Vimentin expression during altered spermatogenesis in rats

Kopecký, Martin; Semecký, Vladimír; Nachtigal, Petr

doi:10.1016/j.acthis.2005.06.007

Cited by 55 publications

(52 citation statements)

References 25 publications

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“…Since vimentin filaments of Sertoli cells play an important role in the maintenance of spermatogenesis 46) , our finding suggests that consecutive exposure of DiBP can induce the conspicuous alterations in distribution of vimentin filaments, and it correlates with sloughing of spermatogenic cells from the seminiferous epithelium. From the present recovery experiment, vimentin filaments can be reorganized after elimination of DiBP, and then spermatogenic cells repopulate and are restored 46) . In conclusion, our present study showed that DiBP caused a significant increase of TUNEL-positive spermatogenic cells and a disorder of vimentin filaments in rats.…”

Section: Discussionmentioning

confidence: 83%

Effects of di-iso-butyl phthalate on testes of prepubertal rats and mice

Zhu

Tay

Andriana

et al. 2010

Okajimas Folia Anat. Jpn.

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Summary: Di-iso-butyl phthalate (DiBP), a special plasticizer, is used as a substitute for di(n-butyl) phthalate(DBP). The effects of DiBP on testes in prepubertal rodents still remain to be obscure. Testicular toxicity of DiBP was investigated in 21-day-old Sprague-Dawley rats and C57BL/6N mice, using with in situ TUNEL method. For an acute exposure experiment, animals were once given DiBP at various concentrations by oral gavage. For a subchronic exposure experiment, they were daily given DiBP at various concentrations for consecutive 7 days. Controls were treated with corn oil under the same condition. For a recovery experiment, rats were once given DiBP (1000 mg/kg), and were sacrificed at day 1 to 8 after administration. Furthermore, the disorder of vimentin filaments in Sertoli cells after daily administration of DiBP (500 mg/kg) for consecutive 7 days in rats also identified by immunohistochemistry using anti-vimentin antibody. As a result, the present study demonstrated that DiBP can induce testicular atrophy in rats due to the increase of TUNEL-positive spermatogenic cells in both acute and subchronic exposure experiments. At the same time, the disorder of vimentin filaments in Sertoli cells was recognized. However, no such damages could be found in mouse testis. For the recovery experiment, the testis weight and testicular morphology returned to normal at day 6 after administration. In conclusion, the present study indicates that DiBP causes the significant increase of TUNEL-positive spermatogenic cells and the disorder of vimentin filaments in Sertoli cells in rats and that DiBP shows a species-specific toxicity.

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Section: Discussionmentioning

confidence: 83%

Effects of di-iso-butyl phthalate on testes of prepubertal rats and mice

Zhu

Tay

Andriana

et al. 2010

Okajimas Folia Anat. Jpn.

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“…Sertoli cells are the major cell type expressing vimentin in the rat testis, but it is also expressed in the interstitial cell compartment in the rat testis (our results, Alam & Kurohmaru, 2014; Kopecky et al ., 2005). More precise dissection of the somatic cell population can be achieved by coupling additional marker antibodies (such as Sox9 for Sertoli cells or 3βHSD for Leydig cells) to the assay.…”

Section: Discussionmentioning

confidence: 99%

A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

et al. 2015

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SummaryAccurate analysis and quantification of different testicular cell populations are of central importance in studies of male reproductive biology. The traditional histomorphometric and immunohistochemical methods remain the gold standard in studying the complex dynamics of the testicular tissue. Through past years advances have been made in the application of flow cytometry for the rapid analysis of testicular cell populations. Detection of DNA content and of surface antigens and fluorescent reporters have been widely used to analyze and sort cells. Detection of intracellular antigens can broaden the possibilities of applying flow cytometry in studies of male reproduction. Here, we report a detailed protocol for the preparation of rat testicular tissue for detection of intracellular antigens by flow cytometry, and a pipeline for subsequent data analysis and troubleshooting. Rat testicular ontogenesis was chosen as the experimental model to validate the performance of the assay using vimentin and γH2AX as intracellular markers for the somatic and spermatogenic cells, respectively. The results show that the assay is reproducible and recapitulates the rat testis ontogenesis.

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“…The number of contaminating germ cells was reduced to a very low level (about 10%) by several cycles of fragmentation of the cell clusters through a 19-gauge syringe needle, followed by sedimentation. Vimentin immunoreactivity was used to determine the percentage of germ cells (vimentin negative) contaminating the (vimentin positive) Sertoli cell preparations (Franke et al 1979, Suter et al 1997, Kopecky et al 2005). An aliquot of each Sertoli cell preparation was cytospun onto aminoalkylsilanized slides.…”

Section: Preparation Of Sertoli and Germinal Cell Fractionsmentioning

confidence: 99%

Glial cell-line-derived neurotropic factor and its receptors are expressed by germinal and somatic cells of the rat testis

Fouchécourt¹,

Godet²,

Sabido³

et al. 2006

Journal of Endocrinology

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Glial cell-line-derived neurotropic factor (GDNF) and its receptors glial cell-line-derived neurotropic factor a (GFR1a) and rearranged during transformation (RET) have been localized in the rat testis during postnatal development. The three mRNAs, and GDNF and GFR1a proteins were detected in testis extracts from 1-to 90-day-old rats by reverse transcriptase PCR and Western blotting respectively. The three mRNAs were present in Sertoli cells from 20-and 55-day-old rats, pachytene spermatocytes (PS), and round spermatids (RS). The GDNF and GFR1a proteins were detected in PS, RS, and Sertoli cells. GDNF and GFR1a were also detected using flow cytometry in spermatogonia and preleptotene spermatocytes, and in secondary spermatocytes. The localization of GDNF and GFR1a in germ and Sertoli cells was confirmed by immunocytochemistry. The hypothesis that GDNF may control DNA synthesis of Sertoli cells and/or spermatogonia in the immature rat was addressed using cultures of seminiferous tubules from 7-to 8-day-old rats. Addition of GDNF for 48 h resulted in a twofold decrease in the percentage of spermatogonia able to duplicate DNA, whereas Sertoli cells were not affected. These results are consistent with a role of GDNF in inhibiting the S-phase entrance of a large subset of differentiated type A spermatogonia, together with an enhancing effect of the factor on a small population of undifferentiated (stem cells) spermatogonia. Moreover, the wide temporal and spatial expression of GDNF and its receptors in the rat testis suggest that it might act at several stages of spermatogenesis.

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Vimentin expression during altered spermatogenesis in rats

Cited by 55 publications

References 25 publications

Effects of di-iso-butyl phthalate on testes of prepubertal rats and mice

Effects of di-iso-butyl phthalate on testes of prepubertal rats and mice

A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

Glial cell-line-derived neurotropic factor and its receptors are expressed by germinal and somatic cells of the rat testis

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