Immunolocalization of TAR DNA-binding protein of 43 kDa (TDP-43) in mouse seminiferous epithelium.
Immunolocalization of TAR DNA-binding protein of 43 kDa (TDP-43) in mouse seminiferous epithelium.
复制标题
小鼠生精上皮中 43-kDa 的 TAR DNA 结合蛋白 (TDP-43) 的免疫定位。
DOI：
10.1002/mrd.22851
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发表时间：
2017
影响因子：
2.5
通讯作者：
Reddi,PrabhakaraP
中科院分区：
文献类型：
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作者：
Osuru,HariPrasad;Pramoonjago,Patcharin;Abhyankar,MayureshM;Swanson,Eric;Roker,LaToyaAnn;Cathro,Helen;Reddi,PrabhakaraP
TAR DNA结合蛋白43 kDa（TDP-43）是一种进化上保守、普遍表达的多功能DNA/RNA结合蛋白，在基因转录、mRNA剪接、稳定性、转运、微RNA生物合成和转座子抑制中发挥作用。睾丸和精子中TDP-43的异常表达最近被证明与男性不育有关，这突出了更好地了解睾丸中TDP-43表达的必要性。我们之前从小鼠睾丸cDNA文库中克隆了TDP-43，并表明它作为转录抑制因子发挥作用，并在精子发生过程中调节编码顶体蛋白SP-10的Acrv 1基因的精确时空表达。在这里，我们使用识别TDP-43的氨基和羧基末端的四种单独的抗体对小鼠睾丸进行免疫印迹和免疫组织化学。TDP-43存在于生殖细胞以及支持细胞的细胞核中。TDP-43表达始于B型/中间型精原细胞，在细线期前精母细胞中达到峰值，在细线期和偶线期精母细胞中检测不到。粗线期精母细胞和早期圆形精子细胞再次表达TDP-43，但其丰度随后在精子细胞中减少（在步骤5-8）。有趣的是，四种抗体中的两种在第9-10步显示出精子细胞中的TDP-43表达，这与组蛋白向鱼精蛋白转变的初始阶段相一致。研究中观察到的免疫反应性模式表明，TDP-43在精子发生的不同阶段呈现不同的构象状态。TDP-43病理学已在神经退行性疾病的背景下进行了广泛研究;其在精子发生中的作用需要进一步详细研究TDP-43在男性不育中的作用。
TAR DNA‐binding protein of 43 kDa (TDP‐43) is an evolutionarily conserved, ubiquitously expressed, multi‐functional DNA/RNA‐binding protein with roles in gene transcription, mRNA splicing, stability, transport, micro RNA biogenesis, and suppression of transposons. Aberrant expression of TDP‐43 in testis and sperm was recently shown to be associated with male infertility, which highlights the need to understand better the expression of TDP‐43 in the testis. We previously cloned TDP‐43 from a mouse testis cDNA library, and showed that it functions as a transcriptional repressor and regulates the precise spatiotemporal expression of theAcrv1gene, which encodes the acrosomal protein SP‐10, during spermatogenesis. Here, we performed immunoblotting and immunohistochemistry of the mouse testis using four separate antibodies recognizing the amino and carboxyl termini of TDP‐43. TDP‐43 is present in the nuclei of germ cells as well as Sertoli cells. TDP‐43 expression begins in type B/intermediate spermatogonia, peaks in preleptotene spermatocytes, and becomes undetectable in leptotene and zygotene spermatocytes. Pachytene spermatocytes and early round spermatids again express TDP‐43, but its abundance diminishes later in spermatids (at steps 5–8). Interestingly, two of the four antibodies showed TDP‐43 expression in spermatids at steps 9–10, which coincides with the initial phase of the histone‐to‐protamine transition. Immunoreactivity patterns observed in the study suggest that TDP‐43 assumes different conformational states at different stages of spermatogenesis. TDP‐43 pathology has been extensively studied in the context of neurodegenerative diseases; its role in spermatogenesis warrants further detailed investigation of the involvement of TDP‐43 in male infertility.