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小鼠生精上皮中 VE-cadherin 介导的精子细胞-支持细胞连接的动态变化。

Dynamic of VE-cadherin-mediated spermatid-Sertoli cell contacts in the mouse seminiferous epithelium.

机构信息

Department of Biosciences, University of Milan, Via Celoria 26, 20133, Milan, Italy.

Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy.

出版信息

Histochem Cell Biol. 2018 Aug;150(2):173-185. doi: 10.1007/s00418-018-1682-9. Epub 2018 May 25.

Abstract

Spermatids are haploid differentiating cells that, in the meantime they differentiate, translocate along the seminiferous epithelium towards the tubule lumen to be just released as spermatozoa. The success of such a migration depends on dynamic of spermatid-Sertoli cell contacts, the molecular nature of which has not been well defined yet. It was demonstrated that the vascular endothelial cadherin (VEC) is expressed transitorily in the mouse seminiferous epithelium. Here, we evaluated the pattern of VEC expression by immunohistochemistry first in seminiferous tubules at different stages of the epithelial cycle when only unique types of germ cell associations are present. Changes in the pattern of VEC localization according to the step of spermatid differentiation were analysed in detail using testis fragments and spontaneously released germ cells. Utilizing the first wave of spermatogenesis as an in vivo model to have at disposal spermatids at progressive steps of differentiation, we checked for level of looser VEC association with the membrane by performing protein solubilisation under mild detergent conditions and assays through VEC-immunoblotting. Being changes in VEC solubilisation paralleled in changes in phosphotyrosine (pY) content, we evaluated if spermatid VEC undergoes Y658 phosphorylation and if this correlates with VEC solubilisation and spermatid progression in differentiation. Altogether, our study shows a temporally restricted pattern of VEC expression that culminates with the presence of round spermatids to progressively decrease starting from spermatid elongation. Conversely, pY658-VEC signs elongating spermatids; its intracellular polarized compartmentalization suggests a possible involvement of pY658-VEC in the acquisition of spermatid cell polarity.

摘要

精子细胞是处于分化状态的单倍体细胞,在分化过程中,它们沿着生精上皮向管腔方向移动,最终被释放为精子。这种迁移的成功取决于精子细胞与支持细胞之间的动态接触,但其分子本质尚未得到很好的定义。已经证明,血管内皮钙黏蛋白(VEC)在小鼠生精上皮中短暂表达。在这里,我们首先通过免疫组织化学评估了 VEC 在生精小管中的表达模式,这些小管处于上皮周期的不同阶段,此时只存在独特类型的生殖细胞联系。通过使用睾丸片段和自发释放的生殖细胞,详细分析了 VEC 定位模式根据精子细胞分化阶段的变化。利用第一波精子发生作为体内模型,使我们能够在分化的渐进步骤中获得精子细胞,我们通过在温和的去污剂条件下进行蛋白质溶解并通过 VEC-免疫印迹进行检测,检查 VEC 与膜的松散结合程度是否发生变化。由于 VEC 溶解的变化与磷酸酪氨酸(pY)含量的变化平行,我们评估了精子细胞 VEC 是否经历 Y658 磷酸化,以及这是否与 VEC 溶解和精子细胞分化中的进展相关。总之,我们的研究显示出 VEC 表达的时间限制模式,该模式在圆形精子细胞的存在时达到顶峰,然后从精子细胞伸长开始逐渐减少。相反,pY658-VEC 标记伸长的精子细胞;其细胞内极化的分区化表明 pY658-VEC 可能参与了精子细胞极性的获得。

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