Involvement of Calpain for Apoptosis in Dysfunction of the Unaffected Testis in Rats with Experimental Testicular Torsion

Umemoto, Yukihiro; Sasaki, Shoichi; Tatsura, Hiroyuki; Kubota, Hirokazu; Kubota, Yasue; Kohri, Kenjiro; Ozaki, Yasuhiko; Sasaki, Makoto

doi:10.1111/j.8755-8920.2001.450407.x

Cited by 11 publications

(8 citation statements)

References 27 publications

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“…Based on the observation that testicular damage was found in animals undergoing spermatic cord torsion but not after spermatic vessel ligation, Cerasaro et al [8] suggested that a minimum of blood circulation is necessary to trigger a systemic reaction being able to induce damage in the contralateral testis. Although initial studies reported damage to the contralateral testis in experimental models of spermatic cord torsion [4,7], only recently was the phenomenom of germ cell apoptosis studied [9,10]. Moreover, little attention has been focused on testicular immune cells and their secretion products in this model.…”

Section: Tel: +86-21-5492-2824; Fax: +86-21-5492-2825; Shanghai Chinamentioning

confidence: 99%

Immunohistopathology of the contralateral testis of rats undergoing experimental torsion of the spermatic cord

Rodriguez¹,

Rival²,

Theas³

et al. 2006

Asian J Andrology

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Experimental spermatic cord torsion induces, in the contralateral testis, a focal damage of seminiferous tubules characterized by apoptosis and sloughing of germ cells. Results suggest humoral and cellular immune mediated testicular cell damage in which macrophages and mast cells seem to be involved in the induction of germ cell apoptosis through the TNF-alpha/TNFR1 system and in the modulation of the inflammatory process.

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Section: Tel: +86-21-5492-2824; Fax: +86-21-5492-2825; Shanghai Chinamentioning

confidence: 99%

Immunohistopathology of the contralateral testis of rats undergoing experimental torsion of the spermatic cord

Rodriguez¹,

Rival²,

Theas³

et al. 2006

Asian J Andrology

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“…Calpastatin is an endogenous inhibitor of calpain, and calpain has been shown to be involved in apoptosis. Given the observation that calpain is expressed in the mouse testis and that its staining localizes to rat spermatocyte nuclei after experimental testicular torsion (Umemoto et al, 2001), it seems likely that calpain is normally expressed in mouse spermatocytes or that it is induced to detectable levels in these cells after a stressor, such as heat. It is possible that the suppression of heat‐induced spermatocyte apoptosis by tCAST overexpression is via the inhibition of calpain.…”

Section: Discussionmentioning

confidence: 99%

“…The gene was shown to be transcribed in haploid germ cells and not in spermatocytes, and the protein was shown to associate with intracellular membrane structures of the postmeiotic cells. Thus, whereas calpain is expressed in premeiotic cells (Umemoto et al, 2001), tCAST, under normal circumstances, is not (Li and Goldberg, 2000).…”

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confidence: 99%

Heat‐Induced Apoptosis of Mouse Meiotic Cells Is Suppressed by Ectopic Expression of Testis‐Specific Calpastatin

Somwaru

Li²,

Doglio³

et al. 2004

Journal of Andrology

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Calpastatin is a naturally occurring inhibitor of calpain, a protease involved in apoptotic cell death. A testis-specific isoform of calpastatin (tCAST) has been identified that is transcribed in haploid germ cells but not in spermatocytes. To investigate the possible function(s) of tCAST, we tested the hypothesis that the ectopic expression of calpastatin in spermatocytes would suppress the death of these cells in response to an apoptosis-inducing stimulus in vivo. To this end, the 5'-flanking region of the mouse ldhc gene was linked to tCAST, and transgenic mice were generated. Immunohistochemical analysis revealed that, in contrast to control sections in which the signal for tCAST was seen in round spermatids, intense staining was visualized in pachytene spermatocytes in the transgenic animals, indicating that the strategy we used to generate the transgenic animals resulted in the ectopic expression of tCAST in spermatocytes. We then tested the effect of a short period of heating on germ cell apoptosis in the testes of wild-type and transgenic mice. Pachytene spermatocytes were the major germ cell type seen to undergo apoptosis after heat treatment. There were no differences in the number of apoptotic germ cells per seminiferous tubule between wild-type and tCAST transgenic control mice; thus, there was no apparent effect of the transgene on normal apoptosis. Heating resulted in increased numbers of TUNEL-positive germ cells in both wild-type and tCAST transgenic mice, as well as increased testicular DNA fragmentation. Heating the tCAST transgenic mouse testes resulted in significantly fewer apoptotic cells per seminiferous tubule than in wild-type mice at both 8 and 24 hours after treatment. Thus, as hypothesized, the ectopic expression of tCAST in pachytene spermatocytes suppressed germ cell apoptosis.

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“…For evaluation of apoptosis, the numbers of seminiferous tubules with TUNEL‐positive cells were counted under a light microscope and are expressed here relative to all seminiferous tubules as the apoptosis index (AI) as we described previously (Umemoto et al, 2001). The frequency of apoptotic cells was compared between groups.…”

Section: Methodsmentioning

confidence: 99%

Gene Transfer to Mouse Testes by Electroporation and Its Influence on Spermatogenesis

Umemoto

Sasaki

Kojima

et al. 2005

Journal of Andrology

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We transferred the adventitious gene pCAGGS-lacZ to mouse testes with the use of a square-wave electroporator and investigated the efficiency of gene transfer (GT) and the influence of the procedure on testicular damage and spermatogenesis. Mice were divided into 5 groups: (1-2) injection of gene/phosphate-buffered saline (PBS) into the interstitial space followed by electroporation (EP), (3) EP alone, (4-5) injection of gene/PBS without EP. The presence of the lacZ gene was determined by X-gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) staining and the polymerase chain reaction (PCR). The influence of transfer on spermatogenesis was assessed by evaluating the seminiferous tubules according to the Johnsen score (JS). TdT-mediated dUTP-biotin nick end-labeling (TUNEL) staining was performed for the detection of apoptosis in the testes to evaluate the testicular damage caused by GT, and fertilization ability was assessed by mating male mice from each group with normal female mice at 1, 2, 4, 6, and 8 weeks after the procedure. LacZ expression was detected by X-gal staining and PCR for 4 weeks after GT in group 1. But in group 4, LacZ expression was not detected for all times. In groups 1 through 3, the JSs decreased gradually until 4 weeks and recovered at 6 and 8 weeks after GT. The JSs were significantly decreased at 4 weeks for groups 1 through 3 compared with groups 4 and 5. In groups 1 through 3, apoptotic cells were significantly more numerous at 1, 2, and 4 weeks after the procedure, and there were significant differences in their numbers between groups 1 through 3 and groups 4 and 5 until 4 weeks after the procedure. The number of offspring did not differ significantly between all groups. These results suggest that although spermatogenic damage caused by EP could present problems, GT by EP might be effective for transfecting germ cells or somatic cells and could be applicable for in vivo gene therapy for male infertility in the future.

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Involvement of Calpain for Apoptosis in Dysfunction of the Unaffected Testis in Rats with Experimental Testicular Torsion

Abstract: It was suggested that mu-calpain may be involved in apoptosis expression in sperm formation disorder of the unaffected testis in unilateral testicular torsion.

Cited by 11 publications

References 27 publications

Immunohistopathology of the contralateral testis of rats undergoing experimental torsion of the spermatic cord

Immunohistopathology of the contralateral testis of rats undergoing experimental torsion of the spermatic cord

Heat‐Induced Apoptosis of Mouse Meiotic Cells Is Suppressed by Ectopic Expression of Testis‐Specific Calpastatin

Gene Transfer to Mouse Testes by Electroporation and Its Influence on Spermatogenesis

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