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与顶体生物发生和精子细胞伸长相关的新型睾丸表达的丝切蛋白IV

Novel testis-expressed profilin IV associated with acrosome biogenesis and spermatid elongation.

作者信息

Obermann Heike, Raabe Inka, Balvers Marga, Brunswig Bärbel, Schulze Wolfgang, Kirchhoff Christiane

机构信息

Department of Andrology, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany.

出版信息

Mol Hum Reprod. 2005 Jan;11(1):53-64. doi: 10.1093/molehr/gah132. Epub 2004 Dec 10.

Abstract

A novel profilin, named profilin IV, was cloned and characterized as a testicular isoform, distinct from the previously described testis-specific profilin III. Profilin IV showed only 30% amino acid identity with the other mammalian profilins; nevertheless, database searches produced significant alignments with the conserved profilin domain. Northern blot analysis and in situ transcript hybridization suggested that profilin IV, like profilin III, is transcribed in the germ cells. However, the timing of their expression during post-natal development of rat testis and in the rat spermatogenetic cycle was distinct. In the human testis, profilin IV mRNA expression correlates with the presence of germ cells suggesting that it may be a suitable molecular diagnostic parameter to supplement conventional histopathological diagnostics in the assessment of testicular biopsies. The predicted profilin IV protein was verified employing an anti-oligopeptide antibody. Western blot analysis detected an immunorelated testicular protein of approximately 14 kDa. Immunohistochemistry revealed an intracellular protein of the rat, the mouse and the human testis accumulating asymmetrically in the cytoplasm of round and elongating spermatids with its perinuclear location coinciding with the position of the developing acrosome-acroplaxome and the manchette. Profilin IV thus may regulate testicular actin cytoskeleton dynamics and play a role in acrosome generation and spermatid nuclear shaping.

摘要

一种名为原肌球蛋白IV的新型原肌球蛋白被克隆出来,并被鉴定为睾丸异构体,与先前描述的睾丸特异性原肌球蛋白III不同。原肌球蛋白IV与其他哺乳动物原肌球蛋白的氨基酸同一性仅为30%;然而,数据库搜索显示其与保守的原肌球蛋白结构域有显著比对。Northern印迹分析和原位转录杂交表明,原肌球蛋白IV与原肌球蛋白III一样,在生殖细胞中转录。然而,它们在大鼠睾丸出生后发育过程中和大鼠精子发生周期中的表达时间不同。在人类睾丸中,原肌球蛋白IV mRNA表达与生殖细胞的存在相关,这表明它可能是一种合适的分子诊断参数,可在评估睾丸活检时补充传统的组织病理学诊断。使用抗寡肽抗体验证了预测的原肌球蛋白IV蛋白。Western印迹分析检测到一种约14 kDa的与免疫相关的睾丸蛋白。免疫组织化学显示,大鼠、小鼠和人类睾丸中的一种细胞内蛋白不对称地积聚在圆形和伸长精子细胞的细胞质中,其核周位置与发育中的顶体-顶体板和纤丝鞘的位置一致。因此,原肌球蛋白IV可能调节睾丸肌动蛋白细胞骨架动力学,并在顶体生成和精子细胞核塑形中发挥作用。

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