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At least 271 records · Page 15Linked to original sources
PubMed2000

Developmental and hormonal regulation of the expression of oligodendrocyte-specific protein/claudin 11 in mouse testis.

The proliferation and differentiation of testicular progenitor stem cells into highly specialized germ cells (spermatozoa) are largely controlled by the hormonally (FSH and testosterone) regulated adjacent supporting Sertoli cells. However, the factors involved in this control remain largely unknown. In the present study, the technique of differential display PCR was used to identify target transcripts to FSH action in cultured murine Sertoli cells. Among these target transcripts, we identified the oligodendrocyte-specific protein (OSP), also known as claudin 11, which had recently been shown to play a key role in the formation of the hematotesticular barrier. Our data show that the testicular expression of OSP is dependent upon male gonad development and systemic and local signaling molecules. Indeed, OSP is expressed early in fetal development in Sertoli cells, immediately after the peak of SRY (sex-determining region, Y gene) expression, but just before that of the anti-Mullerian hormone. Postnatally, OSP expression starts to increase from day 3 to reach a plateau between days 6 and 16 postnatally. In the prepubertal and adult testes, an apparent decline in OSP messenger RNA (mRNA) levels was found, probably because of the increasing number of germ cells (which do not express OSP). Among the signaling molecules that control testicular OSP expression, we have identified FSH and tumor necrosis factor-alpha (TNFalpha). Indeed, using a model of purified cultured mouse Sertoli cells, we demonstrate that FSH inhibits, in a dose (ED50 = 4 ng/ml)- and time (maximal effect after 24 h)-dependent manner, the levels of OSP mRNA. Such an inhibitory effect was mimicked by 8-bromo-cAMP, suggesting that FSH may use the cAMP/protein kinase A pathway to inhibit OSP mRNA levels. TNFalpha was also shown to inhibit OSP expression in cultured Sertoli cells. The maximal effect was observed after 48 h of TNFalpha treatment with an ED50 of 4.5 ng/ml. Together, our results indicate that OSP expression 1) starts during fetal life at a critical period, probably under SRY control and during testicular formation; and 2) is regulated by hormones (FSH) and cytokines (TNFalpha) in the adult testis, suggesting a critical role for these molecules in the (re)modeling process of the hematotesticular barrier during spermatogenesis.

Animals↗
PubMed1999

Morphological alterations in mouse testis by a single dose of malathion.

Malathion((R)) is a widely used organophosphorate agropesticide. In spite of its low toxicity for mammalian cells, it provokes cytogenetic and genotoxic damage both in vivo and in vitro. The effect of Malathion was analyzed in CF-1 young adult male mice. Commercial Malathion (96.6% purity) was injected intraperitoneally in a single dose (250 mg/kg body weight corresponding to 1/12 LD50). Four, 14, 18, and 26 days after injection animals were sacrificed to study epididymal sperm (count and morphology), testicular histology (percentage of depleted seminiferous tubules), and ultrastructural alterations in the germinal epithelium. The effect of Malathion on different germinal cell populations was studied. Teratozoospermia was induced by Malathion at all times studied. Spermatozoa midpiece and flagella were the most affected and at day 18 we observed less alterations of the head. The sperm count at different time intervals was significatively increased compared to controls and there was a parallel increase in depletion of the seminiferous tubules. In conclusion, all germinal cell populations studied were affected by Malathion. Malathion has a teratogenic effect on mice spermatid differentiation, which compromises mostly the flagella, perhaps due to an alkylating effect that disturbs the normal assembling of tail structural protein components. Apparently, the pachytene spermatocyte stage may be relatively more resistant to the pesticide. The Sertoli cells were affected by the insecticide and their damage at an ultrastructural level is highly significant. Cytoplasmatic vacuolization probably revealed metabolic alteration of these cells.

Animals↗
PubMed1998

The identification and characterization of expression of Pftaire-1, a novel Cdk family member, suggest its function in the mouse testis and nervous system.

We have isolated a murine cDNA encoding for a novel putative Cdk-related protein kinase, which has been named Pftaire-1, by screening a testis cDNA library for new serine/threonine kinases. Pftaire-1 showed 50% and 49% amino acid identity with Cdk5 and Pctaire-3, respectively, and contains the eleven subdomains characteristic of the protein kinases. By northern blot analysis we detected two transcripts of approximately 5.5 and 4.9 kb in size. These transcripts were expressed at low level in all murine tissues tested, except in the brain, testis and embryo, where high expression was detected. Cellular localization of the mRNAs by in situ hybridization analysis shows that Pftaire-1 is expressed in late pachytene spermatocytes in the testis and in post mitotic neuronal cells both in the brain and the embryo, suggesting a role of Pftaire-1 both in the process of meiosis as well as neuron differentiation and/or function.

Amino Acid Sequence↗
PubMed2005

Annexin XI co-localises with calcyclin in proliferating cells of the embryonic mouse testis.

Mammalian sex determination relies on the expression of SRY, which triggers a tightly regulated cascade of gene expression leading to male differentiation. Many elements of this pathway remain to be identified. Here, we characterise Annexin XI (Anxa11), a gene whose major site of embryonic expression was within the undifferentiated and differentiating testis. Lower level expression was also observed in both sexes in the Müllerian and Wolffian ducts, the somitic dermamyotome, and the dorsal intermediate zone of the neural tube. Anxa11 transcripts were detected in the indifferent gonad from 10.5 days post coitum (dpc), becoming male specific as development proceeded. Expression was within the testis cords, initially in germ cells, and then in both Sertoli and germ cells. Annexin XI protein was seen in the testis cords from 12.5 dpc, localising to the cytoplasm of the Sertoli cells. Expression of calcyclin (S100a6), shown previously to interact with annexin XI in vitro, was also observed in proliferating cells of the embryonic testis, supporting a possible in vivo interaction.

Animals↗
PubMed1984

Lack of effect of dibromochloropropane on the mouse testis.

A single intraperitoneal injection of (101 X C3Hf)F male mice with 110 mg/kg of 1,2-dibromo-3-chloropropane (DBCP) showed no effect on spermatogonial stem cells or on differentiating spermatogonia as measured by cell counts made 3, 5, and 8 days after injection.

Animals↗
PubMed1993

Sites of transcription of the Müllerian inhibiting substance gene in mouse testis.

We have examined the transcription of Müllerian inhibiting substance (MIS) in testis by the sensitive technique of reverse transcription polymerase chain reaction (RTPCR). A developmental study of testis by this nonquantitative technique showed expression at all postnatal stages, including adults while liver and kidney provided negative controls. Cell separation studies indicated that highly purified interstitial cells, as well as less homogeneous Sertoli cell-enriched and germ cell-enriched fractions, contained RNA for MIS. The transcription of MIS in an interstitial cell type was confirmed by finding MIS mRNA in purified Leydig cells. Inasmuch as the germ cell-enriched fraction contains some Sertoli cells, and XX,Sxra and XX,Sxrb which have germ cell-depleted testes, contain MIS mRNA, a Sertoli cell source remains likely for the seminiferous tubule compartment.

Animals↗
PubMed2002

Direct injection of foreign DNA into mouse testis as a possible in vivo gene transfer system via epididymal spermatozoa.

We have attempted to transfect testicular spermatozoa with plasmid DNA by direct injection into testes to obtain transgenic animals [this technique was thus termed "testis-mediated gene transfer (TMGT)"]. When injected males were mated with superovulated females 2 and 3 days after injection, (i) high efficiencies (more than 50%) of gene transmission were achieved in the mid-gestational F0 fetuses, (ii) the copy number of plasmid DNA in the fetuses was estimated to be less than 1 copy per diploid cell, and (iii) overt gene expression was not found in these fetuses. These findings suggest the possibility that plasmid DNA introduced into a testis is rapidly transported to the epididymis and then incorporated by epididymal spermatozoa. The purpose of this study was to elucidate the mechanism of TMGT by introducing trypan blue (TB) or Hoechst 33342 directly into testis. We found that TB is transported to the ducts of the caput epididymis via rete testis within 1 min after testis injection, and TB reached the corpus and cauda epididymis within 2-4 days after injection. Staining of spermatozoa isolated from any portion of epididymis was observed 4 days after injection of a solution containing Hoechst 33342. Injection of enhanced green fluorescent protein (EGFP) expression vector/liposome complex into testis resulted in transfection of epithelial cells of epididymal ducts facing the lumen, although the transfection efficiency appeared to be low. In vivo electroporation toward the caput epididymis immediately after injection of EGFP expression vector into a testis greatly improved the uptake of foreign DNA by the epididymal epithelial cells. PCR analysis using spermatozoa isolated from corpus and cauda epididymis 4 days after injection of a DNA/liposome complex into testis revealed exogenous DNA in these spermatozoa even after treatment with DNase I. These findings indicate that exogenous DNA introduced into tesits is rapidly transported to epididymal ducts via the rete testis and efferent ducts, and then incorporated by epithelial cells of epididymis and epididymal spermatozoa.

Animals↗
PubMed1997

p53 is associated with the nuclear envelope in mouse testis.

p53 has been postulated to play a role in meiosis as well as in the regulation of germ cell numbers by apoptosis. This study investigated the subcellular localization of p53 in the testis, including conditions known to induce germ cell apoptosis. Western blot analysis showed that p53 was enriched in the nuclear envelope fraction, and confocal microscopy confirmed that p53 was associated with the nuclear envelope of germ cells. Exposure of the testis to heat stress induced translocation of p53 into the nucleus. Nuclear envelope binding provides an optimal site for rapid entry of p53 into the nucleus, where it may act as a DNA-binding protein to induce apoptosis or cell cycle arrest in response to appropriate stimuli. The nuclear envelope sequestration of p53 also provides a framework to understand how mitosis and meiosis in the testis may proceed despite high intracellular concentration of p53.

Animals↗
PubMed1984

Germ cell survival, differentiation, and epididymal transit kinetics in mouse testis subjected to high in vivo levels of testosterone enanthate.

Androgenic steroids induce oligospermia at high doses, but few studies have assessed which cell types and spermatogenic processes are affected. Testis cell numbers and rate of differentiation were quantitatively evaluated in testosterone enanthate-treated male mice by use of histological and cell separation techniques. Reduction in the number of testicular germ cells in treated animals accounted for 40% of the total reduction of spermatozoa. Testis cells differentiated at a similar rate in treated and control animals. Spermatozoa moved to the cauda epididymidis a day earlier in testosterone-treated animals, and were lost from the cauda after only two days, rather than after the normal seven days. These results suggest that over half of the 98% reduction in sperm number in testosterone-treated animals occurred at the stage of passage through the cauda epididymidis.

Animals↗