Regulation of GDF9 and CDKN1B expression in Tibetan sheep testes during different stages of maturity

IF 1 4区生物学 Q4 DEVELOPMENTAL BIOLOGY

Gene Expression Patterns Pub Date : 2022-03-01 DOI:10.1016/j.gep.2021.119218

Huihui Wang , Xia Wang , Taotao Li , Xuejiao An , De'en Yin , Nana Chen , Youji Ma

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引用次数: 2

Abstract

Normal spermatogenesis is heavily dependent on the balance of germ cell proliferation, differentiation and apoptosis. Growth differentiation factor 9 (GDF9) and cyclin-dependent kinase inhibitor 1 B (CDKN1B) are strongly associated with cell cycle transition from G0/G1 to S and G2/M phase and hence regulating the growth and development of testicular germ cells and somatic cells. The current study was aimed at seeking out scientific evidence to determine if GDF9 and CDKN1B gene expression functions in the development of Tibetan sheep testes. To this end, developmental testes were derived from three-month-old (pre-puberty), one-year-old (sexual maturity), and three-year-old (adult) Tibetan sheep and then the expression and localization patterns of GDF9 and CDKN1B in these testes were evaluated using quantitative real-time PCR (qRT-PCR), Western blot and immunofluorescence. qRT-PCR and Western blot results showed that GDF9 and CDKN1B were detected in the testes throughout the different developmental stages. The abundance of GDF9 mRNA and protein in the testes of one- and three-year-old Tibetan sheep were higher than that in the testes of three-month-old Tibetan sheep; the mRNA and protein abundance of the CDKN1B gene in three-month-old Tibetan sheep testes were higher than that in the testes of the one-and three-year-old sheep. Moreover, immunofluorescence results suggested that the GDF9 protein was expressed in spermatogonia and Leydig cells, and that the CDKN1B protein was localized mainly in Leydig cells with some in the seminiferous epithelium throughout developmental stages. This indicated a novel role of the GDF9 and CDKN1B genes in Leydig cell development over and above their known roles in germ cell development. These findings have significant implications for our understanding of the molecular mechanisms of GDF9 and CDKN1B genes in Tibetan sheep spermatogenesis.

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藏羊不同成熟期睾丸GDF9和CDKN1B表达的调控

正常的精子发生在很大程度上依赖于生殖细胞增殖、分化和凋亡的平衡。生长分化因子9 (GDF9)和细胞周期蛋白依赖性激酶抑制剂1 B (CDKN1B)与细胞周期从G0/G1期向S期和G2/M期过渡密切相关，从而调节睾丸生殖细胞和体细胞的生长发育。目前的研究旨在寻找科学证据，以确定GDF9和CDKN1B基因表达是否在藏羊睾丸发育中起作用。为此，选取3月龄(青春期前)、1月龄(性成熟)和3岁(成年)藏羊的发育睾丸，采用实时荧光定量PCR (qRT-PCR)、Western blot和免疫荧光技术检测GDF9和CDKN1B在这些睾丸中的表达和定位模式。qRT-PCR和Western blot结果显示，在不同发育阶段的睾丸中均检测到GDF9和CDKN1B。1岁和3岁藏羊睾丸中GDF9 mRNA和蛋白的丰度高于3月龄藏羊;3月龄藏羊睾丸CDKN1B基因mRNA和蛋白丰度均高于1岁和3岁藏羊睾丸。此外，免疫荧光结果表明，GDF9蛋白在精原细胞和间质细胞中表达，CDKN1B蛋白在整个发育阶段主要定位于间质细胞，部分位于精原上皮。这表明GDF9和CDKN1B基因在间质细胞发育中的新作用超出了它们在生殖细胞发育中的已知作用。这些发现对我们理解GDF9和CDKN1B基因在藏羊精子发生中的分子机制具有重要意义。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Gene Expression Patterns 生物-发育生物学

CiteScore

2.30

自引率

0.00%

发文量

审稿时长

35 days

期刊介绍： Gene Expression Patterns is devoted to the rapid publication of high quality studies of gene expression in development. Studies using cell culture are also suitable if clearly relevant to development, e.g., analysis of key regulatory genes or of gene sets in the maintenance or differentiation of stem cells. Key areas of interest include: -In-situ studies such as expression patterns of important or interesting genes at all levels, including transcription and protein expression -Temporal studies of large gene sets during development -Transgenic studies to study cell lineage in tissue formation