Expression of transcriptional factor EB (TFEB) in differentiating spermatogonia potentially promotes cell migration in mouse seminiferous epithelium.
Expression of transcriptional factor EB (TFEB) in differentiating spermatogonia potentially promotes cell migration in mouse seminiferous epithelium.
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转录因子 EB (TFEB) 在分化精原细胞中的表达可能促进小鼠生精上皮细胞的迁移
DOI：
10.1186/s12958-018-0427-x
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发表时间：
2018-10-25
期刊：
影响因子：
--
通讯作者：
Xu C
中科院分区：
文献类型：
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作者：
Liu Y;Hu Y;Wang L;Xu C
研究背景精子发生是一个复杂的过程，包括精原细胞在曲细精管中自我更新和分化为成熟精子细胞。在精子发生过程中，生殖细胞从基底膜迁移，穿过血睾屏障（BTB），最终到达生精上皮的腔侧。然而，调节生殖细胞迁移的机制仍不清楚。在这项研究中，我们专注于转录因子EB（TFEB）的表达和功能，一个主调节溶酶体生物合成，自噬和内吞作用，在spermatogenes.MethodsThe TFEB在小鼠睾丸的表达模式进行了研究，通过Western印迹和免疫组化分析。使用基于特定细胞表面标记的磁激活细胞分选从睾丸中分离出未分化的精原细胞或分化的精原细胞。用100 nM维甲酸（RA）诱导精原细胞分化。使用shRNA敲低细胞中的TFEB。通过免疫荧光、qRT-PCR和Western印迹检测TFEB表达。通过transwell迁移试验和伤口愈合试验测定细胞迁移，所述细胞迁移应用于永生化精原细胞GC-1细胞系。结果在睾丸发育期间，TFEB在睾丸中的表达在产后7天（dpp）至14 dpp期间迅速增加，而在成年睾丸中，在生精上皮周期的VI至VIII阶段，它主要定位于精原细胞核中。因此，TFEB被观察到主要在分化的精原细胞中表达，并被RA处理激活进行核转位。此外，敲低TFEB表达的RNAi并不影响精原细胞分化，但显着减少细胞迁移GC-1 cells.ConclusionThese研究结果意味着，区域不同的表达和激活的TFEB是强烈相关的RA信号，因此可能会促进细胞迁移跨越BTB和运输沿着生精上皮。
BackgroundSpermatogenesis is a complex process involving the self-renewal and differentiation of spermatogonia into mature spermatids in the seminiferous tubules. During spermatogenesis, germ cells migrate from the basement membrane to cross the blood-testis barrier (BTB) and finally reach the luminal side of the seminiferous epithelium. However, the mechanism for regulating the migration of germ cells remains unclear. In this study, we focused on the expression and function of transcriptional factor EB (TFEB), a master regulator of lysosomal biogenesis, autophagy and endocytosis, in spermatogenesis.MethodsThe expression pattern of the TFEB in mouse testes were investigated by Western blotting and immunohistochemistry analyses. Either undifferentiated spermatogonia or differentiating spermatogonia were isolated from testes using magnetic-activated cell sorting based on specific cell surface markers. Differentiation of spermatogonia was induced with 100 nM retinoic acid (RA). shRNA was used to knock down TFEB in cells. TFEB expression was detected by immunofluorescence, qRT-PCR, and Western blotting. Cell migration was determined by both transwell migration assay and wound healing assay applied to a cell line of immortalized spermatogonia, GC-1 cells.ResultsDuring testicular development, TFEB expression was rapidly increased in the testes at the period of 7 days post-partum (dpp) to 14 dpp, whereas in adult testis, it was predominantly localized in the nucleus of spermatogonia at stages VI to VIII of the seminiferous epithelial cycle. Accordingly, TFEB was observed to be mainly expressed in differentiating spermatogonia and was activated for nuclear translocation by RA treatment. Moreover, knockdown of TFEB expression by RNAi did not affect spermatogonial differentiation, but significantly reduced cell migration in GC-1 cells.ConclusionThese findings imply that regionally distinct expression and activation of TFEB was strongly associated with RA signaling, and therefore may promote cell migration across the BTB and transport along the seminiferous epithelium.