changes of ontogenetic mode.Within Malacostraca (Crustacea), direct development and development through diverse forms of larvae are found. Recent investigations suggest that larva-related developmental features have undergone heterochronic evolution in Malacostraca. In the light of current phylogenetic hypotheses, the free-swimming Parsimov-based genetic inference (PGi). Our results confirm previous suggestions that the event sequence of nauplius larva development is partly retained in embryogenesis of those species which do not develop such a larva. The ontogenetic mode involving a nauplius larva was likely replaced by direct development in the malacostracan stem lineage. Secondary evolution of the nauplius larva of Dendrobranchiata from this ancestral condition, involved only a very small number of heterochronies, despite the drastic change of life history. In the lineage leading to Peracarida, timing patterns of nauplius-related development were lost. Throughout eumalacostracan evolution, events related to epidermal and neural tissue development were clearly less affected by heterochrony than events related to muscle development.Timing of 33 developmental events from the different tissues was analyzed for six eumalacostracan species  and one outgroup, using a modified version of nauplius larva in Malacostraca.Weak integration between mesodermal and ectodermal development may have allowed timing in muscle formation to be altered independently of ectodermal development. We conclude that heterochrony in muscle development played a crucial role in evolutionary loss and secondary evolution of a The online version of this article (doi:10.1186/2041-9139-6-4) contains supplementary material, which is available to authorized users. Heterochrony—evolutionary change in timing of developmental events—is a central concept in understanding the diversity of animal form –3. InvesnaupliusParsimov- Larsson  would giThe following questions will be addressed :How did the last common ancestor (LCA) of Eumalacostraca develop and to what degree did it show larval developmental patterns?Which heterochronies were involved in evolution of the nauplius larva of Dendrobranchiata?Which changes of the developmental sequence caused the emergence of zoea-like larval forms?In which way were developmental sequences altered in the lineage leading to Peracarida?P. fallax forma virginalis is described in  because they are always observed to occur simultaneously.Myogenic events (8–25): First appearance of the muscle precursors described in , [A2], [Md] , the first muscle precursors become detectable in the second antenna segment [a2-l]. In Si EN17 (17hpf), anlagen of the protocerebral, deutocerebral, tritocerebral, and mandible ganglia appear [NGA] . Si EN23 (23hpf) is characterized by distinct nerve fiber bundles forming the circumesophageal ring, the presence of the nauplius eye primordium [NEA] and the posterior pioneer neurons [PPN]. Si N1 (nauplius stage 1) hatches from the egg membrane [HAT]. In Si N3 (nauplius stage 3), appendage buds of the fist maxilla, second maxilla, first thoracopod, and second thoracopod appear [Mx1], [Mx2], [T1]  shows anlagen of postnaupliar ganglia  shows ganglion anlagen in the first and second thoracopod segment [t1-g], . In Pf III, ganglion anlagen of the most posterior trunk segment are still lacking, but continuous longitudinal neurite bundles are already present  neuromere showing connectives and lateral nerves with advanced differentiation  of the amphipod Orchestia cavimana. Appendage buds in the sixth pleon segment are seen in semaphoront Ph E5 [P6]. In Ph I, the first muscle precursors appear: [st], [lmp-t1]. Ganglion anlagen of the ventral nerve cord are visible down to the third pleon segment  is characterized by first appearance of stomodeal muscle precursors [st]  shows first appearance of naupliar appendage buds, naupliar ganglia, as well as the naupliar appendage muscle primordia [A1], [A2], [Md], [NGA], [a1-m], [a1-l], [a2-m], [a2-]l, [md-m], [md-l]. We specify a third embryonic semaphoront Af EN3, to which we assign the appearance of the nauplius eye [NEA], according to published data , they still occur near the beginning of the sequence. This position is not altered on the branches leading to Caridoida and Decapoda.PGi analysis shows several heterochronic changes in the eumalacostracan stem lineage. [e Figure . The egglmp-p6] are shifted late while formation of the second antenna muscle precursor [a2-m] is shifted early. The egg nauplius phase of the reconstructed caridoid LCA consists only of naupliar appendage bud formation . The first maxilla bud [Mx1] is still formed after the second maxilla bud [Mx2] while postnaupliar ganglia maintain their anteroposterior progression. Formation of appendage buds [P6], ganglion anlagen [p6-g], and longitudinal muscle precursors [lmp-p6] are now concentrated near the end of the sequence.In the caridoid stem lineage formation of the first maxilla bud [Mx1], the sixth pleopod bud [p6] and the longitudinal muscle precursor [md-m] and [a2-m]  was a ‘raconata  (Pancrusricto in ) 17]. O. Osensu Nebalia bipes has been described  allows trunk functionality before the posterior trunk segments are fully developed, as is the case in zoea-like larvae. Further common features of zoea-like larva development in terms of developmental timing are rapid formation of appendages in an anterior set of postnaupliar segments , late formation of pleonal segments [P6], [P6-g], [lmp-p6] (with the exception of the stomatopod pseudozoea), and late offset of segment formation in the germ band [FS]. Event data for the second thoracic to the fifth pleonal segment was not analyzed because the data could not be acquired for a sufficient amount of species and semaphoronts.In Malacostraca, both ectodermal and mesodermal tissues, from the second maxillary to the sixth pleonal segment, are formed sequentially by proliferation of stem-like cells (ectoteloblasts and mesoteloblasts) in the posterior growth zone , 107. Thke larva , 73. lmpzoea-like larva likely evolved independently in the lineages leading to Stomatopoda and Decapoda and that the eumalacostracan last common ancestor developed directly. The posterior longitudinal muscle primordium [lmp-post] which we consider a necessary feature for zoea-like larval motility was acquired twice independently in the lineages leading to Stomatopoda and Decapoda and was not part of the eumalacostracan ground pattern. The appendage bud and longitudinal muscle precursor of the sixth pleonal segment are formed early in the sequence reconstructed for the eumalacostracan last common ancestor, suggesting that functionality of the trunk did not precede differentiation in the posterior pleon segments, and consequently that direct development rather than a zoea-like larva constituted the developmental mode. We point out that the conclusions on this early node should be treated with caution because variation in the data set is extensive and also because we cannot rule out the possibility of bias due to the strong representation of direct development in the analysis.The scenario reconstructed by PGi suggests that a zoea-like larva as hatching stages appears well supported. In the developmental sequence of the decapod ground pattern, appendage bud formation and formation of the longitudinal muscle precursor in the sixth pleonal segment occur close to the end of the sequence. Offset of segment formation also occurs late while the anterior postnaupliar appendage buds are formed simultaneously and just after the egg nauplius phase. Also all anterior longitudinal muscle precursors and lmp-post are formed simultaneously. The same is true for the extrinsic appendage muscle precursors of the anterior postnaupliar segments. In the lineage leading to Dendrobranchiata, timing of postnaupliar events relevant to zoea-like larva formation remains nearly unchanged (with the exception of [t1-g]). The evolution of the novel developmental mode of Dendrobranchiata, involving the novel larval stages metanauplius, protozoea and mysis stages, from an ancestral condition with a more extensive embryonic period did not depend on changes in developmental timing of the analyzed morphogenetic events. It is the predisplacement of the hatching event that makes the actual difference between larval and embryonic development, while the sequence in which appendage buds, ganglion anlagen, and muscle precursor are generated remains largely unchanged. Certainly, acceleration of differentiation processes which follow the formation of appendage, ganglion, or muscle anlagen in the segments of a viable free-swimming larva must be assumed for the evolution of the dendrobranchiate ontogenetic mode. In the lineage leading to Pleocyemata, [FS] and [lmp-p6] are shifted to earlier positions. Both changes point toward loss of zoea-like larva formation, but the majority of relevant events is still in place. Both pleocyemate representatives  develop directly and lack larval stages. Therefore, we consider early placement of [FS] and [lmp-p6] as bias toward direct development in our data set, not necessarily as part of the pleocyemate ground pattern.For the last common ancestor of Decapoda, a Nauplius or zoea-like larvae are not found in peracarids. Changes to the developmental sequence in the peracarid stem lineage comprise loss of the nauplius eye and the posterior pioneer neurons, as well as late shift of several naupliar and early shift of postnaupliar events. In the ancestral peracaridan developmental sequence, naupliar and anterior postnaupliar appendage buds are formed rapidly at the beginning of the sequence, while naupliar ganglia are formed late. Offset of segment formation and formation of the posteriormost pleonal appendage bud occur early while the majority of muscle precursors are formed late. These properties of the developmental sequence do not resemble zoea-like developmental timing patterns. The condition in the caridoid ground pattern is difficult to interpret in terms of developmental mode. The late position of pleonal events suggests zoea-like larva formation but the posterior longitudinal muscle primordium [lmp-post] is not formed in the sequence. The question whether direct development might be plesiomorphic for Peracarida can therefore not be answered at this point.Peracaridan development is derived in many respects relative to the malacostracan ground pattern, because of the advanced mode of brood care that is autapomorphic to this group , 108–110P. hawaiensis (Amphipoda) the adaptation to efficient formation of juvenile body structure is intensified. Here the events of the six anterior segments appear in an order corresponding to the tissues they belong to: appendage buds, followed by ganglion anlagen, followed by muscle precursors. Together with the early offset of segment formation, this suggests a strong acceleration of morphogenesis in this lineage, which resulted in more rapid anteroposterior progression of segment formation and an earlier onset of tissue differentiation.Within Peracarida, most likely development was consistently adapted to efficient formation of juvenile body morphology after the advent of the new developmental mode, which was constrained by the specialized mode of maternal brood care. In nauplioid’ ’ (the event is coded from the given literature source), ‘scientists name, personal communication’ . (XLSX 29 KB)Additional file 1: Nexus file for analysis of heterochrony. Nexus file containing the rank-matrix and the tree topology. The data is formatted for use with PGi under R’s ape package. A rank value is coded for every event shown in Table Additional file 2: PGi superconsensus tree. Superconsensus tree generated by PGi-analysis from pseudoconsensus trees of three independent runs with algorithm parameters set to 100 cycles of selection per node, 200 sequences per cycle of selection, a maximum of 100 ancestral developmental sequences to be retained at each node and ‘semi-exhaustive’ pseudoconsensus setting with a limit of 3,000 evaluated solutions of equal score. Developmental sequences in the superconsensus tree are shown using only the event numbers (shown in Table Additional file 3:


Cardiac resynchronisation therapy (CRT) is an accepted treatment for heart failure patients with depressed left ventricular (LV) function and dyssynchrony. However, despite better clinical outcome and improved cardiac function after CRT in the majority of eligible heart failure patients, a large proportion of implanted patients do not seem to benefit clinically from this therapy. In this review we consider whether genetic factors may play a role in modulating response to CRT and summarise the few genetic studies that have investigated the role of genetic variation in candidate genes. Patients with heart failure and decreased function often develop dyssynchronous contraction of the ventricles because of electric activation delay. This further decreases systolic function and chamber efficiency and contributes to morbidity and mortality . ArtificSignificant interest exists in the identification of factors that predict response to CRT, as these may contribute to tailoring of the treatment of heart failure in the individual patient. Several demographic, clinical and procedural variables that could be possible predictors of CRT success have been intensively investigated. Factors implicated in response to CRT include the aetiology of heart failure, gender, QRS duration and morphology, and myocardial scar burden , 6. As fThe genetic architecture of the CRT response phenotype is undoubtedly complex as demonstrated for other clinical cardiovascular traits such as heart failure , atrial NR3C2, encoding the mineralocorticoid receptor, was found to be higher in the group that did not display reverse remodelling according to the criteria used in the study. The same case-control set used in the above study was subsequently used to test an additional 5 candidate SNPs that had been previously associated with a variety of cardiovascular disease phenotypes [ATPIB1, rs5443-GNB3, and rs7325635-TNFSF11) appeared to be associated with success of CRT. Following pre-clinical observations made in dog [ADRB1-Arg389Gly, ADRB2-Arg16Gly and ADRB2-Gln27Glu). They conducted their study in a consecutive cohort of 101 heart failure patients who underwent CRT or CRT-D (70 patients) implantation and who were assessed at baseline and after a follow-up of one year. They demonstrated that carriership of the Glu27 allele of the ADRB2-Gln27Glu polymorphism was associated with a greater increase in LVEF, and in an additional analysis was associated with less cardiac death and appropriate ICD discharge. Interestingly, the Glu27 allele has also been shown to predict a positive reverse remodelling response to the β1/β2/α1-blocker carvedilol in heart failure patients [As far as we could determine, three studies have so far searched for genetic factors modulating CRT response. All three investigated the effect of common genetic variants in candidate genes. One study focused on 7 genetic variants at 5 genes from the renin-angiotensin-aldosterone system (RAAS) , a pathwenotypes . Of these in dog  and cline in dog , suggeste in dog  studied patients –25, and patients . It is iTwo recent studies investigated the role of microRNAs (miRNAs) in CRT response , 28. Mic2+ handling, β-adrenergic receptors, contractile proteins and myocardial natriuretic peptide [Data on mechanisms underlying dyssynchrony and differential response to CRT are currently scarce and there would be a clear benefit in this regard if genetic studies were to identify possible molecular players. Ion channel or gap junction dysfunction that promotes conduction delay is clearly a highly likely candidate for the development of dyssynchrony. Similarly, structural and contractile proteins are likely involved in the development of heart failure following dyssynchrony. It is plausible that these same pathways may determine response. Gene expression studies in serial biopsies obtained pre- and post-CRT in patients have identified changes in the expression of genes encoding components of Ca peptide –33. In a peptide . Other e peptide , 35). El peptide . Yet, fuWhile the identification of genetic factors that modulate response to CRT is a relevant quest, since it may contribute to the ability to distinguish responders from non-responders, their identification is highly challenging considering the complexity of the phenotype and the likely complex underlying genetic architecture. In practical terms a number of obstacles, therefore, need to be overcome to enable such genetic studies. These relate primarily to the necessity of large biobanks of deeply and uniformly phenotyped patients. Once such biobanks become available, we can then dissect the possible role of genetic factors through the investigation of candidate pathways (as directed by mechanistic studies in model systems and knowledge on intermediate phenotypes), or in an unbiased genome-wide fashion using current genomic technologies Fig. . Such apThe authors are supported by research grants from the Center for Translational Molecular Medicine  and from the Netherlands CardioVascular Research Initiative  (PREDICT project).None declared.


A remarkable rise in life expectancy and the declining prevalence of certain causes of late-life mortality allow dementia to become a common occurrence with age. Alzheimer's disease (AD) is the most prevalent form of dementia characterized clinically by progressive loss of memory, and pathologically by the presence of neuritic plaques and neurofibrillary tangles . Many sc+/− triple mutant mice were used. Similar to AD patients, ASM activity was elevated in plasma, fibroblast, and brain  of APP/PS1 mice. This activity was decreased in APP/PS1/+/−ASM mice to levels within the normal range or lower. Strikingly, AD pathology including Aβ deposition and memory dysfunction was significantly reduced in APP/PS1/ASM+/− mice [In a recent study published in Journal of Experimental Medicine, we reported that increased ASM activity in AD results in defective autophagy due to disruption of lysosomal biogenesis . ASM act+/− mice .+/−ASM mice revealed a reduced these vesicles, indicating that elevated ASM activity in AD causes autophagy dysfunction and that partial genetic inhibition of ASM leads to restoration of autophagy process. Similarly, the inhibitory effect of ASM in pathogenesis of AD also has been confirmed in amitriptyline-hydrochloride  treated APP/PS1 mice [There is growing evidence to suggest that dysfunction of the proteolytic degradation system could affect AD pathogenesis and lead to enhanced Aβ deposition. Autophagy, a process to destroy misfolded proteins or breakdown faulty components and then recycle, is known to be markedly impaired in AD . We founPS1 mice .To examine the direct relationship of ASM and autophagic dysfunction, we analyzed recombinant ASM treated human neuron and fibroblast. We found that increased lysosomal ASM induces autophagosome accumulation in a concentration dependent manner. We also identified that ASM causes a defect of autophagic degradation, but not induction . Recentlin vitro and in vivo results are paralleled by similar alterations in patients, human AD fibroblasts (PS1-FAD and apoE4) and induced pluripotent stem cells-derived neurons with PS1 mutation were analyzed. We found that abnormal autophagy observed in AD mice by ASM also occurs in AD patient-derived fibroblast and neuron, and restoration of ASM back to normal levels is able to ameliorate autophagic dysfunction by restoring lysosomal biogenesis in AD patient cells.To explore whether the observed effects of ASM The combination of the vast knowledge of lipids and their biophysical and biochemical properties is translated into the development of new therapeutic and diagnostic modalities for diverse diseases. Similarly, the field of sphingolipids becomes very active and popular as a result of the understanding of sphingolipids' role in disease environment. This dramatic increase in the understanding of the various functions of sphingolipids leads to the development of several novel drugs, the first of which, a sphingosine-1-phosphate agonist for multiple sclerosis, was recently approved by the Food and Drug Administration . The res


Parthenogenetic activation of human oocytes obtained from infertility treatments has gained new interest in recent years as an alternative approach to create embryos with no reproductive purpose for research in areas such as assisted reproduction technologies itself, somatic cell, and nuclear transfer experiments and for derivation of clinical grade pluripotent embryonic stem cells for regenerative medicine. Different activating methods have been tested on human and nonhuman oocytes, with varying degrees of success in terms of parthenote generation rates, embryo development stem cell derivation rates. Success in achieving a standardized artificial activation methodology for human oocytes and the subsequent potential therapeutic gain obtained from these embryos depends mainly on the availability of gametes donated from infertility treatments. This review will focus on the creation of parthenotes from clinically unusable oocytes for derivation and establishment of human parthenogenetic stem cell lines and their potential applications in regenerative medicine. Aspidoscelis uniparens) [Parthenogenesis is a reproduction strategy common in some jawed vertebrate species such as the whiptail lizard (iparens) , in whiciparens) . No furtiparens) . On the iparens) .Why then do we study parthenogenesis in the scope of assisted reproduction technologies (ART) if no human baby can be generated from this reproductive maneuver?Renewed interest in parthenogenetic activation of mammalian oocytes has come about for scientific, medical, and economic reasons. First of all, experimental work involving parthenogenetic embryos circumvents the ethical and legal problems concerning the use of human embryos generated for reproductive purposes. Parthenotes may be employed in different research areas related to ART, studies on human pluripotent stem cells or basic science uncovering regulatory mechanisms that command human embryonic development and cloning experiments using somatic cell or nuclear transfer in mammalian oocytes.Last but not least, the creation of clinical grade parthenogenetic human embryonic stem cell (hpESC) lines have the potential to benefit a considerable large number of patients, when used in cell therapies, with a reduced risk of transplant rejection. However, to this end, efforts should be directed to clinicians to participate in research projects. The establishment of parthenogenetic clinical grade human embryonic stem cell lines for use in cell or tissue therapies will only be possible, if research laboratories obtain enough biological material from ART centers to evaluate activation strategies, derivation, and culture media adequate for future therapeutic use of phESC lines. Furthermore, well trained committed staff, with good practice and knowledge of basic embryology, is paramount for the conscious and successful use of spare oocytes and embryos obtained from assisted reproduction centers.This review will explore techniques of parthenogenetic activation, derivation, and cell and tissue differentiation achieved in animal models, which reinforce the therapeutic potential of these pluripotent embryonic stem cells, despite concerns related to their putative genetic instability, an inherent feature due to their monoparental origin .ART In Vitro Fertilization and Intracytoplasmic Sperm Injection Techniques. Among the several treatments regarded as ART, there are two insemination methods employed for the generation of human embryos in the laboratory, the in vitro fertilization (IVF) and the intracytoplasmic sperm injection (ICSI) (n (ICSI) . cumulus cells, interact with the zona pellucida proteins, and eventually attach and fuse with the oocyte plasma membrane.When performing IVF, oocytes and sperm are mixed together and left to undergo the fertilization process in a culture system that mimics the fallopian tubes environment. Thus, IVF may be considered an assisted reproduction technology that mimics “natural” fertilization phenomena . With IVICSI was developed to overcome situations where sperm count, motility, or morphology is compromised. By using this insemination technology, the embryologist may choose the “best looking” spermatozoon to inseminate the oocyte, even if it is in very low numbers or presents limited motility in the ejaculate.ICSI is performed with the aid of a pair of glass pipettes adjusted to an inverted microscope, where the embryologist holds the oocyte with one pipette and injects the chosen spermatozoon with the second pipette straight into the ooplasm.2+ rises and meiosis resumption. Intracellular Ca2+ oscillations will inhibit the action of the metaphase promoting factor (MPF) and the cytostatic factor (CSF) and lead to metaphase/anaphase transition, segregation of sister chromatids, and extrusion of second polar body. The importance of these Ca2+ transients for oocyte activation was shown by the prevention of the intracellular elevation of Ca2+ after sperm penetration by preloading the oocytes with the Ca2+ chelator BAPTA 1-AM [2+ increase, activation and subsequent embryo development failed to occur. In mammals, intracellular Ca2+ transients are triggered by a putative sperm factor, the testis specific phospholipase C-zeta (plc-zeta), or the newly described postacrosomal sheath WW domain-binding protein PAWP released by the sperm during the normal fertilization process [2+ oscillations, its presence is paramount for successful oocyte activation and embryonic development. The activating agents used for parthenogenetic activation may mimic the sperm bound stimuli to trigger Ca2+ release from the endoplasmic reticulum, exit from meiotic arrest, and start embryonic development.Arrested nonfertilized metaphase II (MII) oocytes will remain at this stage until a stimulus, which may come from the fertilizing spermatozoon or from an artificial agent , triggerPTA 1-AM . In the  process . Regardl2+ [2+ oscillations normally observed during fertilization. However, the single rise in intracellular Ca2+ that is produced by agents such as Ca2+ ionophore, ionomycin, or ethanol is adequate to trigger the meiotic resumption and cortical granule release. Specific oocyte activation protocols using strontium chloride (SrCl2) to induce continuous Ca2+ oscillations during exit from second meiosis and first embryonic mitosis seem to have a role in long-term embryonic events, such as the number of cells in the inner cell mass (ICM) and trophectoderm of the resulting blastocyst [A wide range of artificial stimuli have been employed as agents to trigger the activation process in oocytes of several animal species. All the techniques which have been described for artificial activation can be divided into two major groups, depending on the nature of the activating stimulus, that is, chemical or physical agents. Except for Sr2+ , all othastocyst .2+ elevations, it seems that there is not a single method that activates oocytes from every species studied. Thus, parthenogenetic activation strategies have presented varying degrees of success, regarding activation rates and subsequent embryonic development according to the protocol employed and the species.Despite the fact that various methods have been investigated for parthenogenetically activating oocytes through intracytoplasmic Ca2, ethanol, Ca2+ ionophore, or ionomycin is followed by another chemical, for instance, 6-DMAP (broad protein synthesis inhibitor) or cytochalasin B or D (inhibitors of actin filaments polymerization), which blocks second polar body (PB2) extrusion. Thus, the resulting parthenote is a “pseudodiploid” heterozygous embryo containing the two sister chromatids of each maternal chromosome present in the MII oocyte.In the first instance , treatmeAlternative activation methodology may allow PB2 extrusion and the resulting parthenote containing a single copy of the sister chromatids undergoes spontaneous diploidization giving rise to a completely homozygous embryo .Last, the activating agent induces exit from MII and PB2 extrusion, without diploidization . The res2+-free medium supplemented with 10 mM SrCl2 for 2 hours [High oocyte activation and blastocyst formation rates were achieved, when mouse oocytes were exposed to Ca 2 hours , 9. Howe2+ salt ionomycin for 5 minutes, followed by 2 mM 6-dimethylaminopurine (6-DMAP) for 3.5 hours [On the other hand, bovine oocytes presented high activation rates and development to the blastocyst stage, when exposed to the Ca.5 hours . This la.5 hours .Human oocytes have been artificially activated mainly with the aim to generate blastocysts for the creation of parthenogenetic human embryonic stem cell (phESC) lines . Notably2+ ionomycin together with 6-DMAP [The first phESC lines were created using Cah 6-DMAP . In thath 6-DMAP  describeh 6-DMAP , the innThe detection of nonfertilized oocytes after insemination is not uncommon in ART treatments, both due to gamete immaturity and unknown factor that led to a failure to fertilize (FF). Most frequently, FF occurs after IVF. In this situation, the oocyte will remain arrested in MII. Rescue insemination by ICSI, few hours after detection of FF, has been proposed as a valid alternative to induce the creation of zygotes from the nonfertilized oocytes –19. TherOn the other hand, it has been shown that unwanted, FF human oocytes retain their developmental potential and may generate viable blastocysts by exposure to ionomycin followed by 6-DMAP and a protein synthesis inhibitor, cyclohexamide . The emb2 or Ca2+ ionophore on the activation rates of 3-day-old FF human oocytes and observed that the treatment with the ionophore was more efficient in activating aged unfertilized oocytes and embryo development to the blastocyst stage. However, these reports on the activation of FF oocytes do not reflect the real parthenogenetic embryonic development potential of the human oocyte as the studies mentioned earlier [A recent study  compared earlier , 14, 15,2+ releasing agents, SrCl2, and ionomycin have been used aiming at activating oocytes during fertilization. In a series of nine cases of couples who have had total fertilization failure or very low fertilization rates in previous cycles, Kyono et al. [2. Fertilization rates increased from 21.7% in the previous cycles to 64.5% in the cycles using SrCl2 artificial activation. Six pregnancies were established, four of which went to term. Five healthy children were born and their 1-year followup did not show any physical or neurological abnormality. In another study, the same group of authors employed the Ca2+ ionophore to activate zygotes generated by ICSI using globozoospermic sperm [2+ ionophore to activate the oocytes during fertilization and induce embryo development and gestation in cases of previous FF of human oocytes was also reported with relatively good success rates by other groups [As an adjunct procedure in ART cycles, two intracellular Cao et al.  exposed ic sperm . The altr groups –27.2 or Ca2+ ionophore may have induced the release and oscillations of intracellular Ca2+ needed for oocyte exit from the meiotic arrest and embryonic development [The exposure to SrClelopment , 9. Howe in vitro maturation and ICSI to produce embryos for reproductive purposes. Resulting implantation and pregnancy rates are generally poor and there are controversies on whether to use these immature oocytes and embryos for reproductive purposes [ in vitro mature GV or MI oocytes obtained from stimulated ovaries may yield MII oocytes and potentially parthenotes, under suitable IVM conditions. Liu et al. [In classical, stimulated ART cycles, immature germinal vesicle (GV) or metaphase I (MI) oocytes are commonly collected together with the mature MII. The immature oocytes may be submitted topurposes . Howeveru et al.  have shoOn the other hand, IVM protocols to promote maturation of oocytes collected from unstimulated ovaries exist and they are employed in several fertility centers around the world, as an alternative to classical, stimulated cycles –32. The The uniparental origin of parthenogenetic SCs (pESCs) confers its great advantage of producing immunocompatible pESCs , an adeqMany aspects related to the biology of these cell lines, however, still need to be elucidated. Probably one of the major concerns of using pESCs cell for regenerative medicine is the inability of parthenote mammal embryos to support full-term development. It is known already that the accurate establishment of epigenetic regulation and maintenance of genomic imprints during embryogenesis are essential for normal embryonic/placental development . The genIn parthenotes, the lack of sperm alleles leads to the transcription of both alleles of maternally expressed imprinted genes and the absence of paternally expressed imprinted genes, possibly causing the overexpression of imprinted genes often associated developmental abnormalities , 43. IndAlthough parthenogenetic embryos retain aberrant imprints and consequently cannot develop to term, established parthenogenetic ESCs were reported morphologically indistinguishable from ES derived from fertilized embryos and may also present normal gene expression or even correction genomic imprinting in chimeras, when pESCs were used in tissue contribution . Hence, Whereas bone marrow transplantation is an example of stem cell therapy that is in clinical use worldwide, the therapeutic use of pluripotent stem cells, such as human embryonic stem cells (hESCs) or phESCs, is still in its infancy. First clinical trials in the US using hESCs involved specific conditions such as macular degeneration . The widTaken together, these observations indicate that pESC lines have the potential to be used in regenerative medicine, not only for autologous transplantation into oocyte donors, but also for creation of a bank of histocompatible cell lines. However, the possibility of an interplay of epigenetic reprogramming on a monoparental background, together with the lack of paternal imprinted genes expression, cast caution on hpESCs therapeutic application. Thus, additional research is needed to ensure the safety of using parthenogenetically derived ESC lines in regenerative medicine.Despite all the advances in infertility treatments since the birth of Louse Brown in 1978, one challenging situation that remains to be overcome by ART is the patients with complete absence of gametes or gonads, who cannot have their genetically related child. Presently, gamete donation is the available alternative for these patients to conceive their children. Unfortunately, a worldwide shortage of altruistic gamete donors has exposed an increasing transaction of donor sperm and oocytes across borders . This un in vitro, but fundamental limitations of chromosome segregation and imprinting relegated this methodology to the realm of the “fantastic” [Tesarik  proposedntastic” . Howeverntastic” .In another elegant study by Duggal et al. , the aut in vitro differentiation of hpESCs has already provided completely different cell types [These findings represent important steps towards the development of novel therapies in cases where patients are entirely devoid of viable gametes. Considering thatll types , 58, the in vitro differentiation of human parthenogenetically derived clinical grade stem cells shall produce a wide range of cell types, which will eventually have many clinical applications, including ART procedures.It is clear from the information presented here that parthenogenetic activation of human oocytes rescued from infertility treatments results in embryos which are comparable to their biparental counterparts. The use of human parthenogenetic embryos as a source of cells for the generation of pluripotent stem cell lines is feasible and should be given more relevance in the present ESC research scenario. Considering the large number of oocytes discarded in infertility treatments , the in


ATG8e belongs to a core carbon signaling response shared by Arabidopsis accessions, and the transcription of Arabidopsis ATG7 is tightly co-regulated with genes functioning in chlorophyll degradation and leaf senescence. In the reproductive phase, autophagy is essential for bulk degradation of leaf proteins, thus contributes to nitrogen use efficiency (NUE) both under normal and low-nitrogen conditions.Carbon and nitrogen are essential components for plant growth. Although models of plant carbon and nitrogen metabolisms have long been established, certain gaps remain unfilled, such as how plants are able to maintain a flexible nocturnal starch turnover capacity over various light cycles, or how nitrogen remobilization is achieved during the reproductive growth stage. Recent advances in plant autophagy have shed light on such questions. Not only does autophagy contribute to starch degradation at night, but it participates in the degradation of chloroplast proteins and even chloroplasts after prolonged carbon starvation, thus help maintain the free amino acid pool and provide substrate for respiration. The induction of autophagy under these conditions may involve transcriptional regulation. Large-scale transcriptome analyses revealed that  Eukaryotic cells carry out autophagy to clean up the house and keep fit , 2010. Tin vivo, genetic and physiological studies . For instance, ATG7, encoding the E1-like activating enzyme for both ATG8 and ATG12 conjugation, is surrounded by key regulators and marker genes of leaf senescence such as MYB2, AtNAP, SAG12, and NYE1 . According to the guilty by association rule, ATG7 is likely a hub during plant senescence, when carbon is used for leaf energy and nitrogen gets remobilized , Arabidopsis mesophyll cells accumulate RuBisCO-containing bodies (RCBs) and structures containing pieces of chloroplasts in the lytic vacuole, whereas in atg4a atg4b-1 double mutants neither can be seen , represented by the nitrogen harvest index (NHI): Harvest index (HI) ratio, is lower than that of the wild-type both at the nitrogen-rich condition (+N) and over nitrogen limitation (-N). The lower NUE was shown to be independent of seed productivity (atg mutants is due to a defect of nitrogen remobilization leading to the accumulation of undigested soluble proteins in their leaves (Arabidopsis lines carrying 35S:GmATG8c, we found that the transgenic lines with higher levels of autophagy have comparable nitrogen concentrations to the wild-type at –N condition, yet maintain a higher biomass at both +N and –N conditions, and enter the reproductive phase earlier to produce more branches and more siliques at +N condition (uctivity . They alr leaves . Similarondition . Upon seondition . These rATG genes? Construction of higher-order gene regulatory networks will certainly be useful. Can the newly identified role of autophagy in starch degradation be integrated into the classic model of nocturnal starch turnover? The core machinery genes are generally controlled by the circadian rhythm; however, are they directly linked to the yet unidentified clock signals? Both mathematical modeling and well-planned screening may help answer these questions. Finally, the interaction between carbon and nitrogen has always been a vital topic in plant metabolism and signaling, and autophagy now appears to have a leading role (So far, studies have elucidated many basic molecular mechanisms and physiological and pathological consequences of autophagy in plants. The relationships between autophagy and plant carbon and nitrogen metabolism have started to be revealed. It can be expected that in the coming years, more interesting and fundamental researches will emerge to solve more existing problems in plant cell biology and plant metabolism. For instance, is there a common set of transcription regulators for the induction of plant ing role . The detThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


MLP is a member of the cysteine and glycine rich protein (CSRP) family that is composed by CSRP1, CSRP2 and CSRP3 or MLP. CSPRs, in turn, belong to the larger family of LIM domain proteins.2 MLP is expressed in striated muscle at the level of sarcomere, the intercalated disk and the costamere where it interacts with telethonin, α-actinin, cofilin-2 and calcineurin among others and contains two LIM domains that are zinc finger domains composed of ~55 aminoacids with 8 highly conserved residues, mostly cysteine and histidine that are located at defined intervals.2 Through the LIM domain, MLP can modulate protein interaction and formation of macromolecular structures. Another peculiarity of MLP is that it can shuttle from the cytosol to the nucleus where it serves as a scaffold protein to adapt transcription factors to their DNA-binding sequences.3 In fact, MLP is involved myocyte differentiation by activating transcription factors such as MyoD, myogenin, etc.4 To this effect, mutations in the N-terminal region of MLP have been linked to the development of dilated cardiomyopathy (DCM) or hypertrophic cardiomyopathy (HCM). Moreover, MLP protein levels or intracellular localization are altered in skeletal myopathies, such as fascioscapulohumeral muscular dystrophy, nemaline myopathy and limb girdle muscular dystrophy type 2B.6 Therefore, the current dogma foresee an important role for MLPin the maintenance of muscle cytoarchitecture as well as in the differentiation of muscle cells. However, the cellular and molecular mechanism(s) through which MLP carries out its function is (are) still obscure.The ongoing molecular investigation of the basic contractile units of the sarcomere in striated muscles have revealed a highly complex system composed of a large number of proteins each one with a specific task.7 In particular, we have used MLP silenced myoblasts to show an impairment of autophagy that, in turn, results in decreasedmyotubes formation and decreased resistance to cell death. However, the question was: how can MLP control autophagy? We provided evidences that MLP interacts and associates with LC3, a widely used autophagy marker with a crucial role during autophagosome formation. Interestingly both MLP and LC3 are microtubule associated proteins. Therefore, besides controlling muscle cell differentiation by modulating the expression of MyoD1 and myogenin mRNA, we also show that cytosolic MLP is important for the correct assembly and flux of the autophagic machinery. In fact, MLP silencing not only decreased LC3-II accumulation but also resulted in impaired degradation of long-lived proteins. Interestingly, morphological analysis of myotubes lacking MLP expression by transmission electron microscopy revealed the accumulation of damaged mitochondria and multi-membranous structures.Of note, the same multi-membranous structures with sarcomere disorganization have been described in skeletal muscles from Atg7–/– and Atg5−/− mice through the accumulation of damaged organelles and material. The final phenotype of these mice is characterized by muscle atrophy and features of myopathy.8 In fact, increasing evidences suggest that autophagy has an important physiological role during development, differentiation and muscle mass maintenance.9Using a myoblast cell line we have recently demonstrated that MLP uses macroautophagy to control differentiation of myobalsts into myotubes as well as survival of myotubes in the presence of a cell death stimulus.10 Our results show that reduced MLP expression with reduced autophagy, increases sensitivity of myotubes to apoptogens and cell death. This suggests that autophagy may represent a mechanism through which muscle cells can recover from a stress or a sub-lethal damage that do not kill the cell but requires extensive remodeling of internal structures.Autophagy has also been linked to another physiological pathway such as apoptosis. Like autophagy, apoptosis has a fundamental role during development and differentiation. In fact, autophagy and apoptosis uses common proteins and it is widely accepted that increased autophagy represent a protective mechanism from apoptotic cell death. Such knowledge derives from oncology in which cancer cells increases resistance to chemotherapy-induced apoptosis by increasing autophagic flux and removal of damaged organelles and proteins.de novo synthesis of structural genes, cytosolic MLP regulates autophagy thereby controlling recycling of organelles and proteins and providing building blocks for muscle cell cytoarchitectural remodeling.Our results shed new light on the MLP and on its central role for muscle cell differentiation and remodeling. As shown in In conclusion, even if mutations in MLP have been linked to DCM and HCM and deregulation of MLP to skeletal muscle diseases, new questions arise. For example: which are the pathogenic mechanisms activated by the lack or inactivation of MLP? Can MLP be used as a therapeutic target? The identification of the mechanism(s) of action of MLP may drive the development of new strategies for increasing skeletal muscle cell survival and remodeling.


Ovarian cancer (OC) mostly arises sporadically, but a fraction of cases are associated with mutations in BRCA1 and BRCA2 genes. The presence of a BRCA mutation in OC patients has been suggested as a prognostic and predictive factor. In addition, the identification of asymptomatic carriers of such mutations offers an unprecedented opportunity for OC prevention. This review is aimed at exploring the current knowledge on epidemiological and molecular aspects of BRCA-associated OC predisposition, on pathology and clinical behavior of OC occurring in BRCA mutation carriers, and on the available options for managing asymptomatic carriers. BRCA1 and BRCA2 are separate genes mapping on two different chromosomes . They have distinctive primary sequences nevertheless disruption of either BRCA gene leads to similar pathophysiological effects [BRCA1 and BRCA2 are considered tumor suppressor genes, since they are deputed to the maintenance of genomic stability and hence to the control of cell growth , 4. DSBsSeveral proteic interactors of BRCA1 and BRCA2 have been identified. RAD51 is responsible for repair mechanism of DSBs and is one of the most important players in HR; its functions are ultimately complemented by the proteins encoded by the two BRCA genes , 14. A nBRCA1 is a very large gene that generates several different transcripts. The full-length form is a 2843 amino acids p220) protein and a shorter (1399 amino acids) form, named BRCA1-IRIS, may have an oncogenic activity. BRCA2 is even larger, counting 3418 amino acids, but has fewer recognized motifs . BRCA1 a0 proteinDisease-associated mutations are scattered across the entire length of the BRCA1 and BRCA2 genes and usually result in a truncated protein. Deleterious missense mutations occur frequently in exons-encoding domains that interact with BRCA1-binding proteins, such as BARD1, BRIP1, and PALB2, which  are also breast and/or ovarian cancer susceptibility genes .The BRCA genes are routinely tested by Sanger sequencing of exons and exon-intron junctions. Whether a clearly pathogenic variant has not been identified, the multiplex ligation probe assay (MLPA) should be applied in order to exclude the presence of large BRCA1 deletions, involving one or more full-length exons. The MLPA assay is a rapid and robust method for copy number quantification and methylation status analysis of genomic sequence. It can be easily multiplexed and requires only a small amount of input DNA .The silencing of BRCA1 gene trough promoter hypermethylation may occur in sporadic breast and ovarian cancers . The anaIn a few years the next generation sequencing (NGS) technology, including different high-throughput sequencing systems, will likely replace Sanger sequencing as the technique of choice for genetic testing of BRCA genes because it undoubtedly offers advantages in terms of sensitivity, scale, and costs. However, the large number of false positive/negative insertions and deletions (indels) due to the high frequency of homopolymers in BRCA genes, which cause sequencing errors, has slowed down the usage of NGS for clinical genetic testing. The specificity of indels detection in NGS data is improving by the application of different filtering criteria for the variants calling .Based on the diagnostic criteria used in the United States to select patients to be tested, clearly pathogenic mutations in either BRCA1 or BRCA2 are found in 10% to 15% of hereditary breast and ovarian cancer families. Rare patients with mutations in both BRCA1 and BRCA2 genes were described , which hThe major challenge in the diagnostic testing of BRCA genes concerns the interpretation of unknown variants, the so-called “variants of uncertain significance” (VUS). The pathogenic variants can be nonsense mutations, small indels causing a frameshift, splicing site mutations that occur inside of the canonical splice sites, large deletions or known deleterious missense variants. VUS are alterations in the DNA sequence that have unknown effects on the protein function and disease risk; they usually are missense substitutions, splicing site mutations that occur outside of the canonical splice sites, small in-frame indels. Their frequency varies depending on the patient's ethnicity: in the United States, in individuals of European ancestry VUS account for approximately 5% of the alterations reported from BRCA genetic testing, but the estimate is as high as 20% among individuals of African ancestry . To guid in silico evaluation of the effect of each variant at the protein or mRNA level. Observational data, such as personal and family history, cosegregation of VUS with cancer phenotype in pedigrees, cooccurrence with other known pathogenic mutations, tumor immunohistochemistry, and histological grade, are summarized as likelihood ratios in favor of pathogenicity and used to update the prior probability [The method for VUS classification begins with a prior probability based on anbability , 33. Thebability .OC patients with a family history of breast or ovarian cancer present high probability of carrying a mutation in BRCA1 or BRCA2. The rate of mutations found in families with multiple cases of breast and ovarian cancer varies from 9% to 46%, depending on selection criteria and ethnicity –41. AmonIn addition, many efforts have focused on assessing the contribution of BRCA mutation to the total burden of OC. Early population-based studies reported BRCA1 mutation rates in OC patients varying from 1.9 to 7.2% –46.In a recent population-based cohort of 1001 Australian OC patients, 141 (14.1%) were found to carry a pathogenic mutation in BRCA1 (88 patients) or BRCA2 (53 patients), the proportion being 16.6% when only serous carcinomas where considered and increasing to 17.1% in patients diagnosed with high-grade serous cancers. 44% of mutation carriers failed to report any family history of breast or ovarian cancer . AnotherSeveral smaller studies have been carried out in different countries: in a Greek cohort of 592 patients with sporadic OC, a targeted screening of the commonest BRCA1 mutations detected 27 mutation carriers (4.6%) . In BelgThe vast majority of OC associated with germline BRCA mutations reported in the literature are high-grade and advanced-stage serous carcinomas . Most stAlso borderline BRCA1 carriers ovarian tumors are very rare , which rA large dataset, aimed at expanding the knowledge of OC pathology in these patients, has been recently reported by the Consortium of Investigators of Modifiers of BRCA1/2 (CIMBA) that represent the largest collaborative study of BRCA1 and BRCA2 mutation carriers, involving more than 37 groups from more than 20 countries . Tumor pMoreover, according to the recent update on the different pathogenesis and clinicopathologic features of ovarian low-grade and high-grade serous carcinomas, as well as to the fundamental molecular differences between both categories of tumors, the vast majority of BRCA-related hereditary ovarian tumors are high-grade serous carcinoma. Low-grade serous carcinoma and noninvasive micropapillary serous carcinoma do not seem to be related to germline mutations of BRCA .A high frequency of loss of heterozygosity (LOH) in or near the BRCA1 region has been reported in BRCA1-mutation positive OC . In addiThe term “BRCAness” has been used to describe the phenotypic characteristics that some sporadic OCs share with tumors found in the setting of BRCA germline mutations. The term also reflects that this common biologic behavior comes from molecular defects in the cellular machinery similar to those caused by BRCA mutation , 65. TheDespite being more aggressive than sporadic ovarian carcinomas, those arising in BRCA mutation carriers show higher susceptibility to platinum-salts and other DNA-damaging agents. Platinum-salts interfere with DNA cross-links creating double-strand breaks in the DNA helix, which cannot be repaired in BRCA due to HR deficiency. Studies demonstrated an improved long term-survival in women with OC treated with platinum-salts if compared to sporadic OC . IntrapeIn the last years the inhibition of the poly(ADP-ribose) polymerase enzyme (PARP) has emerged as a promising therapeutic approach in BRCA-associated OCs. PARP is a class of proteins which produces large branched chains of poly(ADP) ribose (PAR) from NAD+. They are involved in a number of cellular pathways including transcriptional regulation, DNA replication, and DNA damage repair , 80. Of The oral PARP inhibitor olaparib has shown to be well tolerated in phase I studies. A phase II study recruiting patients with BRCA1/2 mutations with recurrent OC demonstrated the efficacy of single agent olaparib with a median duration of response of 9.5 months and with a 66% of clinical benefit rate with the dose of 400 mg twice a day . At ASCOEvidence exists that OC patients carrying germline BRCA mutations have an improved prognosis in comparison to sporadic cases. A pooled analysis of 26 observational studies was recently carried out to explore survival differences between BRCA mutation carriers  and noncarriers (2666); BRCA mutation carriers showed a more favorable prognosis than noncarriers .A favourable prognosis for BRCA mutation carriers with OC was also supported by a recent retrospective study on 190 ethnically heterogeneous patients, 90 of whom were BRCA mutation carriers. A significantly longer overall survival was reported in mutation carriers compared with noncarriers , although no difference was observed in progression-free survival . ImproveBy comparing OC outcome in BRCA1 versus BRCA2 mutation carriers, Liu et al.  failed tAn Italian retrospective study found a survival advantage in BRCA2-positive versus BRCA1-positive OC patients, the median progression free survival being 45.46 months in the former and 27.2 in the latter . The betA study on the outcome of primary surgical cytoreduction in 69 BRCA mutated in comparison to 298 wild-type patients with high-grade serous ovarian carcinoma (FIGO stage IIIC-IV) demonstrated that BRCA status was not associated with residual tumor volume on multivariate analysis, which led the authors to conclude that improved survival in BRCA carriers was not attributable to an increased rate of optimal tumor debulking .Taken together, these studies consistently support a significantly improved survival in BRCA2 mutation carriers in comparison to sporadic OC patients, while for BRCA1 carriers the advantage, if any, seems smaller. in vivo was found, suggesting that mtDNA status might serve as a marker to predict therapeutic efficacy to PARP inhibitors. Further studies are warranted to confirm this correlation.A recent study demonstrates that BRCA1 mutations in breast cancer drive oxidative stress and glycolytic transformation of the tumor. Loss of BRCA1 function leads to hydrogen peroxide generation in both epithelial breast cancer cells and neighboring stromal fibroblasts and promotes the onset of a reactive glycolytic stroma. Importantly, these metabolic changes can be reversed by antioxidants, which potently induce cancer cell death . FurtherThe detection of a mutation in a patient with breast or ovarian cancer allows for identification of asymptomatic mutation carriers in the family. Such predictive genetic testing should be performed in the framework of a comprehensive genetic counseling process, aimed at facilitating informed decisions about testing and risk-reducing options.Carriers of BRCA mutations are at increased risk of both breast and ovarian cancer; such risks are consistently estimated to be higher in BRCA1 than in BRCA2 mutation carriers; moreover, the risk is higher for breast than for OC. Estimates of cancer risk are variable across different studies; nevertheless, two large meta-analyses, resulting from a total of 31 published studies, estimated an OC risk to age 70 of 49% for BRCA1 and 18% for BRCA2  and of 3In Europe and the United States, the onset of familial OC is generally reported to be 5 to 10 years earlier than that of sporadic OC, but the early onset is probably limited to BRCA1 mutation carriers. Indeed, Risch and colleagues reported a mean age at OC onset of 51.2 years for BRCA1 mutation carriers and of 57.5 years for BRCA2 positive women . SimilarOptions to reduce the risk of OC or fallopian tube cancer in BRCA mutations carriers include surveillance, chemoprevention, and surgery . NeverthRecommended surveillance should be performed with pelvic examination, transvaginal ultrasound, and CA-125 levels every 6 months beginning at age 30 or 5–10 years earlier than the youngest relative diagnosed with OC , 102. LiIn conclusion surveillance for OC in BRCA positive patients with pelvic exam, transvaginal ultrasound, and serum levels of CA-125 is not invasive, but its impact on early detection and mortality is low and may be associated with emotional distress .Since surveillance for ovarian, peritoneal, and fallopian tube cancer has not been proven to be effective, prophylactic salpingooophorectomy is actually recommended to BRCA positive women by the age of 40 years and upon completion of child bearing .Salpingooophorectomy in BRCA patients should be performed considering specific issues and adequate surgical details. Specific preoperative counselling and informed consents should be obtained . This suSalpingooophorectomy requires complete removal of the ovary and the fallopian tube; the procedure should therefore be preceded by an accurate periadnexal adhesiolysis to obtain clear margins and simple handling of the structures. Peritoneum should be entered laterally to the ovarian vessels and visualization of ureter, ovarian artery, and veins obtained. Ovarian vessels should be coagulated and cut 1-2 cm away from the ovary to obtain clear margins and to verify the absence of vascular and lymphatic involvement. The peritoneum next to the ovary has to be cut with large margins to identify microscopic neoplastic foci. Finally, uteroovarian ligament and fallopian tubes should be coagulated and sectioned as close to the uterine wall as possible .Fallopian tube and ovary are removed intact through an endobag to avoid spillage in the abdominal cavity and sent for frozen section. For logistic or organization issues frozen section could be missed, but the necessity of second surgery has to be discussed with the patient during preoperative counselling .The removal of the uterus is not mandatory when preoperative imaging has excluded endometrial or myometrial disease; the risk of remnant of the intramural fallopian tube is negligible, but it should be discussed with the patient. Hysterectomy simplifies hormone replacement therapy and eliminates the possible increased risk of endometrial cancer and the possible increased risk of serous carcinoma of the uterus , 113.Because of the tubal origin of serous ovarian carcinoma , some auFinally, single port procedures are now available and this low aesthetic impact surgical access could be proposed to BRCA 1 and BRCA 2 patients .The pathologic examination of specimens from risk-reducing salpingooophorectomy requires special attention. To this aim, the BRCA mutation status should be always shared with the pathologist, since in patients with benign gynecologic disease only one slide from the fallopian tube and ovary is normally reviewed, while in the setting of a known genetic predisposition to OC, most pathologists will submit the entirety of the fallopian tubes and ovaries for microscopic examination. The SEE-FIM (sectioning and extensively examining the fimbriated ends) protocol is widely used. Basically, this involves serially sectioning the tube meticulously, stopping before the fimbriae. The fimbria is amputated and sectioned longitudinally, thereby maximizing exposure. Deeper sections need to be obtained if foci of atypia are to be identified histologically. Foci of in situ or invasive occult carcinoma may be very subtle and are often less than 1 mm in maximum diameter. Microscopic occult carcinomas have been identified in these specimens in about 2% to 11% of BRCA mutation carriers, generally involving the tubal fimbriae –119.Literature data demonstrates that OC arising in BRCA1/2 mutation carriers have peculiar molecular, pathological, and clinical features. Since a nonnegligible proportion of newly diagnosed OC patients are expected to carry such mutations, BRCA1/2 mutational analysis would help, in the future, to tailor OC management according to BRCA status. In addition, this would allow the subsequent identification of asymptomatic carriers who would benefit from targeted interventions for high-risk women.


Hearing loss is a common feature in human aging. It has been argued that dysfunctions in central processing are important contributing factors to hearing loss during older age. Aging also has well documented consequences for neural structure and function, but it is not clear how these effects interact with those that arise as a consequence of hearing loss. This paper reviews the effects of aging and adult-onset hearing loss in the structure and function of cortical auditory regions. The evidence reviewed suggests that aging and hearing loss result in atrophy of cortical auditory regions and stronger engagement of networks involved in the detection of salient events, adaptive control and re-allocation of attention. These cortical mechanisms are engaged during listening in effortful conditions in normal hearing individuals. Therefore, as a consequence of aging and hearing loss, all listening becomes effortful and cognitive load is constantly high, reducing the amount of available cognitive resources. This constant effortful listening and reduced cognitive spare capacity could be what accelerates cognitive decline in older adults with hearing loss. Normal aging in humans is often accompanied by hearing loss  and normal hearing (N = 75) older adults in a baseline scan and a follow-up scan (on average) 6.4 years later. They found no significant differences between the groups at the baseline measure, but after the follow-up scan those with hearing impairment had an accelerated volume decline in the whole brain, and particularly in the right temporal lobe. In short, the use of more specific, hypothesis-driven definitions of regions of interest, in combination with longitudinal approaches, could aid some light on the mixed effects found when measuring gray matter changes as a consequence of hearing loss.In trying to understand discrepancies in the observed effects of hearing loss and aging on brain morphometry, there are three salient factors: (1) Lack of specificity when defining regions of interest, as explained above ; (2) Measuring the effects of high frequency hearing thresholds vs. average pure tone thresholds. Hearing loss tends to be greater for higher-frequency sounds, and as discussed by Eckert et al. , using tAnother non-invasive brain imaging technique to study brain structure is diffusion MRI (dMRI). This technique measures microstructural parameters, including fractional anisotropy (FA) and mean diffusivity (MD), which reflect properties such as the degree of density and orientation dispersion of neuronal fiber bundles Jones, . It alsoN = 12–15 per experimental group; Profant et al., Microstructural changes associated with hearing loss have been found in subcortical components of the auditory pathway [reduction in fractional anisotropy (FA) and increase in radial diffusivity], such as the lateral lemniscus and the inferior colliculus . By detecting changes in blood flow that occur as a consequence of neural activity, this technique allows indirect measurement of brain function in a non-invasive manner and with a spatial resolution of millimeters , and it is not clear whether it is mediated by age itself or by age-related hearing loss  , aids in adaptive control during word recognition  . In adults, the most prominent peaks are N1 (~100 ms post-stimulus onset) and P2 (~175–200 ms). Smaller peaks P1 and N2, preceding N1 and succeeding P2 respectively, are also often described , resulting in elevated thresholds and reduced amplitudes , Action on Hearing Loss (Project 598), and the Economic and Social Research Council of Great Britain (Grant RES-620-28-0002).The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Autism susceptibility candidate 2 gene (AUTS2) have been associated with a broad range of psychiatric illnesses including autism spectrum disorders, intellectual disability and schizophrenia. We previously demonstrated that the cytoplasmic AUTS2 acts as an upstream factor for the Rho family small GTPase Rac1 and Cdc42 that regulate the cytoskeletal rearrangements in neural cells. Moreover, genetic ablation of the Auts2 gene in mice has resulted in defects in neuronal migration and neuritogenesis in the developing cerebral cortex caused by inactivation of Rac1-signaling pathway, suggesting that AUTS2 is required for neural development. In this study, we conducted a battery of behavioral analyses on Auts2 heterozygous mutant mice to examine the involvement of Auts2 in adult cognitive brain functions. Auts2-deficient mice displayed a decrease in exploratory behavior as well as lower anxiety-like behaviors in the absence of any motor dysfunction. Furthermore, the capability for novel object recognition and cued associative memory were impaired in Auts2 mutant mice. Social behavior and sensory motor gating functions were, however, normal in the mutant mice as assessed by the three-chamber test and prepulse inhibition test, respectively. Together, our findings indicate that AUTS2 is critical for the acquisition of neurocognitive function.Mutations in the  Acquisition of the higher cognitive functions in mammalian brain is primarily accomplished by a sequence of neurodevelopmental events comprised of cell differentiation and migration followed by cell morphogenesis. The cellular changes include extension and branching of neurites and formation of synapses to assemble the proper neural circuits during brain development. Genetic defects involving these developmental processes lead to a variety of neuropsychiatric illness such as autism spectrum disorders (ASDs), intellectual disability (ID), schizophrenia and epilepsy.Autism susceptibility candidate 2 gene (AUTS2) by de novo balanced translocation, was originally associated with ASDs, but more studies have shown that it is also associated with a wide range of neurological diseases such as ID, attention deficit hyperactivity disorder (ADHD), dyslexia and epilepsy ; there were no effects of genotype , or interactions with genotype   relative to the opposite chamber containing the previous, now familiar mouse (stranger 1) , and there were no effects of genotype , or interactions with genotype   revealed a significant effect of prepulse intensity . There were no effects of genotype , or interaction of genotype   as indicators of sensorimotor processing. The data obtained from one  = 0.49]  left. Ho p<0.01,  right.Together, these results suggest that AUTS2 plays an important role for high-order brain functions such as anxiety-related emotional behaviors as well as in certain types of memory formation.AUTS2 was implicated as a promising candidate for multiple neurocognitive defects as well as developmental delay and epilepsy [AUTS2 coding regions, many other cases were exclusively structural variants in which AUTS2 was disrupted by genomic rearrangements including balanced translocation, inversions or deletions of AUTS2 non-coding regions, implying the possibility that other genes could be involved in the occurrence of the disorders. Recently, however, Beunders et al. have identified a significant number of individuals with ASD and/or ID having microdeletions in which at least one exon of AUTS2 was included [Auts2 alone sufficiently induced several behavioral abnormalities including decrease of exploratory behaviors in a new circumstance as well as the decrease of anxiety against aversive situations for mice. Furthermore, Auts2-deficient mice displayed learning defects related to recognition and associative memory, suggesting that AUTS2 contributes to the acquisition of neurocognitive brain function. Interestingly, Auts2-deficient mice exhibited higher responses against nociceptive (AUTS2 mutations.Many studies have previously demonstrated that epilepsy . In thosincluded . Consisticeptive  right whAUTS2 occasionally present with brain malformations including microcephaly and corpus callosum hypoplasia [Auts2-deficient mice do not show significant gross morphological and histological abnormalities in brain structure, several developmental processes including the cortical neuronal migration and subsequent axon development were impaired in both homozygotic and heterozygotic Auts2 mutant mice d[Auts2 mutant mice could be caused by the developmental abnormalities in the microstructure within the brain, which may hamper the establishment of proper neural circuits in the mature brain. Additionally, AUTS2 may also be involved in the postnatal neurodevelopmental processes such as synaptogenesis, synaptic neurotransmission and neural circuit formation because AUTS2 is continuously expressed in the mature mice brain. Future studies of Auts2 mutant mice will allow us to better understand the function of AUTS2 in these processes.Previous studies have demonstrated that mutations in the genes involved in the regulation of cytoskeleton organization were associated with mental retardation, epilepsy and learning disorders , 26. Forpoplasia , 35. In nt mice d. The def