Study Of Significance And Expressions Of PEA3 And GDNF In Colon Auerbach Plexus Of Mouse

Abstract Enteric nervous system(ENS) was composed by neurons located in gastrointestinal tract (include pancreas and gallbladder),transmitteres and proteins as well as sustentacular cells,These neurons traceabled nervous cres(tNC). Nervous system located in gastrointestinal tract,was composed by two groups neurons as well as fab web,one is situated between longitudinal muscle and circleround muscle,named auerbach plexus;the orther is situated between mucous layer and circleround muscle,named meissner plexus. the former main related on enteric active,the latter main related on secreting and absorptive function. When auerbach plexus was developmental disorder,intestinal tract motor function was cacergasia,then lead to constipation and so on. For instance Hirchsprung's disease(HD).Polymoma virus enhancer-3(PEA3),belong to ETS transcription factor gene family. It coding and born protein and to bring into transference and differentiation of NC , especially to motoneuron. It was said that glial cell line-derived neurotrophic factor(GDNF)act as the upstream gene of PEA3 to influence The expressions of it. And found that neurocyte migration disorder both in PEA3 and GDNF mutants. Departed study discovered,GDNF express defecting sent to nerves precursor celles prematurity braid dead leading to ENS abnormity,It was one of important factors of HD. SOX-10 gene sensitived to HD,Gene knock-out mouse showed many clinical symptoms of HD. But if PEA3 and GDNF produce a marked effect in HD and HAD , there are no reports now. We used immunohistochemical method to investigate the expressions of PEA3,GDNF in myenteric nerve plexus,to investigate the expressions of PEA3 in myenteric nerve plexus of SOX-10 gene knock out mice,then to investigate etiopathogenisis of HD and HAD further.This experiment was divided into two parts to study nosogenesis of HD and HAD: Primary part,we used thirteen majority female Kunming mice,BM 50～60g,coitioning with males,BM 80g, confirming potentia generandi one to one,every early morning,we examed vagina,if found female suppos or great spermatozoas,we considered mating success and settled gravidity the first day. To obtain colon of mouse preimplantatigene additionembryos of Different gestational age 10 days,14 days,19 days. Paraffin imbedding. Per-lump sect serial sections 5 to remain 1,thick 6 micrometer, Immunohistochemistry:using Rat PEA3 monoclonal antibody(SANTA CRUZ,dilution 1:400),Rat GDNF monoclonal antibody(BOST,dilution 1:100) as first antibody,SABC. DAB dyeing. Using PBS to take the place of serum to contrast,As negative result. To observe the distribution of PEA3 and GDNF in colon auerbach plexus development process,to investigate the relation of PEA3 and HD dyskinesis;The second part,To obtain ascending colon and igmoid colon of five knock out mice and one normal mice, paraffin imbedding,Per-lump sect serial sections 3,thick 6 micrometer,named NO.1,NO.2,NO.3,NO.1 dyeing,NO.2 Immunohistochemistry dyeing,NO.3 TB dyeing. The last is the same with the Primary part. To investigate the Expressions difference of PEA3 protein in colon between knock out mice and normal mice;To investigate the etiopathogenisis of HD further. Experimental result:Primary part,We found that optical density mean value of PEA3 and GDNF were low in the colon development nonage, GDNF achieved to peak value in the colon development metaphase,then descended; and the expressions of PEA3 was increasing when the fetal age was raising,and the expressions of PEA3 and GDNF proteins in different fetal age kept coherence. Through statistical analysis, x±s, n=5,10-days group compared with 14-days group: P<0.01; 14-days group compared with 19-days group: P<0.01; 10-days group compared with 19-days group: P<0.01. The second part,The diversity in quantitatively of PEA3 masccline cell in sascending colon between these two group is not obvious,in colon sigmoideum was decreased, but the quantitaty of PEA3 positive particle in knock out mice is lesser than that in normal mice. Throungh photodensitometry and statistics analysis,we can find that the expressions difference of PEA3 protein in colon between knock out mice and normal mice have statistics sense(P<0.01) .These result hinted that,The expressions of Pea3 gene in colon of mouse preimplantations embryos kept equal pace with GDNF in colon development nonage and metaphase,And possiblly,GDNF act as the upstream gene of PEA3 to influence the expressions of it, all of these two genes participate the morbidity of enteric nervous development. And possiblely , PEA3 is one of the etiopathogenisises of HD and HAD. The PEA3 genes play an important role in the neural development of colon,SOX-10 gene impact the Expressions of PEA3.