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增殖细胞核抗原的免疫组织化学定位作为研究啮齿动物和灵长类动物睾丸细胞增殖的一种工具。

Immunohistochemical localization of proliferating cell nuclear antigen as a tool to study cell proliferation in rodent and primate testes.

作者信息

Schlatt S, Weinbauer G F

机构信息

Institute of Reproductive Medicine, University, Münster, Germany.

出版信息

Int J Androl. 1994 Aug;17(4):214-22. doi: 10.1111/j.1365-2605.1994.tb01245.x.

Abstract

The immunohistochemical localization of proliferating cell nuclear antigen (PCNA) has become a widely used method for the detection of proliferating cells in cancerous tissues. PCNA expression is maximal around the S phase of the cell cycle. This study has evaluated the applicability of PCNA localization for the analysis of germ cell proliferation in rats, Djungarian hamsters, rhesus and cynomolgus monkeys and men, using Bouin's-fixed, paraplast-embedded tissue. In addition, involuted testes from photoinhibited hamsters, testes from immature rhesus monkeys and from GnRH antagonist-treated rats and cynomolgus monkeys were included. Monoclonal mouse anti-PCNA antibody (clone: PC10) was used for detection of the antigen. Visualization was performed by immunogold-silver staining or avidin-biotin staining. PCNA labelling was confined to the nuclei of spermatogonia and early spermatocytes within the seminiferous epithelium of all species. The distribution of PCNA among the different types of A-spermatogonia in primates is in good agreement with the previously described proliferation pattern of these cells. No staining was observed in resting A-dark spermatogonia, while differentiating A-pale spermatogonia were positive at distinct stages of the seminiferous epithelium cycle. In the rodent species the pattern of labelled A-spermatogonia was stage-specific, but agreed only partly with the previously described pattern of mitotic figures of A-spermatogonia. Hormonal withdrawal induced a decrease in the number of PCNA-positive cells in adult rats, hamsters and monkeys. In immature testes from rhesus monkeys positive staining was present in spermatogonia but also in some Sertoli cells, indicating proliferative activity of Sertoli cells in the prepubertal stage.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

增殖细胞核抗原(PCNA)的免疫组化定位已成为检测癌组织中增殖细胞的一种广泛应用的方法。PCNA表达在细胞周期的S期左右达到最大值。本研究评估了使用Bouin固定、石蜡包埋组织,PCNA定位在大鼠、黑线仓鼠、恒河猴、食蟹猴和人类生殖细胞增殖分析中的适用性。此外,还纳入了光抑制仓鼠的退化睾丸、未成熟恒河猴的睾丸以及GnRH拮抗剂处理的大鼠和食蟹猴的睾丸。使用单克隆小鼠抗PCNA抗体(克隆:PC10)检测抗原。通过免疫金银染色或抗生物素蛋白-生物素染色进行可视化。PCNA标记局限于所有物种生精上皮内精原细胞和早期精母细胞的细胞核。灵长类动物中不同类型A精原细胞之间PCNA的分布与这些细胞先前描述的增殖模式高度一致。静止的A暗精原细胞未观察到染色,而分化中的A淡精原细胞在生精上皮周期的不同阶段呈阳性。在啮齿动物物种中,标记的A精原细胞模式具有阶段特异性,但仅部分与先前描述的A精原细胞有丝分裂图像模式一致。激素撤退导致成年大鼠、仓鼠和猴子中PCNA阳性细胞数量减少。在恒河猴未成熟睾丸中,精原细胞以及一些支持细胞中存在阳性染色,表明支持细胞在青春期前阶段具有增殖活性。(摘要截断于250字)

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