Study On The Correlation Between Cadherin-22 And AKT3 In Mouse Female Germline Stem Cells

Germline stem cells are a kind of germ cells with capacities of self-renewal and differentiation,which can differentiated into functional gametes.It includes spermatogonial stem cells(SSCs)and female germline stem cells(FGSCs).The self-renewal and differentiation potential of germline stem cells is crucial to the individual’s reproductive ability.However,the existing research on the self-renewal and differentiation of female germline stem cells is limited,and the mechanism of self-renewal and differentiation is not clear.To further reveal the intrinsic mechanism of self-renewal and differentiation of female germline stem cells,this study began with the link of AKT3 and CDH22 in FGSCs,and explored the role of both factors in the self-renewal of female germline stem cells.First,we analyzed the differential gene expression profile DEG database of FGSCs and SSCs by bioinformatics,and selected some genes highly expressed in FGSCs,and further studied the proteins related to self-renewal of FGSCs.Screening analysis predicted that the AKT3 gene might play a role in self-renewal of FGSCs.In order to verify this hypothesis,we isolated and purified female germline stem cells for stable passage and proliferation in vitro.Next,we detected the high expression levels of CDH22 and AKT3 in the ovary at different stages of the mouse.using immunohistochemistry.We hypothesized that AKT3 has a similar effect on self-renewal of FGSCs with that in SSCs.The results of immunofluorescence BrdU/MVH double labeling assay showed that the proliferation of FGSCs was significantly inhibited after interfering with the expression of AKT3.This is consistent with our proposal.It is known that CDH22 can regulate the self-renewal of FGSCs through JAK-STAT signaling pathway.In order to verify whether AKT3 can interact with CDH22 and regulate the self-renewal of FGSCs,we analyzed the relationship between these two molecules by immunoprecipitation experiments.The results of co-immunoprecipitation showed that there was an interaction between CDH22 protein and PI3K.This result preliminarily proved that PI3K may be an intermediate molecule of CDH22 regulating AKT3.CDH22 protein can affect the activity of AKT3 through PI3K,thus affecting the proliferation of female germline stem cells.In summary,this study uses FGSCs as an experimental model,combines with bioinformatics analysis and prediction,to study the key factors affecting the mechanism of FGSCs self-renewal.However,the exact mechanism of interaction between CDH22 with AKT3 and how AKT3 plays a role in promoting self-renewal of FGSCs require more further research.This study primarily provides theoretical support for revealing the molecular mechanism of self-renewal of FGSCs,and also helpful for ovarian dysfunction caused by female animals in livestock production.