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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。
Objective: To observe the expression of p75 neurotrophin receptor (P75NTR) in mouse testis and to explore its relationship with nestin. Methods: The expression of P75NTR and Nestin in Nestin-GFP transgenic mice testis were observed by immunofluorescence technique. The expression of P75NTR and Nestin in the testis were detected by immunofluorescence technique. Real-time PCR and flow cytometry were used to detect the expression of P75NTR at different ages Rat testis; expression of P75NTR-positive cells in culture medium of neural stem cells were cultured, through the directional differentiation of the ability to observe the neural differentiation. Results: Immunofluorescence staining showed that P75NTR and Nestin were localized in testicular stromal cells. Real-time PCR and flow cytometry results showed that P75NTR peaked in the testicular tissue of mice on the 14th day of life. The positive percentage of P75NTR in mouse testes at the 5th, 14th and 30th day after birth were (2.88 ± 0.52)%, (9.54 ± 1.81)% and (2.63 ± 0.43)% respectively. The sorted P75NTR positive cells were also co-localized Nestin and P75NTR, and have neuronal differentiation ability. CONCLUSION: P75NTR and Nestin are co-localized in testicular stromal cells in testes. These P75NTR positive cells have the ability of neural differentiation, suggesting that they originate from nerve ridges.