A Novel Testicular Haploid Expressed Gene (THEG) Involved in Mouse Spermatid-Sertoli Cell Interaction1

Nayernia, Karim; Mering, M. H. P. Von; Kraszucka, K.; Burfeind, Peter; Wehrend, Axel; Köhler, Martin; Schmid, Michael; Engel, Wolfgang

doi:10.1095/biolreprod60.6.1488

Cited by 21 publications

(33 citation statements)

References 36 publications

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“…2A). Included in this cluster were numerous mRNAs implicated in testicular and/or sperm function, including Theg (Nayernia et al 1999;Yanaka et al 2000), Tcp10 (Schimenti et al 1988), Man2a2 (Akama et al 2002), Adam1b/ Ftna (Cho et al 1997), Adam24 (Zhu et al 1999), Clgn (Ikawa et al 1997), and Dnahc8 (Samant et al 2002). The next largest cluster contained 12 ncRNAs and 19 mRNAs, all of which were highly expressed in the central nervous system (Supplemental Fig.…”

Section: Tissue Profiling Of Putative Ncrnasmentioning

confidence: 99%

Experimental validation of the regulated expression of large numbers of non-coding RNAs from the mouse genome

Ravasi¹,

Suzuki²,

Pang³

et al. 2005

Genome Res.

473

364

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Recent large-scale analyses of mainly full-length cDNA libraries generated from a variety of mouse tissues indicated that almost half of all representative cloned sequences did not contain an apparent protein-coding sequence, and were putatively derived from non-protein-coding RNA (ncRNA) genes. However, many of these clones were singletons and the majority were unspliced, raising the possibility that they may be derived from genomic DNA or unprocessed pre-mRNA contamination during library construction, or alternatively represent nonspecific "transcriptional noise." Here we show, using reverse transcriptase-dependent PCR, microarray, and Northern blot analyses, that many of these clones were derived from genuine transcripts of unknown function whose expression appears to be regulated. The ncRNA transcripts have larger exons and fewer introns than protein-coding transcripts. Analysis of the genomic landscape around these sequences indicates that some cDNA clones were produced not from terminal poly(A) tracts but internal priming sites within longer transcripts, only a minority of which is encompassed by known genes. A significant proportion of these transcripts exhibit tissue-specific expression patterns, as well as dynamic changes in their expression in macrophages following lipopolysaccharide stimulation. Taken together, the data provide strong support for the conclusion that ncRNAs are an important, regulated component of the mammalian transcriptome.[Supplemental material is available online at www.genome.org. The microarray data from this study have been submitted to the Gene Expression Omnibus under accession nos. GSD275 and GSE3098.] In recent years there have been increasing reports of functional non-protein-coding RNAs (ncRNAs) that are involved or implicated in developmental, tissue-specific, and disease processes, including X-chromosome dosage compensation, germ cell development and embryogenesis, neural and immune cell development, kidney and testis development, B-cell neoplasia, lung cancer, prostate cancer, cartilage-hair hypoplasia, spinocerebellar ataxia type 8, DiGeorge syndrome, autism, and schizophrenia (see Pang et al. 2005). Many putative ncRNAs are alternatively spliced and/or polyadenylated (Sutherland et al. 1996;Tam et al. 1997;Bussemakers et al. 1999;Raho et al. 2000;Charlier et al. 2001;Wolf et al. 2001). Smaller ncRNAs, termed microRNAs, have also been shown to be involved in developmental processes in both plants and animals, as well as implicated in disease (Carrington and Ambros 2003;Mattick and Makunin 2005). Recent evidence suggests that these microRNAs are derived from the introns of capped and polyadenylated protein-coding transcripts as well as the exons and introns of non-protein-coding transcripts, many of which are derived from "intergenic" regions (Cai et al. 2004;Rodriguez et al. 2004;Seitz et al. 2004;Mattick and Makunin 2005;Ying and Lin 2005). In addition, many complex genetic phenomena, including cosuppression, imprinting, methylation, and gene silencing (see Mattick...

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Section: Tissue Profiling Of Putative Ncrnasmentioning

confidence: 99%

Experimental validation of the regulated expression of large numbers of non-coding RNAs from the mouse genome

Ravasi¹,

Suzuki²,

Pang³

et al. 2005

Genome Res.

473

364

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“…In the process of spermatogenesis, spermatid-Sertoli cell interaction and the normal hormone production from Leydig cell are essential [23][24][25]. Endocrine disruptors, also called xenobiotic substances, can disturb the normal sexual hormonal system of the reproductive system of mammals.…”

Section: Discussionmentioning

confidence: 99%

A Novel Spermatogenesis Related Factor-2 (SRF-2) Gene Expression Affected by TCDD Treatment

et al. 2005

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Abstract. We have cloned a gene which is specifically expressed at the stage of sexual maturation in the rat testis by means of differential display, and have named it spermatogenesis-related factor-2 (SRF-2). Testicular expression was first detected at 5 weeks of age, and its level of the expression increased up to 7 weeks, and was maintained even at 63 weeks. Its cDNA was 2,789 bp in length and encoded an open reading frame of 718 amino acids. This gene was mainly expressed in the spermatocyte, judging from the result of in situ hybridization. The hypothetical gene product had a motif highly homologous with RabGAP/TBC protein. Taken together, this gene is considered to have some important functions for meiosis. The gene expression was significantly decreased by treatment with TCDD, a candidate endocrine disruptor, when administered to male rats of the nursling period. Body weight and testis weight were decreased by the treatment, but even then the sperm concentration in cauda epididymis was not changed significantly. SRF-2 gene may be a promising biomarker to construct a detection system of uncertain endocrine disruptors.

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“…For immunostaining of mouse THEG proteins on mouse testis sections a polyclonal antibody was generated (Eurogentec) which is directed against two different epitopes (peptide 1: amino acid position 44 to 58; peptide 2: amino acid position 183 to 197), originated from the deduced mouse THEG amino acid sequence (Nayernia et al, 1999). The affinity purifications of the polyclonal antibody against THEG were performed using HiTrap TM NHSactivated sepharose columns according to the manufacturer's instructions (Amersham Pharmacia Biotech).…”

Section: Immunocytochemical Analysis and Construction Of Gfp-theg Fusmentioning

confidence: 99%

“…In previous studies, our group used the immortalized mouse Sertoli cell line 15P-1, which supports the multiplication of diploid spermatogenic precursors and their transit through meiosis in coculture (Rassoulzadegan et al, 1993). The mRNA differential display technique (Liang et al, 1992) was used for analysis of changes in gene expression while coculturing 15P-1 cells and spermatids (Nayernia et al, 1999). These studies enabled us to identify a novel mouse gene named Theg (Nayernia et al, 1999) that is expressed in spermatids.…”

Section: Introductionmentioning

confidence: 99%

“…The mRNA differential display technique (Liang et al, 1992) was used for analysis of changes in gene expression while coculturing 15P-1 cells and spermatids (Nayernia et al, 1999). These studies enabled us to identify a novel mouse gene named Theg (Nayernia et al, 1999) that is expressed in spermatids. In the previous study it was demonstrated that the maintenance/induction of mouse THEG gene expression in spermatids requires their interaction with Sertoli cells.…”

Section: Introductionmentioning

confidence: 99%

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Alternative splicing, chromosome assignment and subcellular localization of the testicular haploid expressed gene (THEG)

Mannan

Lucke²,

Dixkens³

et al. 2000

Cytogenet Genome Res

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We have previously isolated and characterized the mouse Testicular Haploid Expressed Gene (Theg) that is specifically expressed in haploid germ cells. We now describe the molecular cloning and characterization of the human homologue (THEG) of mouse Theg. Expression studies by using both dot blot and Northern blot techniques revealed that human THEG is expressed specifically in the testis. Additionally, we found two alternatively spliced transcripts (THEG major and THEG minor) for THEG by using reverse transcription-polymerase chain reaction on human testicular RNA. Sequence analysis of these PCR products demonstrated that the smaller transcript (THEG minor) lacks 72 bp which was also observed for the mouse Theg. We have isolated the cDNAs of human THEG major and THEG minor, containing the complete open reading frames, which encode putative nuclear proteins of 379 amino acids and 355 amino acids, respectively. Database searches identified two genomic clones on chromosome 19 harboring the human THEG gene, which is approximately 14 kb pairs in size, contains eight exons, and comparison of the two cDNA sequences with the genomic sequence indicated that the smaller transcript lacks exon 3. Furthermore, we assigned the human THEG gene (THEG) to human chromosome 19ptel→ p13 by fluorescence in situ hybridization. Moreover, we detected mouse THEG protein prominently in the nucleus of round spermatids by using an antibody against THEG on both testicular sections and cellular suspensions. Additionally, the subcellular localization of mouse THEG was confirmed by a green fluorescent protein (GFP) fusion protein of mouse THEG which was found mainly in the nucleus of transfected NIH3T3 cells. These data suggest that both human and mouse THEG are specifically expressed in the nucleus of haploid male germ cells and are involved in the regulation of nuclear functions.

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A Novel Testicular Haploid Expressed Gene (THEG) Involved in Mouse Spermatid-Sertoli Cell Interaction1

Cited by 21 publications

References 36 publications

Experimental validation of the regulated expression of large numbers of non-coding RNAs from the mouse genome

Experimental validation of the regulated expression of large numbers of non-coding RNAs from the mouse genome

A Novel Spermatogenesis Related Factor-2 (SRF-2) Gene Expression Affected by TCDD Treatment

Alternative splicing, chromosome assignment and subcellular localization of the testicular haploid expressed gene (THEG)

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