Abstract

The identification and characterization of germ cell-specific genes are essential if we hope to comprehensively understand the mechanisms of spermatogenesis and fertilization. Here, we searched the mouse UniGene databases and identified 13 novel genes as being putatively testis-specific or -predominant. Our in silico and in vitro analyses revealed that the expressions of these genes are testis- and germ cell-specific, and that they are regulated in a stage-specific manner during spermatogenesis. We generated antibodies against the proteins encoded by seven of the genes to facilitate their characterization in male germ cells. Immunoblotting and immunofluorescence analyses revealed that one of these proteins was expressed only in testicular germ cells, three were expressed in both testicular germ cells and testicular sperm, and the remaining three were expressed in sperm of the testicular stages and in mature sperm from the epididymis. Further analysis of the latter three proteins showed that they were all associated with cytoskeletal structures in the sperm flagellum. Among them, MORN5, which is predicted to contain three MORN motifs, is conserved between mouse and human sperm. In conclusion, we herein identify 13 authentic genes with male germ cell-specific expression, and provide comprehensive information about these genes and their encoded products. Our finding will facilitate future investigations into the functional roles of these novel genes in spermatogenesis and sperm functions.

Highlights

  • During spermatogenesis, germ cells are processed from primordial germ cells (PGCs) to mature sperm [1, 2]

  • To select for genes that are exclusively transcribed in testis, we calculated testis specificity using information from the mouse UniGene database (Build #193) [9], which includes the expressed sequence tag (EST) expression profile of a given gene in particular tissues of mice at a specific age and/or health status, as expressed in TPM

  • We found that Mm.73222/TEX33 was present only in testicular cells; Mm.258841/ FAM71E1, Mm.131623/TMCO5, and Mm.272519/1700013G24Rik existed in testicular cells and sperm but not in mature sperm; and Mm.276332/MORN5, Mm.271255/1700013F07Rik, and Mm.159422/4930505A04Rik were present in all stages

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Summary

Introduction

Germ cells are processed from primordial germ cells (PGCs) to mature sperm [1, 2]. The tightly regulated nature of this process, which occurs in the seminiferous tubules of testes, indicates that a highly organized network of genes is expressed in male germ cells during their development [3]. An intrinsic program determines which genes are utilized and when they are expressed in germ cells, and an interactive regulation between germ cells and somatic cells is necessary for germ cell proliferation and progression. Extrinsic influences, including steroid and peptide hormones, control this interactive regulation. The intrinsic genetic program is central to the development and fertilization of male germ cells.

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