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Identification of a novel testis-specific gene in mice and its potential roles in spermatogenesis.

ABSTRACT: AIM: Identification of a novel gene in mouse testis and its relation to spermatogenesis. METHODS: Genes expressed during different developmental stages of the mouse testis were screened by DNA microarray. The results of chip analysis were authenticated by reverse transcription-polymerase chain reaction (RT-PCR) technique, as well as the tissue distribution of the selected genes. The characteristics of the selected genes were analyzed by bioinformatics tools. RESULTS: A novel gene, TSC77, was identified and located at the mouse chromosome 2G1. The full cDNA length of TSC77 was 2280 bp, with a 2046 bp open reading frame encoding a 681 amino acids protein with a predicted molecular weight of 77.17 kDa. The results of subcellular localization of GFP-TSC77 fusion protein indicated TSC77 protein was located in the nucleus of Cos-7 cells. The analysis of multiple amino acid sequence alignment showed that TSC77 protein was highly homologous with the human CAI40813 (C2orf26 gene, 76%), and rat XP_230651 (77%). Three putative domains including nicotine amide dinucleotide phosphate (NAD(P))-nitrite reductase (NirB) domain, uncharacterized NAD flavin adenine dinucleotide (FAD)-dependent dehydrogenases (HcaD) domain, and nicotinamide adenine dinucleotide (NADH) dehydrogenase (Ndh) domain were predicted at the protein site 168-309, 187-245, 181-216, respectively. Gene expression anlysis showed that the mouse TSC77 is preferentially expressed in the mouse testis and its expression increased gradually from the day 9 to day 21. CONCLUSION: Based on its expression during mouse development, TSC77 may play an important role during mouse spermatogenesis.

SUBMITTER: Tang A

PROVIDER: S-EPMC2080497 | biostudies-literature | 2007 Feb

REPOSITORIES: biostudies-literature

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Identification of a novel testis-specific gene in mice and its potential roles in spermatogenesis.

Tang Aifa A Yu Zhendong Z Gui Yaoting Y Zhu Hui H Long Yun Y Cai Zhiming Z

Croatian medical journal 20070201 1

<h4>Aim</h4>Identification of a novel gene in mouse testis and its relation to spermatogenesis.<h4>Methods</h4>Genes expressed during different developmental stages of the mouse testis were screened by DNA microarray. The results of chip analysis were authenticated by reverse transcription-polymerase chain reaction (RT-PCR) technique, as well as the tissue distribution of the selected genes. The characteristics of the selected genes were analyzed by bioinformatics tools.<h4>Results</h4>A novel g ...[more]

PMID: 17309138

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Project description:Spermatogenesis is the highly orchestrated process involving expression of a series of testicular genes. Testis-enriched genes are critical for cellular processes during spermatogenesis whose disruption leads to impaired spermatogenesis and male infertility. Nevertheless, among poorly investigated testicular genes are the mouse Samd4a and human SAMD4A which were identified in the current study as novel testis-enriched genes through transcriptomic analyses. In particular, as orthologous alternative splicing isoforms, mouse Samd4a E-form and human SAMD4AC-form containing the SAM domain were specific to testes. Western blot analyses revealed that the murine SAMD4AE-form was predominantly found in the testis. Analyses on GEO2R and single-cell RNA-seq datasets revealed that the Samd4a/SAMD4A expression was enriched in spermatids among various types of cells in adult testes. To investigate in vivo functions of Samd4a, Samd4a knockout mice were generated using the CRISPR/Cas9 system. The Samd4a deficiency resulted in lower testis weight, absence of elongated spermatids, and an increased number of apoptotic cells. Profiling of gene expression in human testis samples revealed that the SAMD4A expression was comparable between obstructive azoospermia patients and normal controls, but significantly lowered in nonobstructive azoospermia (NOA) patients. Among three subgroups of NOA, pre-meiotic arrest (NOA-pre), meiotic arrest (NOA-mei), and post-meiotic arrest (NOA-post), expression level of SAMD4A was higher in the NOA-post than the NOA-mei, but there was no difference between the NOA-pre and NOA-mei. The current studies demonstrated spermatid stage-specific expression of Samd4a/SAMD4A, and impairment of the late stages of spermatogenesis by disruption of the mouse Samd4a gene. These data suggest that Samd4a/SAMD4A plays an essential role in normal spermatogenesis, and SAMD4A, as a spermatid specific marker, can be used for subcategorizing NOA patients. Further understanding the molecular role of SAMD4A will advance our knowledge on genetic regulations in male infertility.

Dataset Information

Identification of a novel testis-specific gene in mice and its potential roles in spermatogenesis.

Publications

Identification of a novel testis-specific gene in mice and its potential roles in spermatogenesis.

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