Immunohistochemistry and immunocytochemistry analysis of PLZF and VASA in mice testis during spermatogenesis.

IF 1.5 4区生物学 Q4 CELL BIOLOGY Zygote Pub Date : 2023-06-01 DOI:10.1017/S0967199423000047

Mohammad Babatabar Darzi, Farkhondeh Nemati, Hossein Azizi, Abbasali Dehpour Jouybari

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引用次数: 1

Abstract

Spermatogonial stem cells (SSCs) are the basis of male spermatogenesis and fertility. SSCs are distinguished by their ability to self-renew and differentiate into spermatozoa throughout the male reproductive life and pass genetic information to the next generation. Immunohistochemistry (IHC), immunocytochemistry (ICC) and Fluidigm reverse transcriptase-polymerase chain reaction (RT-PCR) were used to analyze the expression of PLZF and VASA in mice testis tissue. In this experimental study, whereas undifferentiated spermatogonial cells sharply expressed PLZF, other types of germ cells located in the seminiferous tubule were negative for this marker. Conversely, the germ cells near the basal membrane of the seminiferous tubule showed VASA expression, whereas the undifferentiated germ cells located on the basal membrane were negative. The ICC analysis indicated higher expression of PLZF in the isolated undifferentiated cells compared with differentiated germ cells. Fluidigm real-time RT-PCR results demonstrated a significant expression (P < 0.05) of VASA in the SSCs compared with differentiated cells and also showed expression of PLZF in undifferentiated spermatogonia. These results clearly proved the role of PLZF as a specific marker for SSCs, and can be beneficial for advanced research on in vitro differentiation of SSCs to functional sperms.

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精子发生过程中小鼠睾丸PLZF和VASA的免疫组织化学和免疫细胞化学分析。

精原干细胞(ssc)是男性精子发生和生育的基础。ssc的特点是在整个雄性生殖周期中具有自我更新和分化为精子的能力，并将遗传信息传给下一代。采用免疫组织化学(IHC)、免疫细胞化学(ICC)和Fluidigm逆转录聚合酶链反应(RT-PCR)分析小鼠睾丸组织中PLZF和VASA的表达。在本实验研究中，未分化的精原细胞急剧表达PLZF，而位于精小管的其他类型的生殖细胞则不表达该标记。相反，靠近精小管基膜的生殖细胞表达VASA，而位于基膜的未分化生殖细胞表达VASA。ICC分析表明，PLZF在分离的未分化细胞中的表达高于已分化的生殖细胞。Fluidigm实时RT-PCR结果显示，与分化细胞相比，ssc中VASA的表达显著(P < 0.05)，未分化精原细胞中PLZF的表达也显著。这些结果清楚地证明了PLZF作为ssc特异性标记物的作用，有助于进一步研究ssc向功能精子的体外分化。

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来源期刊

Zygote 生物-发育生物学

CiteScore

1.70

自引率

5.90%

发文量

117

审稿时长

6-12 weeks

期刊介绍： An international journal dedicated to the rapid publication of original research in early embryology, Zygote covers interdisciplinary studies on gametogenesis through fertilization to gastrulation in animals and humans. The scope has been expanded to include clinical papers, molecular and developmental genetics. The editors will favour work describing fundamental processes in the cellular and molecular mechanisms of animal development, and, in particular, the identification of unifying principles in biology. Nonetheless, new technologies, review articles, debates and letters will become a prominent feature.