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斑马鱼睾丸特异性基因10(tsga10)的鉴定与表达分析。

Identification and expression analysis of zebrafish testis-specific gene 10 (tsga10).

作者信息

Asghari-Givehchi Shohreh, Hossein-Modarressi Mohammad

机构信息

Department of Medical Genetics, School of Medicine, International Campus, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Int J Dev Biol. 2019;63(11-12):623-629. doi: 10.1387/ijdb.190053mm.

DOI:10.1387/ijdb.190053mm
PMID:32149372
Abstract

Several clinical studies suggest that testis-specific gene antigen 10 (TSGA10) is a cancer-testis antigen with a discernible expression pattern in the testis. Recent studies have highlighted that TSGA10 overexpression in HeLa cells impairs the transcriptional activity of hypoxia-inducible factor alpha (HIF-1α) and inhibits angiogenesis. In this study, we used the zebrafish as a powerful model organism to identify and characterize the orthologue of TSGA10. We analyzed the gene expression pattern by RT-PCR and whole mount in situ hybridization and overexpressed the tsga10 protein by mRNA microinjection. Our results revealed that during early development, tsga10 expression is enriched, but gradually subsides between 0 and 72 hours post fertilization (hpf). There was no detectable transcript at the larval stages. In adult fish, we found high expression levels of tsga10 in the testis and unfertilized egg and low levels of gene expression in the brain, eyes and muscle. Overexpression of tsga10, using tsga10 mRNA microinjection into one-cell stage embryos, resulted in angiogenic and morphological defects at 24 and 48 hpf. This study clarified the expression pattern of tsga10 in different developmental stages and adult tissues, suggesting that tsga10 may have a related biological role in different cell types and tissues. Our results indicate that tsga10 mRNA at embryonic stages is maternally deposited, indicating a transient functional role during embryogenesis. Our findings suggest that tsga10 is a human orthologous gene relevant for future studies to elucidate its mechanism of action in angiogenesis.

摘要

多项临床研究表明,睾丸特异性基因抗原10(TSGA10)是一种癌症-睾丸抗原,在睾丸中具有明显的表达模式。最近的研究强调,HeLa细胞中TSGA10的过表达会损害缺氧诱导因子α(HIF-1α)的转录活性并抑制血管生成。在本研究中,我们将斑马鱼作为一种强大的模式生物,以鉴定和表征TSGA10的直系同源物。我们通过逆转录聚合酶链反应(RT-PCR)和整体原位杂交分析基因表达模式,并通过mRNA显微注射使tsga10蛋白过表达。我们的结果显示,在早期发育过程中,tsga10表达丰富,但在受精后0至72小时(hpf)之间逐渐减弱。在幼体阶段未检测到转录本。在成年鱼中,我们发现tsga10在睾丸和未受精卵中高表达,而在脑、眼和肌肉中基因表达水平较低。将tsga10 mRNA显微注射到单细胞期胚胎中过表达tsga10,在24和48 hpf时导致血管生成和形态缺陷。本研究阐明了tsga10在不同发育阶段和成年组织中的表达模式表明tsga10可能在不同细胞类型和组织中具有相关生物学作用。我们的结果表明,胚胎阶段的tsga10 mRNA是母源沉积的,表明在胚胎发生过程中具有短暂的功能作用。我们的研究结果表明,tsga10是一个人类直系同源基因,与未来阐明其血管生成作用机制的研究相关。

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