Novel aspect of perinuclear theca assembly revealed by immunolocalization of non-nuclear somatic histories during bovine spermiogenesis
Novel aspect of perinuclear theca assembly revealed by immunolocalization of non-nuclear somatic histories during bovine spermiogenesis
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DOI：
10.1095/biolreprod.104.030445
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发表时间：
2004-10-01
影响因子：
3.6
通讯作者：
Oko, RJ
中科院分区：
文献类型：
--
作者：
Tovich, PR;Sutovsky, P;Oko, RJ
核周膜（PT）是精子头部的重要附属结构，但其生物起源尚不清楚。为了了解精子发生过程中PT衍生的体细胞组蛋白的发育起源，我们使用了亲和纯化的抗体对体细胞类型的历史H3，H2 B，H2 A和H4探测牛睾丸组织使用三种不同的免疫定位技术。虽然在拉长精子细胞核中检测不到，但免疫过氧化物酶光学显微镜显示，所有四个体细胞的历史仍然与牛精子发生的14个步骤中的第11至14步精子细胞的尾状头区域相关。免疫金电子显微镜证实了本地化的体细胞历史上的两个非核结构，即短暂的manchette微管的步骤-9到步骤-11精子细胞和发展的顶体后鞘的步骤-13和-14精子细胞。免疫荧光显示，体细胞组蛋白免疫反应性在发展中的顶体后鞘，和抗β-微管蛋白装饰的第12步精子细胞的柄状微管。通过电子显微镜和免疫荧光检测到圆形精子细胞核的基础上的局灶性抗核孔复合物标记，发生在精子细胞伸长过程中的核蛋白过渡期之前。这表明，如果核组蛋白输出之前，他们的降解，这一过程只能发生在这一地区，从而质疑所提出的作用的manchette在核质运输。顶体后鞘形成过程中的体细胞组蛋白免疫组化检测支持在PT组装的manchette的作用，这意味着一些PT组件有起源于远端精子细胞的细胞质。此外，这些研究结果表明，体细胞的历史从头合成后期精子PT大会。
The perinuclear theca (PT) is an important accessory structure of the sperm head, yet its biogenesis is not well defined. To understand the developmental origins of PT-derived somatic histones during spermiogenesis, we used affinity-purified antibodies against somatic-type histories H3, H2B, H2A, and H4 to probe bovine testicular tissue using three different immunolocalization techniques. While undetectable in elongating spermatid nuclei, immunoperoxidase light microscopy showed all four somatic histories remained associated to the caudal head region of spermatids from steps 11 to 14 of the 14 steps in bovine spermiogenesis. Immunogold electron microscopy confirmed the localization of somatic histories on two nonnuclear structures, namely transient manchette microtubules of step-9 to step-11 spermatids and the developing postacrosomal sheath of step-13 and -14 spermatids. Immunofluorescence demonstrated somatic histone immunoreactivity in the developing postacrosomal sheath, and on anti-beta-tubulin decorated manchette microtubules of step-12 spermatids. Focal antinuclear pore complex labeling on the base of round spermatid nuclei was detected by electron microscopy and immunofluorescence, occurring before the nucleoprotein transition period during spermatid elongation. This indicated that, if nuclear histone export precedes their degradation, this process could only occur in this region, thereby questioning the proposed role of the manchette in nucleocytoplasmic trafficking. Somatic histone immunodetection on the manchette during postacrosomal sheath formation supports a role for the manchette in PT assembly, signifying that some PT components have origins in the distal spermatid cytoplasm. Furthermore, these findings suggest that somatic histories are de novo synthesized in late spermiogenesis for PT assembly.