METHODS: The testes of d6, d8, d17 and adult mice were collected, single cell suspension was prepared by combinatorial enzyme digestion, spermatogenic cells were isolated by BSA density gradient method, and Zfx expression was detected by real-time quantitative polymerase chain reaction (qRT-PCR) and Western Blot (WB).
RESULTS: Single cell suspension prepared by combination enzyme digestion method and density gradient method laid with BSA can obtain various types of spermatogenic cells with purity>85%; The expression level of the Zfx gene is low in primitive type A spermatogonia, type A spermatogonia, and type B spermatogonia, whereas it is high in preleptotene spermatocytes, pachytene spermatocytes, and round spermatid cells. It is not expressed in elongating spermatids and mature sperm.
CONCLUSIONS: Zfx gene exhibits periodic expression in various levels of spermatogenic cells and may be an important transcription factor involved in regulating meiosis in spermatogenic cells.
方法:收集d6、d8、d17和成年小鼠的睾丸,通过组合酶消化制备单细胞悬液,用BSA密度梯度法分离生精细胞,实时定量聚合酶链反应(qRT-PCR)和WesternBlot(WB)检测Zfx的表达。
结果:采用酶消化法和密度梯度法结合BSA制备的单细胞悬液可获得纯度>85%的各种类型的生精细胞;Zfx基因在原始A型精原细胞中的表达水平较低,A型精原细胞,和B型精原细胞,而它在前瘦素精母细胞中含量高,粗线质精母细胞,和圆形精子细胞。它在延伸的精子细胞和成熟的精子中不表达。
结论:Zfx基因在不同水平的生精细胞中呈周期性表达,可能是调控生精细胞减数分裂的重要转录因子。