Planar Cell Polarity (PCP) Protein Vangl2 Regulates Ectoplasmic Specialization Dynamics via Its Effects on Actin Microfilaments in the Testes of Male Rats
Planar Cell Polarity (PCP) Protein Vangl2 Regulates Ectoplasmic Specialization Dynamics via Its Effects on Actin Microfilaments in the Testes of Male Rats
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DOI：
10.1210/en.2015-1987
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发表时间：
2016-05-01
期刊：
影响因子：
4.8
通讯作者：
Cheng, C. Yan
中科院分区：
文献类型：
--
作者：
Chen, Haiqi;Mruk, Dolores D.;Cheng, C. Yan
平面细胞极性（PCP）蛋白赋予上皮平面内的细胞场（例如，伸长/伸长的精子细胞）的极化，所述上皮平面例如精子发生期间的小管的生精上皮。在成年大鼠睾丸中，发现支持细胞和生殖细胞表达PCP核心蛋白（例如，货车Gogh样2 [Vangl 2]）、效应子、配体和信号蛋白。Vangl 2主要由Sertoli细胞表达，定位于睾丸特异性的、富含肌动蛋白的外质特化（ES），位于近腔室中的Sertoli-精子细胞界面处，也位于血-睾丸屏障（BTB）处的Sertoli-Sertoli界面处，并在结构上与肌动蛋白、Ncadherin和另一种PCP/极性蛋白Scribble相互作用。Vangl 2敲低（KD）在体外培养的支持细胞与建立紧密连接通透性屏障导致BTB收紧，而其过表达使用全长cDNA构建扰乱屏障功能。这些变化是通过改变组织肌动蛋白微丝在支持细胞的ES，涉及肌动蛋白调节蛋白，表皮生长因子受体途径底物8，肌动蛋白相关蛋白3，和Scribble，这反过来又影响了粘附蛋白复合物的功能，在精子发生的上皮周期的ES。使用Polyplus in vivo-jetPEI试剂作为转染介质，通过高效的RNA干扰在体内沉默睾丸中的Vangl 2，Vangl 2 KD导致上皮中ES处的F-肌动蛋白组织的变化，阻碍精子细胞和吞噬体转运以及精子细胞极性、减数分裂和BTB动力学。例如，第19级精子细胞与第9级和第10级精子细胞一起在IX-X期小管中保持嵌入上皮中。总之，PCP蛋白Vangl 2是通过其对睾丸中肌动蛋白微丝的作用的ES调节剂。
Planar cell polarity (PCP) proteins confer polarization of a field of cells (eg, elongating/elongated spermatids) within the plane of an epithelium such as the seminiferous epithelium of the tubule during spermatogenesis. In adult rat testes, Sertoli and germ cells were found to express PCP core proteins (eg, Van Gogh-like 2 [Vangl2]), effectors, ligands, and signaling proteins. Vangl2 expressed predominantly by Sertoli cells was localized at the testis-specific, actin-rich ectoplasmic specialization (ES) at the Sertoli-spermatid interface in the adluminal compartment and also Sertoli-Sertoli interface at the blood-testis barrier (BTB) and structurally interacted with actin, Ncadherin, and another PCP/polarity protein Scribble. Vangl2 knockdown (KD) by RNA interference in Sertoli cells cultured in vitro with an established tight junction-permeability barrier led to BTB tightening, whereas its overexpression using a full-length cDNA construct perturbed the barrier function. These changes were mediated through an alteration on the organization actin microfilaments at the ES in Sertoli cells, involving actin-regulatory proteins, epidermal growth factor receptor pathway substrate 8, actin-related protein 3, and Scribble, which in turn affected the function of adhesion protein complexes at the ES during the epithelial cycle of spermatogenesis. Using Polyplus in vivo-jetPEI reagent as a transfection medium to silence Vangl2 in the testis in vivo by RNA interference with high efficacy, Vangl2 KD led to changes in F-actin organization at the ES in the epithelium, impeding spermatid and phagosome transport and spermatid polarity, meiosis, and BTB dynamics. For instance, step 19 spermatids remained embedded in the epithelium alongside with step 9 and 10 spermatids in stages IX-X tubules. In summary, the PCP protein Vangl2 is an ES regulator through its effects on actin microfilaments in the testis.