Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “mouse testis”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 541 records · Page 30Linked to original sources
PubMed2005

Rat testis as a radiobiological in vivo model for radionuclides.

The radiobiological effect of intracellularly localised radionuclides emitting low energy electrons (Auger electrons) has received much attention. Most in vivo studies reported have been performed in the mouse testis. We have investigated the rat testis as an in vivo radiobiological model, with sperm-head survival, testis weight loss and also alteration in the blood plasma hormone levels of FSH and LH as radiobiological endpoints. Validation of the rat testis model was evaluated by using mean absorbed doses of up to 10 Gy from intratesticularly (i.t.) injected (111)In oxine or local X-ray irradiation. Biokinetics of the i.t. injected radionuclide was analysed by scintillation camera imaging and used in the absorbed dose estimation. By the analysis of the autoradiographs, the activity distribution was revealed. Cell fractionation showed (111)In to be mainly associated with the cell nuclei. External irradiations were monitored by thermoluminescence dosimeters. The sperm-head survival was the most sensitive radiobiological parameter correlated to the mean absorbed dose, with a D(37) of 2.3 Gy for (111)In oxine and 1.3 Gy for X rays. The levels of plasma pituitary gonadal hormones FSH and LH were elevated for absorbed doses >7.7 Gy. This investigation shows that the radiobiological model based on the rat testis has several advantages compared with the previously commonly used mouse testis model. The model is appropriate for further investigations of basic phenomena such as radiation geometry, intracellular kinetics and heterogeneity, crucial for an understanding of the biological effect of low-energy electrons.

Animals↗
PubMed2006

Organotypic culture, a powerful model for studying rat and mouse fetal testis development.

The key role of the fetal testis in the masculinization of genital organs has been known for a long time. More recently, the observed increases in male reproductive disorders has been postulated to be the result of changes in fetal and neonatal testis development in response to increasing environmental pollution. However, few tools are available for studying fetal testis development and the effects of physiological or toxic substances. We have developed an organ culture system in which rat fetal testis is grown on a filter floating on a synthetic medium containing no serum, hormones or biological factors. In this study, we have compared the long-term morpho-functional development of the various testicular cell types in this system with that observed in vivo and have extended this system to the mouse. Rat Leydig, Sertoli and germ cells and macrophages develop normally over a period of 1-2 weeks in this system. Fewer cells are produced than in vivo but the level of differentiated function is similar. Germ cells, which are difficult to culture in vitro, resume mitosis after a quiescent period, at the same time as in vivo. Similar results have been obtained with mouse fetuses, except that Leydig cells dedifferentiate in vitro if the testis is explanted after 13.5 days post conception. Testicular architecture and intercellular communication are sufficiently preserved for the development of the main fetal and neonatal testicular cell types in vitro with no added factors. Our floating-filter organotypic culture system in synthetic medium therefore allows the morpho-functional development of somatic and germ cells in fetal testis explants taken at all developmental stages in rat and at early stages in mouse. This method is potentially useful for studies of the effects of various factors, and of xenobiotics, in particular.

Animals↗
PubMed2005

Identification of a new transcript specifically expressed in mouse spermatocytes: mmrp2.

In an effort to examine the molecular basis of gametogenesis, we screened Riken cDNA database and the clone 4930481F22 that is expressed preponderantly in mouse testis was identified. In the course of the research, a new isoform of 4930481F22 clone was found, isolated from mouse testis and sequenced. It only lacks the 7th exon of 4930481F22 transcript. The new isoform only has 837 bp and encodes a putative 28.4 kDa protein. We investigated the expression pattern at the mRNA level by RT-PCR and in situ hybridization in testis. The new isoform was only expressed in the gonad, where it began to be detected at day 8 after birth. In situ hybridization proved that the new isoform mostly expressed in spermatocytes. The structure of the predicted protein and the expression pattern of the mRNA suggest that the new isoform could have an important role in meiosis. We temporarily named it mmrp 2 (Mouse Meiosis Related Protein 2).

Amino Acid Sequence↗
PubMed1995

Radiosensitivity of testicular cells in the fetal mouse.

The effects of prenatal X irradiation on postnatal development of the CBA/P mouse testis was studied. At days 14, 15 and 18 post coitus pregnant female mice were exposed to single doses of X rays ranging from 0.25-1.5 Gy. Higher doses resulted in extensive loss of fetal mice. In the male offspring, at days 3 and 31 post partum, the numbers of gonocytes, type A spermatogonia and Sertoli cells per testis were determined using the disector method. Furthermore, after irradiation at day 15 post coitus, the numbers of Leydig cells, mesenchymal cells, macrophages, myoid cells, lymphatic endothelial cells, endothelial cells and perivascular cells per testis were also determined at days 3 and 31 post partum. At day 3 post partum, the number of germ cells was decreased after irradiation at days 14 and 15 post coitus. A D0 value of 0.7 Gy was determined for the radiosensitivity of the gonocytes at day 14 post coitus. A D0 value of 0.8 Gy was determined for the gonocytes at day 15 post coitus which, however, seems to be less accurate. No accurate D0 value could be determined for the gonocytes at day 18 post coitus. At day 31 post partum, the repopulation of the seminiferous epithelium as well as testis weights and tubular diameters were more affected by irradiation with increasing age of the mice at the time of irradiation. The percentage of tubular cross sections showing spermatids decreased with increasing dose after irradiation at days 15 and 18 post coitus, but not after irradiation at day 14 post coitus. Furthermore, in tubular cross sections showing spermatids, exposure of testes to 1.25 and 1.5 Gy at day 18 post coitus resulted in significantly lower numbers of spermatids per cross section when compared to those testes exposed to the same doses at day 15 post coitus. This indicates that the radiosensitivity of the gonocytes increases with fetal age. Prenatal irradiation did not cause significant changes in the numbers per testis of the Sertoli cells or the interstitial cell types. The present results indicate that, in the fetal mouse testis, the spermatogonial stem cells are more sensitive to X irradiation than in the adult testis, while Sertoli cells and interstitial cells are relatively resistant.

Aging↗
PubMed1997

Tissue-specific subunit of the mouse cytosolic chaperonin-containing TCP-1.

We have cloned a novel Tcp-1-related mouse testis cDNA encoding a polypeptide of 531 amino acids which shares 81.2% identity with the zeta subunit of the mouse cytosolic chaperonin-containing TCP-1 (CCT). Immunoblot analysis of mouse testis CCT subunits separated by 2-dimensional gel electrophoresis indicates that this novel gene, Cctz-2, encodes a CCT subunit of Mr 57 000 and pI 7.1. Cctz-2 mRNA is detected only in testis whereas the other Cctz gene, Cctz-1, is expressed in all tissues investigated. The CCTzeta-2 subunit may have specific functions in the folding of testicular proteins and for interactions with testicular molecular chaperones.

Amino Acid Sequence↗
PubMed2003

Expression of the mouse Aven gene during spermatogenesis, analyzed by subtraction screening using Mvh-knockout mice.

As an attempt to investigate the function of the mouse Vasa-homolog (MVH) protein, a germ cell-specific RNA helicase, we have cloned genes that fail to be expressed in homozygous Mvh-knockout testes. One of the cDNA clones obtained by selective PCR subtraction cloning matched two cDNA sequences in the mouse testis cDNA database, and was highly homologous to Aven, a human gene regulating cell death. Sequences of the two mouse testis cDNAs in the database revealed that they share an 800 bp sequence but have quite different 5'-regions; the latter are derived from different exons by alternative use of transcription initiation sites. The shorter (mAven-S) and longer (mAven-L) transcripts encode 216 and 342 amino acids, respectively, and both isoforms are predominantly expressed in adult testes. However, mAven-L is not completely testis-specific and is expressed in testes during all developmental stages, whereas the expression of mAven-S is restricted to the testis and is barely detectable in Mvh-homozygous testes. In situ hybridization showed that mAven-S is expressed specifically in post-meiotic cells, mainly in round spermatids.

Amino Acid Sequence↗
PubMed2006

Identification and characteristics of a novel testis-specific gene, Tsc24, in human and mice.

Testis-specific genes are essential for the spermatogenesis in mammalian male reproduction. We have identified a novel gene, Tsc24, from the results of the Affymetrix Genechip analysis in the developmental stage of days 4, 9, 18, 35, 54 and 6 months of postnatal Balb/C mouse testis. The scaling signal intensities of Tsc24 in the six stages of mouse testis showed that the expression of Tsc24 was not detected on day 4, 9 or 18 but on day 35, 54 and 6 months. The full cDNA length of mouse Tsc24 was 899 bp, with a 624 bp open reading frame encoding a 207 amino acid protein with a predicted molecular weight of 23.997 kDa. The results of semi-quantitative RT-PCR showed that the expression of mouse Tsc24 can only be detected after the mouse was 35 d old and the expressing level increased gradually from day 35 to 6 months. Of the eight tissues (liver, spleen, heart, lung, brain, kidney, epididymis, and testis) examined in mice, and of the 12 tissues (liver, kidney, muscle, brain, spleen, adipose, lung, heart, epididymis, testis, ovary and uterus) examined in human, Tsc24 was exclusively expressed in the testis, but in none of the other studied tissues. The result of subcellular localization of GFP-Tsc24 fusion protein in Cos-7 cells supports that Tsc24 protein is expressed in nuclear. Our study should be a basis for function characterization of the Tsc24 gene, leading to the elucidation of the molecular events underlying mammalian male reproduction.

Age Factors↗
PubMed2004

Effect of neonatal exposure to diethylstilbestrol on testicular gene expression in adult mouse: comprehensive analysis with cDNA subtraction method.

Summary In utero or neonatal exposure to high levels of exogenous steroid hormones, such as the potent synthetic diethylstilbestrol (DES), incurs an increased risk of malfunctional male reproduction. In this study, we investigated whether neonatal exposure to DES induces the alteration of mRNA expression in adult mouse testis. Using a cDNA subtraction method, we isolated seven gene clones whose expression was changed in neonatally DES-treated mouse testis. Northern blot analysis revealed that five up-regulated genes (AF326230, AF356521, AK004975, AK006136 and BM237156) and two down-regulated genes (AK017044, AK017130) were predominantly expressed in testes of 8-week-old mice. Moreover, we confirmed that the expression of these seven genes was altered by neonatal DES-exposure using Northern blot analysis. Our results suggest that neonatal exposure to DES leads to the alteration of gene expression in the testis in the long term. These genes might be useful as biological markers of foetal or neonatal exposure to exogenous steroid hormones, such as DES.

Aging↗
PubMed2003

IFT20 links kinesin II with a mammalian intraflagellar transport complex that is conserved in motile flagella and sensory cilia.

Intraflagellar transport (IFT) is an evolutionarily conserved mechanism thought to be required for the assembly and maintenance of all eukaryotic cilia and flagella. Although IFT proteins are present in cells with sensory cilia, the organization of IFT protein complexes in those cells has not been analyzed. To determine whether the IFT complex is conserved in the sensory cilia of photo-receptors, we investigated protein interactions among four mammalian IFT proteins: IFT88/Polaris, IFT57/Hippi, IFT52/NGD5, and IFT20. We demonstrate that IFT proteins extracted from bovine photoreceptor outer segments, a modified sensory cilium, co-fractionate at approximately 17 S, similar to IFT proteins extracted from mouse testis. Using antibodies to IFT88 and IFT57, we demonstrate that all four IFT proteins co-immunoprecipitate from lysates of mouse testis, kidney, and retina. We also extended our analysis to interactions outside of the IFT complex and demonstrate an ATP-regulated co-immunoprecipitation of heterotrimeric kinesin II with the IFT complex. The internal architecture of the IFT complex was investigated using the yeast two-hybrid system. IFT20 exhibited a strong interaction with IFT57/Hippi and the kinesin II subunit, KIF3B. Our data indicate that all four mammalian IFT proteins are part of a highly conserved complex in multiple ciliated cell types. Furthermore, IFT20 appears to bridge kinesin II with the IFT complex.

Adenosine Triphosphate↗
PubMed1988

Localization of a highly divergent mammalian testicular alpha tubulin that is not detectable in brain.

Sequence analysis of a mouse testicular alpha-tubulin partial cDNA, pRD alpha TT1, reveals an isotype that differs from both the somatic and the predominant testicular alpha tubulins at approximately 30% of the 212 amino acid residues determined. Although this mouse testicular cDNA retains the highly conserved sequence, Glu-Gly-Glu-Glu, found in the carboxyl termini of many alpha tubulins, the protein extends substantially beyond this sequence and does not terminate with a C-terminal tyrosine. Using rabbit antiserum prepared to a novel synthetic peptide predicted from this mouse testis alpha-tubulin cDNA, we have have detected by immunoblot and indirect immunofluorescence an antigenic epitope present in testicular alpha tubulin that is not detectable in brain alpha tubulins. We find that the antiserum specifically binds to the manchettes and meiotic spindles of the mouse testis but not with neural fibers or tubulin extracts of the adult mouse brain. These results demonstrate that at least one of the multiple alpha-tubulin isotypes of the mammalian testis is expressed and used in male germ cells but not in the brain.

Amino Acid Sequence↗
PubMed1997

Stage-specific expression of a mouse homologue of the porcine 135kDa alpha-D-mannosidase (MAN2B2) in type A spermatogonia.

The cDNA encoding a mouse homologue of porcine epididymis-specific 135kDa alpha-D-mannosidase (MAN2B2, D28521) was cloned from the mouse testis cDNA library. It was found that 1018 amino acids were coded in its open reading frame, and 62% of the amino acid sequence was identical to that of porcine MAN2B2. In the adult mouse, testis contained higher amounts of mRNA encoding the MAN2B2 homologue than the epididymis, though porcine MAN2B2 was mainly expressed in the narrow region between the caput and corpus epididymis. mRNA of the mouse MAN2B2 homologue was localized exclusively in spermatogonia in the testis. It was specifically expressed in type A spermatogonia at stages IX-XI of spermatogenesis and was detected there until the cell developed into type B spermatogonia. We conclude that the expression of the MAN2B2 homologue can serve as a good marker for the late stages of type A spermatogonia and may have an important role to play in the early step of spermatogenesis in mice.

Amino Acid Sequence↗
PubMed1996

T-type Ca2+ channels and alpha1E expression in spermatogenic cells, and their possible relevance to the sperm acrosome reaction.

There is pharmacological evidence that Ca2+ channels play an essential role in triggering the mammalian sperm acrosome reaction, an exocytotic process required for sperm to fertilize the egg. Spermatozoa are small terminally differentiated cells that are difficult to study by conventional electrophysiological techniques. To identify the members of the voltage-dependent Ca2+ channel family possibly present in sperm, we have looked for the expression of the alpha 1A, alpha 1B, alpha 1C, alpha 1D and alpha 1E genes in mouse testis and in purified spermatogenic cell populations with RT-PCR. Our results indicate that all 5 genes are expressed in mouse testis, and in contrast only alpha 1E, and to a minor extent alpha 1A, are expressed in spermatogenic cells. In agreement with these findings, only T-type Ca2+ channels sensitive to the dihydropyridine nifedipine were observed in patch-clamp recordings of pachytene spermatocytes. These results suggest that low-threshold Ca2+ channels are the dihydropyridine-sensitive channels involved in the sperm acrosome reaction.

Acrosome↗
PubMed1981

Testicular DNA synthesis in vivo: comparison between unilaterally cryptorchid testis and contralateral intact testis in mouse.

DNA synthetic activity was studied by autoradiography in unilaterally cryptorchid (abdominal) and contralateral intact(scrotal) testes in mice. Nongerm cells were inactive in DNA synthesis in both the cryptorchid testis and in the intact control. Labeling indices of undifferentiated type A spermatogonia did not show any differences, indicating that type A spermatogonia did not temperature sensitive in DNA synthesis in vivo.

Animals↗
PubMed2003

Very long chain PUFA in murine testicular triglycerides and cholesterol esters.

Very long chain (VLC) PUFA of the n-6 and n-3 series are known to occur in mammalian testis. The aim of this work was to characterize further two testicular lipid classes with VLCPUFA, cholesterol esters (CE) and total triglycerides (TG) in rat and mouse testis. The VLCPUFA predominating in these lipids were a series of n-6 pentaenes and tetraenes with 24 to 32 carbons, including small amounts of odd-chain PUFA, 28:5n-6 and 24:5n-6 prevailing in CE and TG, respectively. Most of the VLCPUFA of TG were concentrated in a small fraction of TG, made up by 1-O-alkyl-2,3-DAG. This TG subclass was absent altogether from the TG of sexually immature testis. The TG and the CE with VLCPUFA only occurred in testis of adult fertile animals. The proportion of VLCPUFA in total TG and CE was higher in rodents than in other mammals. In the n-6 PUFA-rich adult mouse testis, the amounts of testicular triacylglycerols decreased significantly after consumption of fish oil for 2 wk. Whereas 18:2n-6 was significantly reduced, the amounts of 22:5n-6 and longer n-6 PUFA were less affected in all major testicular lipids including PC and PE, where they were unchanged. The 1-O-alkyl-2,3-DAG and their n-6 VLCPUFA were virtually unaffected by the diet. The VLCPUFA-containing molecular species of CE and TG may represent a form of storage of cholesterol and polyenoic FA required to sustain spermatogenesis. Via chain-shortening, VLCPUFA stored in the neutral lipids may serve as precursors of the major C22 PUFA typical of cell membrane glycerophospholipids, protecting testicular cells against shifts in FA composition induced by dietary changes.

Animals↗
PubMed2004

KIF2Abeta: A kinesin family member enriched in mouse male germ cells, interacts with translin associated factor-X (TRAX).

Translin associated factor X (TRAX) is a binding partner of TB-RBP/Translin. A cDNA encoding the 260 C-terminal amino acids of KIF2Abeta was isolated from mouse testis cDNAs in a yeast two-hybrid library screen for specific TRAX-interacting proteins. KIF2Abeta was expressed predominantly in the mouse testis and enriched in germ cells. The interaction of full-length KIF2Abeta or its C-terminus with TRAX was verified using in vitro synthesized fusion proteins. Deletion mapping of the TRAX-binding region of KIF2Abeta indicated that amino acids 514-659 were necessary and sufficient for the interaction in vivo. Confocal microscopy studies using GFP-fusion proteins demonstrated that KIF2Abeta colocalizes with TRAX in a perinuclear location. KIF2Abeta does not interact with TB-RBP, suggesting that either TRAX can function as an adaptor molecule for motor proteins and TB-RBP, or that this interaction reveals an undescribed role for TRAX in germ cells. The interaction with KIF2Abeta suggests a role for TRAX in microtubule-based functions during spermatogenesis.

Animals↗
PubMed2000

The spatial and temporal expression of Tekt1, a mouse tektin C homologue, during spermatogenesis suggest that it is involved in the development of the sperm tail basal body and axoneme.

Tektins comprise a family of filament-forming proteins that are known to be coassembled with tubulins to form ciliary and flagellar microtubules. Recently we described the sequence of the first mammalian tektin protein, Tekt1 (from mouse testis), which is most homologous with sea urchin tektin C. We have now investigated the temporal and spatial expression of Tekt1 during mouse male germ cell development. By in situ hybridization analysis TEKT1 RNA expression is detected in spermatocytes and in round spermatids in the mouse testis. Immunofluorescence microscopy analysis with anti-Tekt1 antibodies showed no distinct labeling of any subcellular structure in spermatocytes, whereas in round spermatids anti-Tekt1 antibodies co-localize with anti-ANA antibodies to the centrosome. At a later stage, elongating spermatids display a larger area of anti-Tektl staining at their caudal ends; as spermiogenesis proceeds, the anti-Tekt1 staining disappears. Together with other evidence, these results provide the first intraspecies evidence that Tekt1 is transiently associated with the centrosome, and indicates that Tekt1 is one of several tektins to participate in the nucleation of the flagellar axoneme of mature spermatozoa, perhaps being required to assemble the basal body.

Animals↗
PubMed1996

Spermatocyte-specific expression of the gene for mouse testis-specific transcription elongation factor S-II.

Previously, we characterized a rat cDNA for testis-specific transcription elongation factor S-II (SII-T1) (Q. Xu et al., J. Biol. Chem. 269, 3100-3103 (1994)). Here, we isolated a 335-bp fragment of the cDNA for mouse SII-T1, and used it to examine the expression of the SII-T1 gene in the testis by in situ hybridization. The results indicated that the SII-T1 gene is expressed exclusively in spermatocytes, showing no appreciable expression in spermatogonia, spermatids, or Leydig cells. RT-PCR experiments using testis RNA from W/Wv mutant mice also suggested that SII-T1 is a specific transcription elongation factor essential for spermatogenesis.

Amino Acid Sequence↗
PubMed1999

Tlk, a novel evolutionarily conserved murine serine threonine kinase, encodes multiple testis transcripts.

Hypothesizing that genes important in meiotic processes in mammals might have evolutionarily conserved counterparts in lower organisms, we used the yeast IME2 meiotic gene (serine threonine kinase) as a probe for screening a mouse testis cDNA library. This screening resulted in identification of a novel putative serine threonine kinase. Although it did not exhibit significant homology to IME2, it did show significant sequence homology to the Tousled kinase in Arabidopsis. Tousled is associated with various differentiative processes including differentiation of the reproductive organs. The new murine gene was designated accordingly Tlk (Tousled like kinase). Tousled like kinase sequences have been reported to occur in C. elegans and in the human. Positive hybridization signals obtained in zooblot analysis suggest evolutionary conservation of Tlk throughout the phylogenetic ladder. Four distinct Tlk transcripts were detected in mouse testis, at least one of which is testis-specific. Northern and in situ hybridization analyses revealed that in normal testis, Tlk is expressed predominantly in pachytene spermatocytes and in round spermatids. Transcripts differ from one another in their 3' untranslated region, resulting from use of different polyadenylation sites, and in the length of their 5' region. Within the coding region, three of the putative peptides share the kinase and C-terminal domains but differ in their N-terminal domain, suggesting that the latter may be involved in the regulation of Tlk's function. We conclude that although Tlk might have an essential role in all tissues, these kinases are likely to take part in the complex array of phosphorylations involved in regulating spermatogenesis.

Aging↗