Testicular apoptosis in feral Clarias gariepinus using TUNEL and cleaved caspase-3 immunohistochemistry

McClusky, Leon Mendel; Barnhoorn, I.E.J.; Dyk, J C van; Bornman, M. S.

doi:10.1016/j.ecoenv.2008.03.017

Cited by 20 publications

(15 citation statements)

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“…Thereafter, the nucleus or the chromatin masses lose caspase-3 immunoreactivity and stain intensely blue-purple, a color similar to that of the condensed chromatin in spermatid nuclei, and is which proof that the chromatin in the apoptotic germ cell is indeed now condensed. All of these stages of the death process are TUNEL-negative in all three species, as has been previously reported (McClusky et al 2007(McClusky et al , 2008. By contrast, subsequent phases of the death process, i.e.…”

Section: Discussionmentioning

confidence: 57%

“…For comparative analysis, testes were obtained from four men, aged 57-70 years old, who underwent orchidectomy as treatment for advanced prostate cancer but otherwise healthy with no known endocrine disease, as described in McClusky et al (2007), and ten laboratory-raised sexually mature adult feral sharptooth catfish (Clarias gariepinus), sampled and processed as described in McClusky et al (2008). These human testicular tissues were specifically used because they were the only readily available material.…”

Section: Organisms and Tissue Samplingmentioning

confidence: 99%

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Immunohistochemical study of nuclear changes associated with male germ cell death and spermiogenesis

et al. 2009

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In a previous study on the effects of gestational and lactational exposure of para-nonylphenol on male rats, we noted in both induced and uninduced rats, that variations in cleaved caspase-3 immunostaining patterns were associated with distinct nuclear alterations in mainly basally located germ cells (spermatogonia and preleptotene spermatocytes). These were re-analysed and compared with cleaved caspase-3-labeled germ cells in the aging human and the spermatogenically active catfish testis. In the rat testes, cytoplasmic immunostaining was progressively associated with lateral compression of the nucleus, its break up into large pieces which can contain immunostained marginated chromatin masses. The pale remnants of the nucleus continued to shrink in size concomitant with the appearance of blue-purplish stained regions in the cytoplasm similar in color to the condensed chromatin in spermatids, a condition which was TUNEL-negative. These large clumps of chromatin also eventually disappeared, giving rise to cells resembling cytoplasmic ghosts, a condition which was TUNEL-positive. By contrast, the immunolabeled nuclei of human and catfish germ cells condensed into a single mass, after which they lost immunoreactivity. To exclude the possibility that these observations could reflect alterations in Sertoli nuclei, rat testicular sections were probed with a mouse anti-human GATA-4 monoclonal (MHM) antibody. The MHM was, however, the second of two GATA-4 antibodies tested, with a goat anti-mouse polyclonal (GMP) initially used to label the rat Sertoli nuclei. GMP unexpectedly, but distinctly labeled the complete development of the acrosome in the rat testis, a fortuitous finding with utility for staging of the seminiferous epithelium.

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Section: Discussionmentioning

confidence: 57%

Section: Organisms and Tissue Samplingmentioning

confidence: 99%

Immunohistochemical study of nuclear changes associated with male germ cell death and spermiogenesis

et al. 2009

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“…This condition has, in rodents at least, often been ascribed to the rapid and efficient phagocytic activity of the closely positioned Sertoli cells (Russell & Clermont 1977, Bartke 1995, Billig et al 1995. All of this is in stark contrast to that seen in lower vertebrates where entire germinal clones enter apoptosis (Yazawa et al 2003, Corriero et al 2007, McClusky et al 2008). …”

Section: Introductionmentioning

confidence: 99%

“…Procedures previously optimised for fish testicular tissues were followed (McClusky et al 2008). Briefly paraffin sections (4-5 mm thick) were cut and collected onto superfrost plus glass slides, deparaffinised and rehydrated stepwise through an ethanol series.…”

Section: Cleaved Caspase-3 Immunohistochemistrymentioning

confidence: 99%

“…Negative controls were generated by omitting the primary antibody and by serial dilution of the primary antibody. For positive controls, testicular sections from the thresher shark (A. vulpinus) caught at the same time and catfish, C. gariepinus (McClusky et al 2008), were processed simultaneously. Figure 10 Ventral view of the testis of the adult male blue shark.…”

Section: Cleaved Caspase-3 Immunohistochemistrymentioning

confidence: 99%

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The caspase-dependent apoptosis gradient in the testis of the blue shark, Prionace glauca

McClusky¹

2013

Reproduction

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The severe degenerative phenomena that characterises spermatogenesis in mating blue sharks involves spatially separated germ cell and Sertoli cell apoptosis. Unlike that observed in multilayered type B spermatogonial and spermatocyte cysts caspase-3-dependent apoptosis of single and multinucleate type B spermatogonia in one to three spermatogonial layered cysts resulted in their complete fragmentation, delayed phagocytic removal and displacement of the apoptotic bodies towards the perilumenar Sertoli nuclei. Changes were observed in the immunostaining patterns of proliferating cell nuclear antigen (PCNA), including subtle changes in cytoplasmic and overall intense immunostaining, labelled single and multinucleate cell (MNC) apoptotic spermatogonial masses in premeiotic cysts in different stages of the protracted death process. Initial massive MNC formation at the mitosis-meiosis transition eventually left its imprint in the spermatogenic sequence in the form of vacuolated areas in the affected and subsequent stages. Some of the latter attempted further developmental advance but eventually degenerated. The observed higher PCNA index of spermatogonia in vacuolated testes compared to testes with the MNC type of degeneration indicated that the former testicular morphology represented, in essence, the recovery phase from the pronounced MNC death earlier. Events culminating in the eventual apoptotic demise of the Sertoli cells themselves included the abortion of further development (presumably due to a suboptimal Sertoli:germ cell ratio) of those germ cells left over from the wave of MNC death that swept the cysts. Eventually the Sertoli-cell-only cysts became apoptotic as they were engulfed by the infiltrating lymphomyeloid cells from the epigonal organ associated with the mature pole of the testis.

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Role of HSP70 in the regulation of the testicular apoptosis in a seasonal breeding teleost Prochilodus argenteus from the São Francisco river, Brazil

Domingos

Thomé

Martinelli

et al. 2013

Microscopy Res & Technique

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This study investigated the relationship among heat shock protein 70 (HSP70), proliferating cell nuclear antigen (PCNA), and testicular apoptosis during a breeding cycle of Prochilodus argenteus, a neotropical migratory characiform fish of importance in commercial fishery from the São Francisco River basin. A total of 48 (12 fish/sampling) adult males were caught using casting and drifting nets in four samplings from June 2008 to March 2009. Immunohistochemistry, Western blotting, terminal transferase-mediated dUTP nick-end labeling (TUNEL), enzyme-linked immunosorbent assay (ELISA), and caspase-3 colorimetric assay were assessed in different phases of spermatogenesis. Labeling for HSP70 occurred in spermatogonia (SPG(A) 18.0±1.5 and SPGB 27.9±1.0 in 100 mm(2), respectively) and Sertoli cells in all sampling periods, with higher values in June (resting period) while spermatocytes were labeled in September (maturation period) and December (ripe period). For PCNA, immunoreaction was predominant in spermatogonia in June and September, while primary spermatocytes were labeled mainly in December (18.7±2.0). TUNEL-positive reaction occurred throughout the sampling periods, and labeling was detected in the nucleus of germ cells in all developmental phases, except spermatozoa. By ELISA, total HSP70 in testis increased significantly from June to December, and decreased in March (regression period), P<0.05. Caspase-3 activity decreased from June to December and increased in March. Taken together, our results suggest that HSP70 may protect the germ cells from caspase-3-dependent apoptosis during testicular activity and, reduction of HSP70 and increase of apoptosis contribute for testicular remodeling after the breeding season in wild populations of P. argenteus in the São Francisco River.

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Testicular apoptosis in feral Clarias gariepinus using TUNEL and cleaved caspase-3 immunohistochemistry

Cited by 20 publications

References 20 publications

Immunohistochemical study of nuclear changes associated with male germ cell death and spermiogenesis

Immunohistochemical study of nuclear changes associated with male germ cell death and spermiogenesis

The caspase-dependent apoptosis gradient in the testis of the blue shark, Prionace glauca

Role of HSP70 in the regulation of the testicular apoptosis in a seasonal breeding teleost Prochilodus argenteus from the São Francisco river, Brazil

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