Tlapakova Tereza, Nguyen Thi Minh Xuan, Vegrichtova Marketa, Sidova Monika, Strnadova Karolina, Blahova Monika, Krylov Vladimir
Charles University in Prague, Faculty of Science, Vinicna 7, Prague 2 128 44, Czech Republic.
Charles University in Prague, Faculty of Science, Vinicna 7, Prague 2 128 44, Czech Republic Laboratory of Gene Expression, Institute of Biotechnology, Academy of Sciences of the Czech Republic, Videnska 1083, Prague 4 142 20, Czech Republic.
Biol Open. 2016 Sep 15;5(9):1275-82. doi: 10.1242/bio.019265.
The origin of somatic cell lineages during testicular development is controversial in mammals. Employing basal amphibian tetrapod Xenopus tropicalis we established a cell culture derived from testes of juvenile male. Expression analysis showed transcription of some pluripotency genes and Sertoli cell, peritubular myoid cell and mesenchymal cell markers. Transcription of germline-specific genes was downregulated. Immunocytochemistry revealed that a majority of cells express vimentin and co-express Sox9 and smooth muscle α-actin (Sma), indicating the existence of a common progenitor of Sertoli and peritubular myoid cell lineages. Microinjection of transgenic, red fluorescent protein (RFP)-positive somatic testicular cells into the peritoneal cavity of X. tropicalis tadpoles resulted in cell deposits in heart, pronephros and intestine, and later in a strong proliferation and formation of cell-to-cell net growing through the tadpole body. Immunohistochemistry analysis of transplanted tadpoles showed a strong expression of vimentin in RFP-positive cells. No co-localization of Sox9 and Sma signals was observed during the first three weeks indicating their dedifferentiation to migratory-active mesenchymal cells recently described in human testicular biopsies.
在哺乳动物中,睾丸发育过程中体细胞谱系的起源存在争议。利用基底两栖类四足动物热带爪蟾,我们建立了一种源自幼年雄性睾丸的细胞培养物。表达分析显示一些多能性基因以及支持细胞、睾丸肌样细胞和间充质细胞标志物的转录。生殖系特异性基因的转录被下调。免疫细胞化学显示大多数细胞表达波形蛋白,并共同表达Sox9和平滑肌α-肌动蛋白(Sma),表明存在支持细胞和睾丸肌样细胞谱系的共同祖细胞。将转基因红色荧光蛋白(RFP)阳性的体细胞睾丸细胞显微注射到热带爪蟾蝌蚪的腹腔中,导致细胞沉积在心脏、前肾和肠道中,随后在蝌蚪体内强烈增殖并形成细胞间网络。对移植蝌蚪的免疫组织化学分析显示RFP阳性细胞中波形蛋白强烈表达。在前三周未观察到Sox9和Sma信号的共定位,表明它们去分化为最近在人类睾丸活检中描述的具有迁移活性的间充质细胞。