XXXX as a Model to Identify Translation Impairment.

@entity71576 @entity1 @entity1238 @entity16 @entity563 @entity1889
Protein synthesis is a fundamental process to gene expression impacting diverse biological processes notably adaptation to environmental conditions. The initiation step, which involves the assembly of the ribosomal subunits at the mRNA initiation codon, involved initiation factor including @entity71576 . Defects in this rate limiting step of translation are linked to diverse disorders. To study the potential consequences of such deregulations, @entity1238 oocytes constitute an attractive model with high degrees of conservation of essential cellular and molecular mechanisms with @entity1 . In addition, during meiotic maturation, oocytes are transcriptionally repressed and all necessary proteins are translated from preexisting, maternally derived mRNAs. This inexpensive model enables exogenous mRNA to become perfectly integrated with an effective translation. Here is described a protocol for assessing translation with a factor of interest (here @entity71576 ) using stored maternal mRNA that are the first to be polyadenylated and translated during oocyte maturation as a physiological readout. At first, mRNA synthetized by in vitro transcription of plasmids of interest (here @entity71576 ) are injected in oocytes and kinetics of oocyte maturation by Germinal Vesicle Breakdown detection is determined. The studied maternal mRNA target is the @entity563 / @entity1889 -protein-kinase mos. Its polyadenylation and its subsequent translation are investigated together with the expression and phosphorylation of proteins of the mos signaling cascade involved in oocyte maturation. Variations of the current protocol to put forward translational defects are also proposed to emphasize its general applicability. In light of emerging evidence that aberrant protein synthesis may be involved in the pathogenesis of @entity16 , such a model provides the opportunity to easily assess this impairment and identify new targets.
