Nestin-GFP转基因小鼠睾丸中P75NTR表达的研究

目的:观察小鼠睾丸中的p75神经营养因子受体(P75NTR)的表达规律,并探讨其与巢蛋白(nestin)之间的关系。方法:通过免疫荧光技术观察出生后第5天Nestin-GFP转基因小鼠睾丸组织中的P75NTR和巢蛋白的表达位置,并采用实时荧光定量PCR和流式细胞分选方法检测P75NTR在不同周龄小鼠睾丸组织中的表达量;对分选得到的P75NTR阳性细胞在神经干细胞的培养液中进行培养,通过定向分化技术观察其神经分化能力。结果:免疫荧光染色结果显示P75NTR和巢蛋白定位表达于睾丸间质细胞。实时荧光定量PCR和流式细胞分选结果显示,P75NTR在出生第14天小鼠睾丸组织中出现表达高峰。出生后第5、14、30天小鼠睾丸中P75NTR阳性百分率分别为(2.88±0.52)%、(9.54±1.81)%、(2.63±0.43)%,分选得到的P75NTR阳性细胞也共定位表达巢蛋白和P75NTR,并且具有神经分化能力。结论:睾丸组织中P75NTR和巢蛋白共定位表达于睾丸间质细胞,该类P75NTR阳性细胞具有神经分化能力,推测其起源于神经脊。

Expression of P75NTR in the testis of nestin-GFP transgenic mice

Objective: To explore the P75NTR expression in the mouse testis and its relationship with nestin.Methods: We observed the location of the expressions of P75NTR and nestin in the testis of the nestin-GFP transgenic mouse on postnatal day(PND) 5,14 and 30 using immunofluorescence,and detected the expression levels of P75NTR in the testicular tissue of mice in different age groups by real-time quantitative PCR(RTqPCR) and flow cytometry.Then we cultured the P75NTR positive cells in neural stem cell culture medium and observed their neuronal differentiation capacity by orientation differentiation.Results: Immunofluorescence showed the expressions of P75NTR and nestin in the Leydig cells of the mouse testis.RTqPCR and flow cytometry exhibited the peak of the P75NTR expression on PND 14.The positive rates of P75NTR were(2.88±0.52),(9.54±1.81) and(2.63±0.43)% on PND 5,14 and 30,respectively.The P75NTR positive cells obtained also expressed nestin and P75NTR and had the capacity of neuronal differentiation.Conclusion: P75NTR and nestin are co-expressed in the Leydig cells of the mouse testis,and the P75NTR positive cells have the ability of neural differentiation,which is presumably attributed to neural crest cells.