Abnormal serum vitamin A levels and retinoic acid receptor α expression patterns in children with anorectal malformation.
BACKGROUND
Anorectal malformation (ARM) is known to be associated with maldevelopment of the enteric nervous system (ENS), and vitamin A (VA) and its metabolite retinoic acid (RA) play important roles in ENS development. Thus, our aim was to investigate serum VA levels in ARM newborns and RA receptor (RAR) expression in the rectum of ARM patients and animal models.


METHODS
Serum VA concentrations were detected in newly diagnosed ARM neonates (n = 32) and neonates with non-alimentary tract malformations (n = 30). Intestinal specimens were divided into three groups: rectum from ARM patients (n = 30), colon from a stoma (n = 30) and rectum from controls (n = 4). RAR mRNA expression was evaluated by RT-qPCR. Rectum specimens from ARM patients were divided into two groups by postoperative pathology: the normal and lesion ganglion cell groups. Immunohistochemistry and Western blot were employed to detect RARα protein expression in rectum specimens. In addition, the ARM mouse model was induced by all-trans retinoid acid (ATRA), and the expression levels of RARα and the neuronal marker NeuN in the rectum of mice on embryonic day 16.5-18.5 (E16.5-18.5) were investigated.


RESULTS
The serum concentration of VA in ARM neonates was lower than that in control neonates (P < 0.0001), and RARα mRNA expression was lower in the rectum specimens from ARM patients than in the colon specimens from a stoma and the rectum specimens from controls (P < 0.05); there was no significant difference between the colon from a stoma and the rectum from controls. RARα protein was expressed in the nucleus of ganglion cells and nerve fibers, and RARα protein expression in the lesion ganglion cell group was significantly lower than that in the normal ganglion cell group (P < 0.01). Compared with the control mice, ARM mice at E16.5-18.5 showed decreased fluorescence intensity of RARα and NeuN in the rectum. RARα and NeuN mRNA expression in the rectum on E16.5-18.5 was lower in ARM mice than in control mice (P < 0.05).


CONCLUSION
Serum VA concentration and the RARα expression pattern are abnormal in the rectum in ARM and may contribute to the ENS maldevelopment in ARM.
['Wang|Zhili|Z|', 'Wang|Quan|Q|', 'Gu|Chengchao|C|', 'Zhang|Jingjie|J|', 'Wang|Yi|Y|']
D000818:Animals D000831:Animals, Newborn D000071056:Anorectal Malformations D002675:Child, Preschool D004195:Disease Models, Animal D005260:Female D005786:Gene Expression Regulation D006801:Humans D007150:Immunohistochemistry D007223:Infant D008297:Male D051379:Mice D008810:Mice, Inbred C57BL D012313:RNA D000072482:Retinoic Acid Receptor alpha D014801:Vitamin A
Vitamin A Retinoic acid receptor Anorectal malformation Enteric nervous system