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全长和截短的Fyn酪氨酸激酶转录本在精子发生过程中的表达以及在顶体生物发生和受精过程中的定位。

Expression of full-length and truncated Fyn tyrosine kinase transcripts and encoded proteins during spermatogenesis and localization during acrosome biogenesis and fertilization.

作者信息

Kierszenbaum Abraham L, Rivkin Eugene, Talmor-Cohen Anat, Shalgi Ruth, Tres Laura L

机构信息

Department of Cell Biology and Anatomy, The Sophie Davis School of Biomedical Education/The City University of New York Medical School, New York, New York 10031, USA.

出版信息

Mol Reprod Dev. 2009 Sep;76(9):832-43. doi: 10.1002/mrd.21049.

Abstract

We report that full-length and truncated transcripts of Fyn tyrosine protein kinase are expressed during testicular development. Truncated Fyn (tr-Fyn) transcripts encode a 24 kDa protein with a N-terminal (NT) domain, a complete Src homology (SH) 3 domain and an incomplete SH2 domain. The kinase domain is missing in tr-Fyn. In contrast, full-length Fyn transcripts encode a 59-55 kDa protein. Fractionated spermatids by centrifugal elutriation express tr-Fyn transcripts and protein, but not full-length Fyn transcripts and protein. Neither full-length Fyn nor tr-Fyn transcripts and encoded proteins are detected in elutriated pachytene spermatocytes. Sertoli cells express full-length and truncated Fyn throughout testicular development. In contrast, sperm contain full-length Fyn transcripts and protein but not the truncated form. tr-Fyn protein is visualized at the cytosolic side of Golgi membranes, derived proacrosomal vesicles, along the outer acrosome membrane and the inner acrosome membrane-acroplaxome complex anchoring the acrosome to the spermatid nuclear envelope. Fyn and phosphotyrosine immunoreactivity coexist in the tail of capacitated sperm. During fertilization, the Fyn-containing acroplaxome seen in the egg-bound and egg-fused sperm is no longer detected upon decondensation of the sperm nucleus. tr-Fyn expands the catalog of truncated tyrosine protein kinases expressed during spermiogenesis. We suggest that the NT and SH3 domains of tr-Fyn may recruit adaptor and effector proteins, in particular GTPase activating proteins, required for acrosome-acroplaxome biogenesis, acroplaxome F-actin dynamics and Sertoli cell function. During fertilization, full-length Fyn in the acroplaxome may contribute to a transient local signaling burst during the early events of sperm-egg interaction.

摘要

我们报告称,Fyn酪氨酸蛋白激酶的全长和截短转录本在睾丸发育过程中表达。截短的Fyn(tr-Fyn)转录本编码一种24 kDa的蛋白质,该蛋白质具有一个N端(NT)结构域、一个完整的Src同源(SH)3结构域和一个不完整的SH2结构域。tr-Fyn中缺少激酶结构域。相比之下,全长Fyn转录本编码一种59 - 55 kDa的蛋白质。通过离心淘洗分离的精子细胞表达tr-Fyn转录本和蛋白质,但不表达全长Fyn转录本和蛋白质。在淘洗得到的粗线期精母细胞中未检测到全长Fyn和tr-Fyn转录本及编码蛋白。支持细胞在整个睾丸发育过程中都表达全长和截短的Fyn。相比之下,精子含有全长Fyn转录本和蛋白质,但不含有截短形式。tr-Fyn蛋白在高尔基体膜的胞质侧、衍生的前顶体小泡、顶体膜外侧以及将顶体锚定到精子细胞核膜的顶体膜内侧 - 顶体下板复合体中可见。Fyn和磷酸酪氨酸免疫反应性在获能精子的尾部共存。在受精过程中,当精子核解聚时,在与卵子结合和融合的精子中可见的含Fyn的顶体下板不再被检测到。tr-Fyn扩展了精子发生过程中表达的截短酪氨酸蛋白激酶的种类。我们认为,tr-Fyn的NT和SH3结构域可能募集衔接蛋白和效应蛋白,特别是顶体 - 顶体下板生物发生、顶体下板F - 肌动蛋白动力学和支持细胞功能所需的GTP酶激活蛋白。在受精过程中,顶体下板中的全长Fyn可能在精卵相互作用的早期事件中促成短暂的局部信号爆发。

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