The Expressions Of NSE、SYP、S100B And 5-HT In Htert-caprine Luteal Cells

Our previous studies revealed that there were a certain amount of diffuse neuroendocrine cells in goat’s and bovine’s CL,meanwhile,the DNE cells’specific markers—neuron specific enolase(NSE),synaptophysin(SYP),5-hydroxytryptamine(5-HT),and S-100 protein existed with CL’s formation and disappeared with CL’s regression,which strongly suggested that the four markers may involve in the CL cellular formation,evolution and regression.In order to explore each role of these four markeres in the goat CL cells,the current work is going to detect the expressions of these four markers in hTERT-CLCs,then compare each expression result with that in the purified goat luteal cells using fluorescence immunocytochemistry staining and real-time fluorescent quantitative PCR.We hope this hTERT-CLCs could be a good cell model in investigating the functions of these four markers in vitro.The experimental results are as follows:1.This study has demonstrated that Percoll density gradient centrifugation was superior to the method of differential detachment in cell purification efficiency and in maintenance of cell proliferation activity.We are confident that the Percoll discontinuous density gradient centrifugation method is a simple and economical method to purify the goat luteal cells and this method can be widely used in the future.2.The dairy goats SYP and S100B gene CDS were firstly identified in the present study.The GenBank sequence ID was KC244281.1 and KC244280.1,respectively.3.NSE and SYP were expressed in hTERT-CLCs after cultured 48 h,and the expression of NSE in hTERT-CLCs was lower than that in normal goat luteal cells(P<0.01).5-HT was only expressed in parts of hTERT-CLCs’cytoplasm.4.NSE,SYP,S100B and 5-HT were expressed in normal goat luteal cells after cultured 48 h,and the order of mRNA expression quantity was:NSE>S100B>SYP.To sum up:NSE and 5-HT were expressed in hTERT-CLCs,but no expression of SYP and S100B were detected.The hTERT-CLCs will be a good cell model in investigating the functions of NSE and 5-HT in vitro.