Quantification of apoptosis (programmed cell death) in mammalian testis by DNA-fragmentation ELISA

Hingst, O.; Blöttner, S.

doi:10.1016/0093-691x(95)00186-c

Cited by 26 publications

(17 citation statements)

References 19 publications

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“…An ELISA method has also been adapted by which monoclonal antibodies are directed against the histones and the DNA fragments resulting from apoptosis [13]. Recently, flow cytometry has been used to detect apoptosis in granulosa cells from porcine follicles [9].…”

Section: Introductionmentioning

confidence: 99%

Analysis of Atresia in Bovine Follicles Using Different Methods: Flow Cytometry, Enzyme-Linked Immunosorbent Assay, and Classic Histology1

Blondin

Dufour

Sirard

1996

Biology of Reproduction

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This study was performed in order to validate flow cytometry as an acceptable method for analyzing follicular atresia in bovine granulosa cells by comparing it to two other techniques, histology and ELISA. Ovaries from 35 nontreated cows, all at different times of their estrous cycle, and 12 superovulated cows were collected. Superovulation treatments began between Days 9 and 12 (Day 0 = estrous), and animals were administered 8 doses of FSH-P (32 or 20 mg) at 12-h intervals over 4 days with or without the addition of 1 mg of prostaglandin s.c. on the third day. Animals were slaughtered after the last FSH-P injection. Granulosa cells from 133 follicles from non-treated cows and 85 follicles from superovulated cows were analyzed. Follicular diameters ranged from 2 to 20 and 2 to 16 mm, respectively. Because of the ample amounts of cells collected, it was possible to perform more than one technique for each follicle. Flow cytometry detected in most follicles a subpopulation of cells that possessed less DNA than normal, viable cells (referred to as -G1 cells). Histological classes used (established in previous work) were nonatretic (< or = 5% picnotic nuclei), slightly atretic (> 5 to < 15% picnotic nuclei), and atretic (> or = 15% picnotic nuclei). A strong linear correlation existed between the percentage of picnotic nuclei and the percentage of -G1 cells (R = 0.86; p < 0.001) with granulosa cells from follicles from nontreated cows. In some cases, flow cytometry detected a certain percentage of cells with reduced DNA content while histology revealed very few picnotic nuclei, indicating a higher sensitivity of flow cytometry. Superovulation decreased considerably the percentage of atretic cells seen with both techniques. The linear correlation was not as strong because follicles from superovulated animals represent a very homogenous population (R = 0.54; p < 0.001). The ELISA technique coincided with flow cytometry as seen in the strong correlation between the two techniques (R = 0.91; p < 0.001). Flow cytometry appeared to be very effective and rapid in evaluating the atretic states of follicles from nontreated and superovulated cows. Strong correlations existed between this method and histology and ELISA.

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Section: Introductionmentioning

confidence: 99%

Analysis of Atresia in Bovine Follicles Using Different Methods: Flow Cytometry, Enzyme-Linked Immunosorbent Assay, and Classic Histology1

Blondin

Dufour

Sirard

1996

Biology of Reproduction

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“…We used two independent investigation methods: in situ, using TUNEL assay and a quantitative ELISA assay, based on the detection of free nucleosomes in retinal extracts [32]. An increase in the number of TUNEL-positive cells was observed in the ONL with only a few TUNEL-positive cells being observed in the inner retina after light exposure (Fig.…”

Section: Resultsmentioning

confidence: 99%

Thioredoxin rod-derived cone viability factor protects against photooxidative retinal damage

Elachouri

Lee-Rivera

Clérin

et al. 2015

Free Radical Biology and Medicine

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Rod-derived cone viability factor (RdCVF) is a trophic factor of the thioredoxins family that promotes the survival of cone photoreceptors. It is encoded by the nucleoredoxin-like gene 1 Nxnl1 which also encodes by alternative splicing a long form of RdCVF (RdCVFL), a thioredoxin enzyme that interacts with TAU. The known role of thioredoxins in the defense mechanism against oxidative damage led us to examine the retinal phenotype of the Nxnl1(-/-) mice exposed to photooxidative stress. Here we found that, in contrast to wild-type mice, the rod photoreceptors of Nxnl1(-/-) mice are more sensitive to light after exposure to 1700 or 2500 lx. The delivery of RdCVF by AAV to mice deficient of Nxnl1(-/-) protects rod photoreceptors from light damage. Interestingly, the RdCVF2L protein, encoded by the paralog gene Nxnl2, is able to reduce TAU phosphorylation, as does RdCVFL, but does not protect the rod from light damage. Our result shows that the Nxnl1 gene, through the thioredoxin RdCVFL, is part of an endogenous defense mechanism against photooxidative stress that is likely of great importance for human vision.

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“…The same authors also showed that apoptotic germ cells are phagocytized by Sertoli cells [Seco-Rovira et al, 2015]. An inverse rela- tionship between germ cell proliferation and apoptosis has been found during seasonal testicular changes in roe deer [Blottner et al, 1995[Blottner et al, , 1996Schön et al, 2004]. Nevertheless, in this species it was later found that apoptosis does not cause massive germ cell depletion taking place during testis regression [Blottner et al, 2007].…”

Section: Cellular Mechanisms Of Testis Regression In Vertebrates: Thementioning

confidence: 97%

“…3 ). Several authors reported that apoptosis contributes to testicular regression in the European brown hare [Strbenc et al, 2003;Blottner et al, 1995] and hamsters [Furuta et al, 1994;Morales et al, 2002Morales et al, , 2007. However, conclusive data in the Syrian hamster was only recently provided by Seco-Rovira et al [2014] showing that apoptotic incidence peaks during the period of testis regression and not later when the process has been completed, and the increase is high enough to explain a massive depletion of the germinative epithelium.…”

Section: Cellular Mechanisms Of Testis Regression In Vertebrates: Thementioning

confidence: 97%

“…Available data indicate that apoptosis could plausibly mediate testis regression in birds Jenkins et al, 2007;Islam et al, 2012], turtles [Zhang et al, 2008], most amphibians [Yazawa et al, 1999[Yazawa et al, , 2000, and some mammals [Seco-Rovira et al, 2014. Contrarily, in one frog species [Sasso-Cerri et al, 2006] and several mammals [Young et al, 1999;Hingst and Blottner, 1995;Nonclercq et al, 1996] apoptosis was associated with but not conclusively shown to trigger testis regression [Dadhich et al, 2013], and in others apoptosis is not the effector of testis regression [Blottner et al, 2007;Dadhich et al, 2010;Luaces et al, 2014]. In conclusion, apoptosis is not a general mechanism of seasonal testis regression mainly in mammals, which is not a homogeneous group in this respect.…”

Section: Cellular Mechanisms Of Testis Regression In Vertebrates: Thementioning

confidence: 99%

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Circannual Testis Changes in Seasonally Breeding Mammals

Jiménez

Burgos

Barrionuevo³

2015

Sex Dev

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In the non-equatorial zones of the Earth, species concentrate their reproductive effort in the more favorable season. A consequence of seasonal breeding is seasonal testis regression, which implies the depletion of the germinative epithelium, permeation of the blood-testis barrier, and reduced androgenic function. This process has been studied in a number of vertebrates, but the mechanisms controlling it are not yet well understood. Apoptosis was assumed for years to be an important effector of seasonal germ cell depletion in all vertebrates, including mammals, but an alternative mechanism has recently been reported in the Iberian mole as well as in the large hairy armadillo. It is based on the desquamation of meiotic and post-meiotic germ cells as a consequence of altered Sertoli-germ cell adhesion molecule expression and distribution. Desquamated cells are either discarded alive through the epididymis, as in the mole, or subsequently die by apoptosis, as in the armadillo. Also, recent findings on the reproductive cycle of the greater white-toothed shrew at the meridional limits of its distribution area have revealed that the mechanisms controlling seasonal breeding are in fact far more plastic and versatile than initially suspected. Perhaps these higher adaptive capacities place mammals in a better position to face the ongoing climate change.

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Quantification of apoptosis (programmed cell death) in mammalian testis by DNA-fragmentation ELISA

Cited by 26 publications

References 19 publications

Analysis of Atresia in Bovine Follicles Using Different Methods: Flow Cytometry, Enzyme-Linked Immunosorbent Assay, and Classic Histology1

Analysis of Atresia in Bovine Follicles Using Different Methods: Flow Cytometry, Enzyme-Linked Immunosorbent Assay, and Classic Histology1

Thioredoxin rod-derived cone viability factor protects against photooxidative retinal damage

Circannual Testis Changes in Seasonally Breeding Mammals

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