Glutamate Cysteine Ligase Modifier Subunit (Gclm) Null Mice Have Increased Ovarian Oxidative Stress and Accelerated Age-Related Ovarian Failure

Glutathione (GSH) is the one of the most abundant intracellular antioxidants. Mice lacking the modifier subunit of glutamate cysteine ligase (Gclm), the rate-limiting enzyme in GSH synthesis, have decreased GSH. Our prior work showed that GSH plays antiapoptotic roles in ovarian follicles. We hypothesized that Gclm−/− mice have accelerated ovarian aging due to ovarian oxidative stress. We found significantly decreased ovarian GSH concentrations and oxidized GSH/oxidized glutathione redox potential in Gclm−/− vs Gclm+/+ ovaries. Prepubertal Gclm−/− and Gclm+/+ mice had similar numbers of ovarian follicles, and as expected, the total number of ovarian follicles declined with age in both genotypes. However, the rate of decline in follicles was significantly more rapid in Gclm−/− mice, and this was driven by accelerated declines in primordial follicles, which constitute the ovarian reserve. We found significantly increased 4-hydroxynonenal immunostaining (oxidative lipid damage marker) and significantly increased nitrotyrosine immunostaining (oxidative protein damage marker) in prepubertal and adult Gclm−/− ovaries compared with controls. The percentage of small ovarian follicles with increased granulosa cell proliferation was significantly higher in prepubertal and 2-month-old Gclm−/− vs Gclm+/+ ovaries, indicating accelerated recruitment of primordial follicles into the growing pool. The percentages of growing follicles with apoptotic granulosa cells were increased in young adult ovaries. Our results demonstrate increased ovarian oxidative stress and oxidative damage in young Gclm−/− mice, associated with an accelerated decline in ovarian follicles that appears to be mediated by increased recruitment of follicles into the growing pool, followed by apoptosis at later stages of follicular development.

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Follicular development, morphological integrity, and oxidative stress in bovine preantral follicles cultured in vitro with ascorbic acid

Zygote ◽

10.1017/s0967199421000903 ◽

2021 ◽

pp. 1-7

Author(s):

Larissa Zamparone Bergamo ◽

Denis Vinicius Bonato ◽

Camila Bizarro-Silva ◽

Francieli Gesleine Capote Bonato ◽

Tamires Korchovei Sanches ◽

...

Keyword(s):

Oxidative Stress ◽

Ascorbic Acid ◽

Culture Medium ◽

Bos Taurus ◽

Follicular Development ◽

Ovarian Follicles ◽

Bos Taurus Indicus ◽

Bonferroni Test ◽

And Oxidative Stress

Summary The aim of this study was to evaluate the follicular development, morphological integrity, and oxidative stress of preantral ovarian follicles from Bos taurus indicus females grown in vitro with ascorbic acid. Ovaries (n = 20) from Bos taurus indicus females were collected, fragmented, and were cultured in vitro for 6 or 12 days in minimum essential medium (MEM), or MEM supplemented with 50 or 100 ng/ml ascorbic acid, with an extracellular matrix of agarose gel, in an incubator at 38.5°C; every 2 days, 100% of the culture medium was replaced. The data were analyzed using the chi-squared test and/or Fisher’s exact test. In the event of a significant effect, the proportions were compared using a 2 × 2 proportion test. The oxidative stress analysis data were submitted to analysis of variance followed by the Bonferroni test. Values were considered significant when P ≤ 0.05. The addition of 100 ng/ml of ascorbic acid to the in vitro culture medium of preantral ovarian follicles from bovine females promoted follicular development, was efficient in maintaining morphological integrity, as well as the stability of reactive oxygen species, after 6 days of in vitro culture.

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Survival factors regulating ovarian apoptosis -- dependence on follicle differentiation

Reproduction ◽

10.1530/rep.0.1230023 ◽

2002 ◽

pp. 23-30 ◽

Cited By ~ 150

Author(s):

E Markstrom ◽

ECh Svensson ◽

R Shao ◽

B Svanberg ◽

H Billig

Keyword(s):

Growth Factor ◽

Follicular Development ◽

Antral Follicle ◽

Short Review ◽

Ovarian Follicles ◽

Survival Factors ◽

Primordial Follicles ◽

Survival Factor ◽

Fetal Ovary

Only a minute fraction of the ovarian follicles present in a fetal ovary will complete the path to ovulation. Most of the follicles will undergo atresia, a hormonally controlled apoptotic process. Apoptosis occurs at each stage of follicular development and there is a marked reduction in the number of follicles present at birth. Stage-dependent mechanisms of follicle survival can be postulated to achieve co-ordinated development, leading to ovulation of a small fraction of follicles. Indeed, hormone and growth factor regulation of follicular atresia is stage-specific. This short review considers the factors that promote survival of ovarian follicles throughout development, including endocrine, locally produced and intracellular mediators, as exemplified mainly by follicular development in rodents. In primordial follicles, oocyte apoptosis is considered to be the cause of subsequent follicle degeneration. In slow-growing preantral follicles, FSH is not a survival factor, but some locally produced growth factors are. Progression to the antral follicle stage is probably the most critical stage of follicle development in vivo, and FSH is a major survival factor at this stage. In addition, insulin-like growth factor I and interleukin 1beta are potent survival factors for cultured rat follicles at the antral stage. Preovulatory follicles express receptors for LH, and both of the gonadotrophins are survival factors at this stage. Relatively little is known about the period between the LH surge and ovulation; however, it has been suggested that at this stage progesterone acts as a survival factor.

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Development and survivability of ovarian follicles of goat in different feeding systems

Bangladesh Journal of Animal Science ◽

10.3329/bjas.v43i3.21644 ◽

2014 ◽

Vol 43 (3) ◽

pp. 175-179

Author(s):

MH Alam ◽

ME Kabir ◽

MB Sarker ◽

BK Saha ◽

RI Khan ◽

...

Keyword(s):

Histological Examination ◽

Morphometric Analysis ◽

Follicular Development ◽

Ovarian Follicles ◽

Primordial Follicles ◽

Antral Follicles ◽

Feeding Systems

The present study was aimed to know the effects of feeding systems on follicular development in Black Bengal goats. Nine female post-weaned Black Bengal goats were randomly assigned to 3 feeding systems namely stall feeding, tethering and grazing. After 240 days of rearing, goats were slaughtered and ovaries were collected for morphometric analysis. Histological examination revealed that the percentages of secondary (p<0.01) and antral follicles (p<0.05) were higher in stall-fed than tethering and grazing goats. Percentage of primordial follicles (61±2%) was lower in stall-fed goats and higher in tethering (71±3%) goats. Number of degenerated follicles was higher in tethering goats and lower in stall-fed groups. These results reveal that development and degeneration of ovarian follicles are influenced by feeding systems of goats.DOI: http://dx.doi.org/10.3329/bjas.v43i3.21644 Bang. J. Anim. Sci. 2014. 43 (3): 175-179

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153. XENOBIOTICS; INFLUENCE ON OVARIAN FOLLICULAR DEVELOPMENT

Reproduction Fertility and Development ◽

10.1071/srb09abs153 ◽

2009 ◽

Vol 21 (9) ◽

pp. 71

Author(s):

A. P. Sobinoff ◽

V. Pye ◽

B. Nixon ◽

S. D. Roman ◽

E. A. McLaughlin

Keyword(s):

Differentially Expressed Genes ◽

Molecular Mechanisms ◽

Follicular Development ◽

Ovarian Follicles ◽

Differentially Expressed ◽

Signalling Pathways ◽

Free Oxygen Radicals ◽

Toxic Response ◽

Primordial Follicles ◽

Significant Difference

The mammalian female reproductive lifespan is largely defined by a finite pool of ovarian follicles established around the time of birth. It is now understood that certain synthetic chemical compounds, known as xenobiotics, can cause premature ovarian senescence through the destruction of small ovarian follicles. Although the ovotoxic effects of these chemicals are well documented, the exact molecular mechanisms behind their action are only just becoming understood. Recent evidence suggests that bioactivation of xenobiotics by Phase I detoxifying enzymes may lead to the generation of free oxygen radicals (ROS), which we suspect may perturb intracellular signalling pathways in primordial follicles. In this study we attempted to identify ovarian follicle signalling pathways activated by xenobiotic exposure using ovotoxic agents which target immature follicles. Neonatal ovaries obtained from 3/4-day old Swiss mice were exposed to either 4-Vinylcyclohexene (25µM), Methoxychlor (25µM) or Menadione (5µM) for 96hrs using our in vitro culture system. Total RNA was then collected and analysed using Affymetrix Mouse Genome 430 2.0 Arrays. Bioinformatic analysis identified between ~500–1000 genes with a two-fold significant difference in gene expression (p<0.05) for each xenobiotic compared to the control. Differentially expressed genes were analysed for pathways and molecular functions using Ingenuity Pathways Analysis (Ingenuity Systems). In agreement with the current literature, many of the genes belonged to toxic response pathways, such as; Xenobiotic metabolism (10); p53 (15) and Apoptosis (11) signalling. However, the vast majority of the differentially expressed genes belonged to canonical pathways implicated in follicular development, such as PI3K/AKT (18), Wnt/ b -catenin (21), and JAK/Stat (8) signalling. Further qPCR analysis has confirmed a substantial increase in the transcription factor Sox4 and cell cycle inhibitor Cdkn2a in 4-Vinylcyclohexene and Menadione treated ovaries respectively. These results suggest that xenobiotics which target primordial follicles may exert part of their ovotoxic effects by perturbing signalling pathways involved in follicular activation and development.

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Antioxidant supplementation partially rescues accelerated ovarian follicle loss, but not oocyte quality, of glutathione-deficient mice†

Biology of Reproduction ◽

10.1093/biolre/ioaa009 ◽

2020 ◽

Vol 102 (5) ◽

pp. 1065-1079

Author(s):

Jinhwan Lim ◽

Samiha Ali ◽

Lisa S Liao ◽

Emily S Nguyen ◽

Laura Ortiz ◽

...

Keyword(s):

Oxidative Stress ◽

Lipoic Acid ◽

Ovarian Follicle ◽

Primordial Follicle ◽

Oocyte Quality ◽

Embryonic Mortality ◽

Wild Type ◽

Primordial Follicles ◽

Age Related ◽

Acetyl Cysteine

Abstract The tripeptide thiol antioxidant glutathione (GSH) has multiple physiological functions. Female mice lacking the modifier subunit of glutamate cysteine ligase (GCLM), the rate-limiting enzyme in GSH synthesis, have decreased GSH concentrations, ovarian oxidative stress, preimplantation embryonic mortality, and accelerated age-related decline in ovarian follicles. We hypothesized that supplementation with thiol antioxidants, N-acetyl cysteine (NAC), or α-lipoic acid (ALA) will rescue this phenotype. Gclm−/− and Gclm+/+ females received 0 or 80 mM NAC in drinking water from postnatal day (PND) 21–30; follicle growth was induced with equine chorionic gonadotropin (eCG) on PND 27, followed by an ovulatory dose of human CG and mating with a wild type male on PND 29 and zygote harvest 20 h after hCG. N-acetyl cysteine supplementation failed to rescue the low rate of second pronucleus formation in zygotes from Gclm−/− versus Gclm+/+ females. In the second study, Gclm−/− and Gclm+/+ females received diet containing 0, 150, or 600 mg/kg ALA beginning at weaning and were mated with wild type males from 8 to 20 weeks of age. α-Lipoic acid failed to rescue the decreased offspring production of Gclm−/− females. However, 150 mg/kg diet ALA partially rescued the accelerated decline in primordial follicles, as well as the increased recruitment of follicles into the growing pool and the increased percentages of follicles with γH2AX positive oocytes or granulosa cells of Gclm−/− females. We conclude that ovarian oxidative stress is the cause of accelerated primordial follicle decline, while GSH deficiency per se may be responsible for preimplantation embryonic mortality in Gclm−/− females.

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Age-Related Changes in Activation of Mitogen-Activated Protein Kinase Cascades by Oxidative Stress

Journal of Investigative Dermatology ◽

10.1038/jidsp.1998.7 ◽

1998 ◽

Vol 3 (1) ◽

pp. 23-27 ◽

Cited By ~ 9

Author(s):

Kathryn Z Guyton ◽

Myriani Gorospe ◽

Xiantao Wang ◽

Yolanda D Mock ◽

Gertrude C Kokkonen ◽

...

Keyword(s):

Oxidative Stress ◽

Protein Kinase ◽

Mitogen Activated Protein Kinase ◽

Age Related ◽

Mitogen Activated Protein ◽

Age Related Changes

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Serum of 8-OHdG as a potential biomarker of oxidative DNA damage in workers exposed to harmful working environments

Russian Journal of Occupational Health and Industrial Ecology ◽

10.31089/1026-9428-2019-59-9-783-784 ◽

2020 ◽

pp. 783-783

Author(s):

I. A. Umnyagina ◽

L. A. Strakhova ◽

T. V. Blinova

Keyword(s):

Oxidative Stress ◽

Dna Damage ◽

Blood Serum ◽

Oxidative Dna Damage ◽

Working Environment ◽

Working Environments ◽

Potential Biomarker ◽

High Level ◽

Damage Marker

In the blood serum of 70% individuals exposed to harmful factors of the working environment, a high level of oxidative stress and the DNA damage marker 8-Hydroxy-2’-Deoxyguanosine (8-OHdG) were detected.

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Oxidative Stress, Ocular Disease and Diabetes Retinopathy

Current Pharmaceutical Design ◽

10.2174/1381612825666190115121531 ◽

2019 ◽

Vol 24 (40) ◽

pp. 4726-4741 ◽

Cited By ~ 8

Author(s):

Orathai Tangvarasittichai ◽

Surapon Tangvarasittichai

Keyword(s):

Oxidative Stress ◽

Dna Damage ◽

Cell Death ◽

Protein Modification ◽

Morphological Changes ◽

Corneal Dystrophy ◽

Age Related Macular Degeneration ◽

Ocular Diseases ◽

Retinal Light Damage ◽

Age Related

Background: Oxidative stress is caused by free radicals or oxidant productions, including lipid peroxidation, protein modification, DNA damage and apoptosis or cell death and results in cellular degeneration and neurodegeneration from damage to macromolecules. Results: Accumulation of the DNA damage (8HOdG) products and the end products of LPO (including aldehyde, diene, triene conjugates and Schiff’s bases) were noted in the research studies. Significantly higher levels of these products in comparison with the controls were observed. Oxidative stress induced changes to ocular cells and tissues. Typical changes include ECM accumulation, cell dysfunction, cell death, advanced senescence, disarrangement or rearrangement of the cytoskeleton and released inflammatory cytokines. It is involved in ocular diseases, including keratoconus, Fuchs endothelial corneal dystrophy, and granular corneal dystrophy type 2, cataract, age-related macular degeneration, primary open-angle glaucoma, retinal light damage, and retinopathy of prematurity. These ocular diseases are the cause of irreversible blindness worldwide. Conclusions: Oxidative stress, inflammation and autophagy are implicated in biochemical and morphological changes in these ocular tissues. The development of therapy is a major target for the management care of these ocular diseases.

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Kisspeptin level in the aging ovary is regulated by the sympathetic nervous system

Journal of Endocrinology ◽

10.1530/joe-16-0181 ◽

2017 ◽

Vol 232 (1) ◽

pp. 97-105 ◽

Cited By ~ 12

Author(s):

Daniela Fernandois ◽

Gonzalo Cruz ◽

Eun Kyung Na ◽

Hernán E Lara ◽

Alfonso H Paredes

Keyword(s):

Adrenergic Receptor ◽

Follicular Development ◽

Sympathetic Nerves ◽

Female Rats ◽

Reproductive Aging ◽

Beta Adrenergic Receptor ◽

Beta Adrenergic ◽

Primordial Follicles ◽

Follicular Cyst

Previous work has demonstrated that the increase in the activity of sympathetic nerves, which occurs during the subfertility period in female rats, causes an increase in follicular cyst development and impairs follicular development. In addition, the increase in ovarian sympathetic activity of aged rats correlates with an increased expression of kisspeptin (KISS1) in the ovary. This increase in KISS1 could participate in the decrease in follicular development that occurs during the subfertility period. We aimed to determine whether the blockade of ovarian sympathetic tone prevents the increase in KISS1 expression during reproductive aging and improves follicular development. We performed 2 experiments in rats: (1) an in vivo blockade of beta-adrenergic receptor with propranolol (5.0 mg/kg) and (2) an ovarian surgical denervation to modulate the sympathetic system at these ages. We measured Kisspeptin and follicle-stimulating hormone receptor (FSHR) mRNA and protein levels by qRT-PCR and western blot and counted primordial, primary and secondary follicles at 8, 10 and 12 months of age. The results showed that ovarian KISS1 decreased but FSHR increased after both propranolol administration and the surgical denervation in rats of 8, 10 and 12 months of age. An increase in FSHR was related to an increase in the number of smaller secondary follicles and a decreased number of primordial follicles at 8, 10 and 12 months of age. These results suggest that intraovarian KISS1 is regulated by sympathetic nerves via a beta-adrenergic receptor and participates locally in ovarian follicular development in reproductive aging.

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Zone-specific damage of the olfactory epithelium under protein restriction

Scientific Reports ◽

10.1038/s41598-020-79249-3 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Ayinuer Tuerdi ◽

Shu Kikuta ◽

Makoto Kinoshita ◽

Teru Kamogashira ◽

Kenji Kondo ◽

...

Keyword(s):

Oxidative Stress ◽

Olfactory Epithelium ◽

Control Diet ◽

Intervention Strategy ◽

Protein Restriction ◽

Outer Hair Cells ◽

Basal Cells ◽

Cell Dynamics ◽

Age Related ◽

Positive Effect

AbstractOxidative stress causes tissue damage, affecting age-related pathologies. Protein restriction (PR) provides a powerful intervention strategy for reducing oxidative stress, which may have a positive effect on individual organs. However, it is unknown whether PR intervention influences the olfactory system. Here, we investigated how 10 months of PR could affect the cell dynamics of the olfactory epithelium (OE) in mice. We found that PR reduced age-related loss of outer hair cells in the cochlea, providing preventive effects against age-related hearing loss. In contrast, PR resulted in reduced mature olfactory sensory neurons (OSNs), increased proliferative basal cells, and increased apoptotic OSNs in zone 1 (the only area containing neurons expressing NQO1 [quinone dehydrogenase 1]) of the OE in comparison with animals given a control diet. Substantial oxidative stress occurred in NQO1-positive cells and induced apoptotic OSNs in zone 1. These results indicate that in contrast to the positive effect on the auditory system, PR induces oxidative stress and structurally and functionally negative effects on OSNs in zone 1, which is probably involved in the bioactivation of NQO1.

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