Yang Zhe, Liu Zhi, Yang Yanping, Dai Yanping, Gao Xiaoqin
Department of Histology and Embryology, School of Basic Medicine, Guizhou Medical University, Guiyang, Guizhou 550025, P.R. China.
Department of Urinary Surgery, The Second Affiliated Hospital of Guizhou Medical University, Guizhou Medical University, Kaili, Guizhou 556000, P.R. China.
Exp Ther Med. 2021 Aug;22(2):847. doi: 10.3892/etm.2021.10279. Epub 2021 Jun 7.
Sertoli cells (SCs) are an important component of spermatogenic tubules. The blood-testis barrier (BTB) is composed of SCs and is necessary for the development and maturity of spermatogenic cells. When the tight connection between SCs is destroyed, the BTB loses its integrity, leading to impaired spermatogenesis. Polypyrimidine tract-binding protein 1 (PTBP1) is a key protein involved in precursor mRNA splicing and selective splicing events, which directly affects tumor cell proliferation and influences the formation of the blood-tumor barrier by regulating the expression levels of tight junction-associated proteins. The present study revealed that the expression of PTBP1 was downregulated following a decrease in spermatogenic activity at the phase of senescence. TM4 cells were transfected with lentivirus-short hairpinRNA-PTBP1 to evaluate the effect of silencing PTBP1 on the expression levels of tight junction proteins and the integrity of tight junctions between adjacent SCs. Western blot analysis indicated that the expression levels of Zonula occludens 1, occludin and claudin-5 decreased significantly due to silencing of PTBP1 in SCs. Through detecting trans-epithelial electrical resistance, it was revealed that silencing of PTBP1 broke the integrity of tight junctions between adjacent SCs. The results suggested that PTBP1 maintained the integrity of the BTB by promoting the expression levels of tight junction-associated proteins and revealed the possible mechanism of PTBP1 in regulating spermatogenesis.
支持细胞(SCs)是生精小管的重要组成部分。血睾屏障(BTB)由支持细胞构成,对生精细胞的发育和成熟至关重要。当支持细胞之间的紧密连接被破坏时,血睾屏障失去完整性,导致精子发生受损。多聚嘧啶序列结合蛋白1(PTBP1)是一种参与前体mRNA剪接和选择性剪接事件的关键蛋白,它直接影响肿瘤细胞增殖,并通过调节紧密连接相关蛋白的表达水平影响血瘤屏障的形成。本研究表明,在衰老阶段生精活性降低后,PTBP1的表达下调。用慢病毒短发夹RNA-PTBP1转染TM4细胞,以评估沉默PTBP1对紧密连接蛋白表达水平及相邻支持细胞间紧密连接完整性的影响。蛋白质免疫印迹分析表明,由于支持细胞中PTBP1的沉默,紧密连接蛋白1、闭合蛋白和Claudin-5的表达水平显著降低。通过检测跨上皮电阻发现,PTBP1的沉默破坏了相邻支持细胞间紧密连接的完整性。结果表明,PTBP1通过促进紧密连接相关蛋白的表达水平维持血睾屏障的完整性,并揭示了PTBP1调节精子发生的可能机制。