Expression Of Key Genes In P38MAPK Signaling Pathway In Pre-Hierarchical Follicles Of Geese

Goose aquaculture industry is a rising industry in recent years,and the characteristics of seasonal goose production of geese are an important factor restricting the development of geese industry.The exploration of the follicle development mechanism of goose contributes to the continuous progress of the goose breeding industry in China.Present studies have shown that p38 MAPK signaling pathway has a certain regulatory effect on the follicular development of poultry.In this study,we selected the geese at the peak of egg production,collected pre-hierarchical follicles,and used immunohistochemical(IHC)technology to determine the expression sites of key genes in the pre-grade follicles on the pathway;using real-time quantitative PCR(qPCR)detections Expression of the target gene at the mRNA level;Western Blot technique was utilized to detect the expression of the target gene at the protein level,and the biological function of the target gene was analyzed preliminarily by significance test and correlation analysis.The research tests results are as follows:(1)The level of mRNA expression of key genes in the p38 MAPK signaling pathway in the pre-grade follicles of gooses was investigated by qPCR,and a significant test was performed.It was found that all the genes of interests in the study were expressed in five grades of pre-follicles,and there was a significant difference between the maximum and minimum levels of expression(p<0.05).Among them,the expression patterns of TGF-βI,TGF-βRII,Fas,and Fasl were increased first and then decreased,with the highest expression in MWF.The expression patterns of the target genes DAXX,ASK1,MKK3,MKK6,and p38α showed a trend of decreased first and then increased and then remained stable.The expression level in MWF was the lowest,among which ASK1 had the highest expression level in SYF,and the remaining genes were in pe follicles.The highest of the remaining genes was found in pe follicles.(2)The correlation analysis of qPCR test results showed that there was spatio-temporal difference in the correlations of genes on the p38 MAPK signaling pathway.In pe follicles,SWF,MWF,TGF-β and its receptors mainly regulated the signaling pathway,and inhibited the expression of genes involved in follicle development in the signaling pathway.In LWF and SYF,Fas and Fasl regulated the signaling pathway and promoted follicular development.(3)Localized analysis of the expressions of the target protein ASK1 and p38α in goose-grade pre-follicles by IHC experiments revealed that the two target proteins were present in the pre-follicular membrane cells(TC)and granular layer cells in 5 grades of goose.Positive expression sites were detected in(GC)and there was a certain spatial and temporal difference.(4)The protein levels of ASK1,MKK3,and p38α in the pre-grade follicles of Goose were analyzed by Western Blot.The results showed that the overall trend of the expression levels of the three target proteins in the pre-grade follicles was consistent with that in qPCR test results,but there were some differences,and the correlation analysis results were also different from the qPCR results.(5)Combined with the follicular development and gene expression patterns in the p38 MAPK signaling pathway,we found that p38 MAPK signaling pathway plays a certain role in promoting the pre-grade follicles of geese.TGF-βI,TGF-βRII,Fas,and Fasl contribute to follicular development.Inhibitory effects,DAXX,ASK1,MKK3,MKK6 and p38α promote follicular development.