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在不同发育阶段的大鼠睾丸中 attractin 的表达和分布的变化。

Changes in expression and distribution of attractin in the testes of rats at different developmental stages.

机构信息

Department of Andrology, Jinling Hospital, Nanjing 210002, P.R. China.

出版信息

Int J Mol Med. 2013 Sep;32(3):599-606. doi: 10.3892/ijmm.2013.1423. Epub 2013 Jun 21.

Abstract

Attractin (Atrn), an autosomal recessive gene, is widely expressed in the body and displays multiple physiological and pathological functions in different types of tissues. The objective of this study was to localize Atrn protein and mRNA in the testis and epididymis of rats at different stages of maturation. Testis and epididymidis samples were obtained from the following 5 groups of Sprague Dawley (SD) rats in different developmental stages: newborn (8 h after birth), prepubertal (5 days), pubertal (20 days), postpubertal (50 days) and mature (70 days). Tissues were fixed and prepared for indirect immunofluorescence, immunohistochemistry, in situ hybridization, confocal laser scanning microscopy and western blot assays. A polyclonal antiserum against mouse Atrn and oligonucleotide riboprobes were used in the above assays. At the different stages of maturation, Atrn protein and mRNA were both widely expressed in the rat testis, including Leydig cells, primitive spermatogonia, primary spermatocytes, spermatids, Sertoli and peritubular myoid cells. Staining of the Atrn protein was mainly located on the cell membrane and in the cell cytoplasm while Atrn mRNA was distributed in both the nucleus and cytoplasm. No immunopositive staining was detected in spermatozoa and epididymides. In the epididymis, comprised of the caput, corpus and cauda, there was no definitive immunopositive staining within the efferent ductules or epididymal ducts. Taken together, Atrn protein and mRNA are both expressed widely in the rat testis at different stages of maturation, which suggests that Atrn protein is involved and plays an important role in the development of the reproductive system. In addition, the rat testis has the ability to synthesize Atrn protein throughout sexual development.

摘要

Attractin (Atrn),一种常染色体隐性基因,在体内广泛表达,在不同类型的组织中具有多种生理和病理功能。本研究旨在定位 Atrn 蛋白和 mRNA 在不同成熟阶段大鼠睾丸和附睾中的表达。从以下五个不同发育阶段的 Sprague Dawley (SD) 大鼠中获得睾丸和附睾样本:新生(出生后 8 小时)、青春期前(5 天)、青春期(20 天)、青春期后(50 天)和成熟(70 天)。组织固定并准备进行间接免疫荧光、免疫组织化学、原位杂交、共聚焦激光扫描显微镜和 Western blot 检测。使用针对小鼠 Atrn 的多克隆抗血清和寡核苷酸探针进行上述检测。在不同的成熟阶段,Atrn 蛋白和 mRNA 在大鼠睾丸中广泛表达,包括 Leydig 细胞、原始精原细胞、初级精母细胞、精子细胞、Sertoli 和小管周肌样细胞。Atrn 蛋白染色主要位于细胞膜和细胞质中,而 Atrn mRNA 分布于细胞核和细胞质中。在精子和附睾中未检测到免疫阳性染色。在由头部、体部和尾部组成的附睾中,在输出小管或附睾管内没有明确的免疫阳性染色。综上所述,Atrn 蛋白和 mRNA 在不同成熟阶段的大鼠睾丸中广泛表达,表明 Atrn 蛋白参与并在生殖系统发育中发挥重要作用。此外,大鼠睾丸在性发育过程中具有合成 Atrn 蛋白的能力。

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