AOP: 492

Title

A descriptive phrase which references both the Molecular Initiating Event and Adverse Outcome.It should take the form “MIE leading to AO”. For example, “Aromatase inhibition leading to reproductive dysfunction” where Aromatase inhibition is the MIE and reproductive dysfunction the AO. In cases where the MIE is unknown or undefined, the earliest known KE in the chain (i.e., furthest upstream) should be used in lieu of the MIE and it should be made clear that the stated event is a KE and not the MIE. More help

Glutathione conjugation leading to reproductive dysfunction via oxidative stress

Short name

A name that succinctly summarises the information from the title. This name should not exceed 90 characters. More help

Glutathione conjugation leading to reproductive dysfunction

The current version of the Developer's Handbook will be automatically populated into the Handbook Version field when a new AOP page is created.Authors have the option to switch to a newer (but not older) Handbook version any time thereafter. More help

Handbook Version v2.5

Graphical Representation

A graphical representation of the AOP.This graphic should list all KEs in sequence, including the MIE (if known) and AO, and the pair-wise relationships (links or KERs) between those KEs. More help

Click to download graphical representation template Explore AOP in a Third Party Tool

Authors

The names and affiliations of the individual(s)/organisation(s) that created/developed the AOP. More help

Vieira, Leonardo

Farias, Davi

Souza, Terezinha

Point of Contact

The user responsible for managing the AOP entry in the AOP-KB and controlling write access to the page by defining the contributors as described in the next section. More help

Allie Always (email point of contact)

Contributors

Users with write access to the AOP page. Entries in this field are controlled by the Point of Contact. More help

Leonardo Vieira
Allie Always

Coaches

This field is used to identify coaches who supported the development of the AOP.Each coach selected must be a registered author. More help

OECD Information Table

Provides users with information concerning how actively the AOP page is being developed and whether it is part of the OECD Workplan and has been reviewed and/or endorsed. OECD Project: Assigned upon acceptance onto OECD workplan. This project ID is managed and updated (if needed) by the OECD. OECD Status: For AOPs included on the OECD workplan, ‘OECD status’ tracks the level of review/endorsement of the AOP . This designation is managed and updated by the OECD. Journal-format Article: The OECD is developing co-operation with Scientific Journals for the review and publication of AOPs, via the signature of a Memorandum of Understanding. When the scientific review of an AOP is conducted by these Journals, the journal review panel will review the content of the Wiki. In addition, the Journal may ask the AOP authors to develop a separate manuscript (i.e. Journal Format Article) using a format determined by the Journal for Journal publication. In that case, the journal review panel will be required to review both the Wiki content and the Journal Format Article. The Journal will publish the AOP reviewed through the Journal Format Article. OECD iLibrary published version: OECD iLibrary is the online library of the OECD. The version of the AOP that is published there has been endorsed by the OECD. The purpose of publication on iLibrary is to provide a stable version over time, i.e. the version which has been reviewed and revised based on the outcome of the review. AOPs are viewed as living documents and may continue to evolve on the AOP-Wiki after their OECD endorsement and publication. More help

OECD Project #	OECD Status	Reviewer's Reports	Journal-format Article	OECD iLibrary Published Version

This AOP was last modified on May 26, 2024 20:39

Revision dates for related pages

Page	Revision Date/Time
Depletion, GSH	September 14, 2023 10:54
Conjugation, GSH	September 14, 2023 10:47
Increased, Reactive oxygen species	April 10, 2024 17:33
Increased, Lipid peroxidation	July 27, 2023 10:25
impaired, Fertility	September 14, 2023 12:10
Conjugation, GSH leads to Depletion, GSH	September 14, 2023 12:38
Depletion, GSH leads to Increased, Reactive oxygen species	September 14, 2023 12:46
Increased, Reactive oxygen species leads to Increased, LPO	April 11, 2024 16:24
Increased, LPO leads to impaired, Fertility	September 14, 2023 13:02
atrazine	November 29, 2016 18:42
Mercuric chloride	November 29, 2016 18:42
Diethyl maleate	April 11, 2023 13:03

Abstract

A concise and informative summation of the AOP under development that can stand-alone from the AOP page. The aim is to capture the highlights of the AOP and its potential scientific and regulatory relevance. More help

Here, an Adverse Outcome Pathway (AOP) is proposed for reproductive dysfunction via oxidative stress, which is motivated by the current understanding of the role of oxidative stress in reproductive disorders. The AOP was developed based on OECD's guide no. 184 and the specific considerations of OECD Users' handbook supplement to the guidance document for developing and assessing AOPs (no. 233).

According to qualitative and quantitative experimental data that were evaluated, GSH conjugation is the first upstream Key Event (KE) of this AOP, triggering oxidative stress (OS). This event causes depletion of GSH basal levels (KE2). Consequently, this reduction of free GSH induces an increase of ROS (KE3) generated by natural cellular metabolic processes (cellular respiration) of the organisms. As expected, the intensified growth of these reactive species' levels, in turn, induces an increase of lipid peroxidation (KE4). This KE, consequently, leads to a rise in the amount of toxic substances, such as malondialdehyde and hydroxynonenal. Both are intrinsically associated with the decrease in the quality and competence of gamete cell division, and, consequently, cause impairment of fertility (KE5 and Adverse Outcome).

AOP Development Strategy

Context

Used to provide background information for AOP reviewers and users that is considered helpful in understanding the biology underlying the AOP and the motivation for its development.The background should NOT provide an overview of the AOP, its KEs or KERs, which are captured in more detail below. More help

This AOP was developed for the project "CHRONIC TOXICITY OF PESTICIDES IN DRINKING WATER IN PARAÍBA (TRIGGER): IDENTIFYING THE TRIGGERS OF A SILENT EPIDEMIC," financed by the "Fundação de Apoio à Pesquisa do Estado da Paraíba (FAPESQ-PB)." The project aims to understand how oxidative stress and reproductive toxicity can be triggered in animals by aquatic pollutants, such as atrazine

Strategy

Provides a description of the approaches to the identification, screening and quality assessment of the data relevant to identification of the key events and key event relationships included in the AOP or AOP network.This information is important as a basis to support the objective/envisaged application of the AOP by the regulatory community and to facilitate the reuse of its components. Suggested content includes a rationale for and description of the scope and focus of the data search and identification strategy/ies including the nature of preliminary scoping and/or expert input, the overall literature screening strategy and more focused literature surveys to identify additional information (including e.g., key search terms, databases and time period searched, any tools used). More help

The first step in developing this AOP was to conduct a literature review to gather toxicological data on the herbicide atrazine's impact on aquatic vertebrates in relation to oxidative stress and reproductive toxicity.

We used keywords such as "atrazine," "fish," "oxidative stress," and "reproductive toxicity" to search databases like ScienceDirect and PubMed. Screening of studies was based on analyzing titles and abstracts, and only articles published in indexed journals and written in English were considered.

Upon analyzing the works, evidence was found indicating that one of the primary detoxification mechanisms of atrazine in fish involved its conjugation with reduced glutathione (GSH), and a direct relationship was observed between oxidative damage and reproductive toxicity caused by the herbicide in this taxonomic group.

Subsequently, we conducted another review using the keywords "oxidative stress" and "infertility" in specialized literature and confirmed a close relationship between oxidative stress and infertility not only in fish but also in mammals and birds. Furthermore, it was noted that GSH plays a crucial role in ensuring the reproductive success of animals, including humans.

In this context, impaired fertility and GSH conjugation were hypothesized as Adverse Outcome (AO) and Molecular Initiating Event (MIE), respectively, for this AOP. Subsequently, increased ROS and increased lipid peroxidation were selected as key intermediate events.

Finally, to assess the weight of evidence for the AOP, we conducted a literature review to include toxicological data not only for atrazine but also for mercury and diethyl maleate (as they activate AO through the same mechanism), this time without taxon restrictions. Additionally, for quantitative understanding, data on response-response relationships from pairs of adjacent KEs were utilized.

Summary of the AOP

This section is for information that describes the overall AOP.The information described in section 1 is entered on the upper portion of an AOP page within the AOP-Wiki. This is where some background information may be provided, the structure of the AOP is described, and the KEs and KERs are listed. More help

Events:

Molecular Initiating Events (MIE)

An MIE is a specialised KE that represents the beginning (point of interaction between a prototypical stressor and the biological system) of an AOP. More help

Key Events (KE)

A measurable event within a specific biological level of organisation. More help

Adverse Outcomes (AO)

An AO is a specialized KE that represents the end (an adverse outcome of regulatory significance) of an AOP. More help

Type	Event ID	Title	Short name

MIE

2131

Conjugation, GSH

KE	130	Depletion, GSH	Depletion, GSH
KE	1115	Increased, Reactive oxygen species	Increased, Reactive oxygen species
KE	1445	Increased, Lipid peroxidation	Increased, LPO

406

impaired, Fertility

Relationships Between Two Key Events (Including MIEs and AOs)

This table summarizes all of the KERs of the AOP and is populated in the AOP-Wiki as KERs are added to the AOP.Each table entry acts as a link to the individual KER description page. More help

Title	Adjacency	Evidence	Quantitative Understanding

Conjugation, GSH leads to Depletion, GSH	adjacent	High	High
Depletion, GSH leads to Increased, Reactive oxygen species	adjacent	High	High
Increased, Reactive oxygen species leads to Increased, LPO	adjacent	High	High
Increased, LPO leads to impaired, Fertility	adjacent	High	High

Network View

This network graphic is automatically generated based on the information provided in the MIE(s), KEs, AO(s), KERs and Weight of Evidence (WoE) summary tables. The width of the edges representing the KERs is determined by its WoE confidence level, with thicker lines representing higher degrees of confidence. This network view also shows which KEs are shared with other AOPs. More help

Prototypical Stressors

A structured data field that can be used to identify one or more “prototypical” stressors that act through this AOP. Prototypical stressors are stressors for which responses at multiple key events have been well documented. More help

Name
atrazine
Mercuric chloride
Diethyl maleate

Life Stage Applicability

The life stage for which the AOP is known to be applicable. More help

Life stage	Evidence
Adults	High

Taxonomic Applicability

Latin or common names of a species or broader taxonomic grouping (e.g., class, order, family) can be selected.In many cases, individual species identified in these structured fields will be those for which the strongest evidence used in constructing the AOP was available. More help

Term	Scientific Term	Evidence	Link
mammals	mammals	High	NCBI
fish	fish	High	NCBI

Sex Applicability

The sex for which the AOP is known to be applicable. More help

Sex	Evidence
Unspecific	High

Overall Assessment of the AOP

Addressess the relevant biological domain of applicability (i.e., in terms of taxa, sex, life stage, etc.) and Weight of Evidence (WoE) for the overall AOP as a basis to consider appropriate regulatory application (e.g., priority setting, testing strategies or risk assessment). More help

Biological plausibility, empirical support and quantitative understanding of the KERs and the evidence that uphold essentialities of KEs in this AOP were analyzed together for the overall assessment of an AOP. In this case, overall assessment (WoE) of the general biological plausibility and of the empirical support of KERs was considered as high for this AOP, as well as essentiality, once for this criterion the first four KEs that trigger the AO are also classified as such. Finally, although the amount of data that support each of the relations differed considerably among them in number, it was possible to obtain an overview about the quantitative comprehension of the KERs, as well as understand their mechanisms. Nevertheless, it is suitable to suggest that more data must be generated, with regard to KER 2879, in order to improve comprehension of this relation among different taxonomic groups.

Domain of Applicability

Addressess the relevant biological domain(s) of applicability in terms of sex, life-stage, taxa, and other aspects of biological context. More help

Empirical domain of applicability

Sex: The AOP is applicable to males and females.

Life stages: All life stages are relevant to this AOP.

Taxonomic: The assumed empirical domain of applicability of this AOP is fish and mammals.

Biologically plausible domain of applicability

All the key events described should be conserved among animal species, suggesting that the AOP may also have relevance for amphibians, reptiles, birds and and invertebrates with sexual reproduction. However, interspecies differences are possible because the effectiveness of GSH conjugation as a detoxification mechanism may depend on the species and the specific chemical being considered (Summer et al., 1979).

Essentiality of the Key Events

The essentiality of KEs can only be assessed relative to the impact of manipulation of a given KE (e.g., experimentally blocking or exacerbating the event) on the downstream sequence of KEs defined for the AOP. Consequently, evidence supporting essentiality is assembled on the AOP page, rather than on the independent KE pages that are meant to stand-alone as modular units without reference to other KEs in the sequence. The nature of experimental evidence that is relevant to assessing essentiality relates to the impact on downstream KEs and the AO if upstream KEs are prevented or modified. This includes: Direct evidence: directly measured experimental support that blocking or preventing a KE prevents or impacts downstream KEs in the pathway in the expected fashion. Indirect evidence: evidence that modulation or attenuation in the magnitude of impact on a specific KE (increased effect or decreased effect) is associated with corresponding changes (increases or decreases) in the magnitude or frequency of one or more downstream KEs. More help

After blocking the synthesis of GSH with the inhibitor buthionine sulfoximine (BSO) – at a dose of 2 mmol/kg at 12-hour intervals for 7 days – male rats (4 months old) experienced a dramatic decrease in GSH levels. In the seminal vesicles, there was a depletion of 71% in the content, while in the epididymal tissues, this depletion was more severe: 81% in the caput, 87% in the corpus, and 92% in the cauda of the epididymis. Furthermore, the enzymatic activity of catalase increased significantly in the epididymal tissues, while, on the other hand, the activity of manganese superoxide dismutase (Mn SOD) and glutathione peroxidase (GPX) decreased in the seminal vesicle. Additionally, the sperm motility of the animals was reduced (Zubkova et al., 2004).

In another in vivo study, the administration of BSO for 35 days in BALB/c mice at 8 weeks of age – at 2 mmol/kg/day – caused a decrease in GSH content, as well as in catalase (CAT), SOD, and GPX activity. Meanwhile, the MDA content in the testes increased considerably, and a reduction in fertility was recorded through a decrease in normal sperm and sperm motility and an increase in abnormal sperm (Sajjadian et al., 2014). Moreover, according to Lopez and Luderer (2004), rats treated with BSO 5 mmol/kg body weight twice a day showed both a decrease in GSH content and an increase in atretic antral follicles in the ovaries. On the other hand, rats treated with BSO 4 mmol/kg of body weight twice a day showed significantly decreased levels of GSH and enzymatic activity of CAT, SOD, and GPX in blood and erythrocytes, as well as increased levels of MDA. However, glutathione-monoester therapy during exposure promoted the recovery of levels and activity of these oxidative stress markers in animals treated with BSO (Rajasekaran et al., 2004).

In male Nrf2^-/- knockout mice, there was a reduction in gene expression levels of antioxidant enzymes in the testis and epididymis, including catalytic glutamate cysteine ligase (Gclc), glutamate cysteine ligase modifying subunit (Gclm) – the rate-limiting enzyme in GSH synthesis – glutathione transferase m1 (Gstm1), Gstm2, Gsta3, and Sod2, as well as a depletion in GSH concentration and GPX activity compared to wild-type males. In addition, MDA levels were shown to be significantly increased, while fertility was reduced by the decrease in the number of litters and pups (Nakamura et al., 2010). Furthermore, Nakamura et al. (2011) showed that Gclm null female mice show a decrease in GSH content in ovulated oocytes and a decrease in fertility through the reduction of litter and offspring production. Additionally, Lim et al. (2015) found a drop in GSH levels and Nernst potential (Eh) (indicating oxidative stress), an increase in 4-hydroxynonenal (4-HNE), and a decline in ovarian follicles in Gclm null female mice. Besides this, Lim et al. (2020) showed that female mice lacking the Gclm gene show depleted GSH concentrations and a reduction in the number of healthy follicles.

Moreover, Garratt et al. (2013) showed that Sod1^-/- mice have impaired sperm motility and in vivo fertilization compared to WT animals. Furthermore, Imai et al. (2009) showed that spermatocyte-specific Gpx4^-/- knockout mice are completely infertile, whereas GPx4^+/− and transgenic rescued Gpx4^-/-knockout mice were fully fertile. Additionally, according to Schneider et al. (2009), mGpx4^-/- (mitochondrial GPx4) knockout mice are infertile and have less motile and progressive sperm compared to WT.

Table 2: Summary of in vivo studies with fertility endpoints for chemical inhibitors or gene knockout experiments as evidence to support the essentiality of KEs.

Study	Treatment	GSH	ROS	Lipid peroxidation	Fertility
Zubkova et al., 2004	2 mmol/kg BSO 7 d rat (Young)	↓content	↑CAT, total SOD, Mn SOD and GPx activity	−	↓via spermatozoal motility
Zubkova et al., 2004	2 mmol/kg BSO 7 d rat (Old)	↓content	↑via CAT activity	−	↓via spermatozoal motility
Sajjadian et al., 2014	2 mmol/kg/day BSO 35 d mice	↓content	↑ via CAT, GPx and SOD units	↑ via MDA	↓via sperm motility and increase of abnormal sperms
Lopez and Luderer, 2004	5 mmol/kg BSO 24 h rat	↓content	−	−	↓via atretic antral follicles
Nakamura et al., 2010	Nrf2^-/- knockout mice	↓content	↑ via Gclc, Gclm, Gstm1, Gstm2, Gsta3 and SOD2 gene expression and GPx units	↑ via MDA and HAE*	↓via sperm counts, sperm motility, litters and offspring
Nakamura et al. 2011	Gclm^-/- null mice	↓content	−	−	↓via litter and offspring
Lim et al. 2015	Gclm^-/- null mice	↓content	↑ via Nernst potential (E_h)	↑ via 4-HNE	↓via ovarian follicles
Lim et al. 2020	Gclm^-/- null mice	↓content	−		↓via healthy follicles
Garratt et al. 2013	Sod1^-/- knockout mice	−	−	−	↓via sperm motility, fertility rates
Schneider et al. 2009	mGPx ^-/-knockout mice	−	−	−	↓via sperm motility and litter
Imai et al. 2009	mGPx ^-/-knockout mice	−	−	−	↓via sperm count, motility, fertility rates

Evidence Assessment

Addressess the biological plausibility, empirical support, and quantitative understanding from each KER in an AOP. More help

Several chemicals that undergo GSH conjugation at high concentrations cause depletion of GSH supplies in the liver and other tissues (D’Souza, Francis, and Andersen 1988; D’Souza and Andersen 1988; Csanády et al. 1996; Mulder and Ouwerkerk-Mahadevan 1997; Fennell and Brown 2001).

Diethyl maleate at 0.1, 0.5, 1, 2.5, and 5 mM for five hours caused GSH depletion in hepatocytes at all concentrations in a dose-dependent manner. However, only 5 mM of the compound was able to consume GSH to the point that this antioxidant was kept below detection levels (4%) and led to overproduction of ROS (Tirmenstein et al. 2000).

Adult rats treated with BSO 20 and 30 mM for 10 days diligently showed a reduction of, respectively, 44.25% and 60.14% of liver GSH content, while H2O2 levels underwent an augmentation of 42 and 60%, in that order (Ford et al. 2006).

For instance, empirical evidence shows that rat hepatocytes begin ROS production after the first 30 minutes of DEM exposition (5 mM), growing linearly for all the remaining time, whereas the increase in products of lipid peroxidation (TBARS) starts only from the first hour of exposure (Tirmenstein et al. 2000).

Experimental evidence showed that the lipid peroxidation product 4-HNE, at 0, 5, 10, 20, 30, and 50 µM, induces a dose-dependent decrease in meiotic competence during in vitro oocyte maturation, as well as aneuploidies in germinal vesicle (GV) oocytes from 20 µM of 4-HNE (Mihalas et al. 2017).

BSO for 35 days in BALB/c mice at 8 weeks of age – at 2 mmol/kg/day – caused a decrease in GSH content, as well as in catalase (CAT), SOD, and GPX activity. Meanwhile, the MDA content in the testes increased considerably, and reduction in fertility was recorded through a decrease in normal sperm and sperm motility and an increase in abnormal sperm (Sajjadian et al., 2014).

Lim et al. (2015) found a drop in GSH levels and Nernst potential (Eh) (indicating oxidative stress), an increase in 4-hydroxynonenal (4-HNE), and a decline in ovarian follicles in Gclm null female mice.

Known Modulating Factors

Modulating factors (MFs) may alter the shape of the response-response function that describes the quantitative relationship between two KES, thus having an impact on the progression of the pathway or the severity of the AO.The evidence supporting the influence of various modulating factors is assembled within the individual KERs. More help

Modulating Factor (MF)	Influence or Outcome	KER(s) involved
Biflavonone-kolaviron	prevent GSH depletion	KER 2877
vitamin E	prevent GSH depletion	KER 2877
vitamin E	restores the activity of antioxidant enzymes	KER 2878
vitamin E	prevent lipid peroxidation	KER 2460
vitamin C	prevent lipid peroxidation	KER 2460

Quantitative Understanding

Optional field to provide quantitative weight of evidence descriptors. More help

Considerations for Potential Applications of the AOP (optional)

Addressess potential applications of an AOP to support regulatory decision-making.This may include, for example, possible utility for test guideline development or refinement, development of integrated testing and assessment approaches, development of (Q)SARs / or chemical profilers to facilitate the grouping of chemicals for subsequent read-across, screening level hazard assessments or even risk assessment. More help

References

List of the literature that was cited for this AOP. More help

D’Souza, R. W., and M. E. Andersen. 1988. “Physiologically Based Pharmacokinetic Model for Vinylidene Chloride.” Toxicology and Applied Pharmacology 95 (2): 230–40.

D’Souza, R. W., W. R. Francis, and M. E. Andersen. 1988. “Physiological Model for Tissue Glutathione Depletion and Increased Resynthesis after Ethylene Dichloride Exposure.” The Journal of Pharmacology and Experimental Therapeutics 245 (2): 563–68.

Csanády, G. A., P. E. Kreuzer, C. Baur, and J. G. Filser. 1996. “A Physiological Toxicokinetic Model for 1,3-Butadiene in Rodents and Man: Blood Concentrations of 1,3-Butadiene, Its Metabolically Formed Epoxides, and of Haemoglobin Adducts--Relevance of Glutathione Depletion.” Toxicology 113 (1-3): 300–305.

Mulder, G. J., and S. Ouwerkerk-Mahadevan. 1997. “Modulation of Glutathione Conjugation in Vivo: How to Decrease Glutathione Conjugation in Vivo or in Intact Cellular Systems in Vitro.” Chemico-Biological Interactions 105 (1): 17–34.

Fennell, T. R., and C. D. Brown. 2001. “A Physiologically Based Pharmacokinetic Model for Ethylene Oxide in Mouse, Rat, and Human.” Toxicology and Applied Pharmacology 173 (3): 161–75.

Tirmenstein, M. A., F. A. Nicholls-Grzemski, J. G. Zhang, and M. W. Fariss. 2000. “Glutathione Depletion and the Production of Reactive Oxygen Species in Isolated Hepatocyte Suspensions.” Chemico-Biological Interactions 127 (3): 201–17.

Ford, Rebecca J., Drew A. Graham, Steven G. Denniss, Joe Quadrilatero, and James W. E. Rush. 2006. “Glutathione Depletion in Vivo Enhances Contraction and Attenuates Endothelium-Dependent Relaxation of Isolated Rat Aorta.” Free Radical Biology & Medicine 40 (4): 670–78.

Garratt, M., Bathgate, R., de Graaf, S. P., and Brooks, R. C. 2013. “Copper-zinc superoxide dismutase deficiency impairs sperm motility and in vivo fertility.” Reproduction, 146(4), 297-304.

Schneider, M., Forster, H., Boersma, A., Seiler, A., Wehnes, H., Sinowatz, F., ... and Conrad, M. 2009. “Mitochondrial glutathione peroxidase 4 disruption causes male infertility”. The FASEB journal, 23(9), 3233-3242.

Imai, H., Hakkaku, N., Iwamoto, R., Suzuki, J., Suzuki, T., Tajima, Y., ... and Nakagawa, Y. 2009. “Depletion of selenoprotein GPx4 in spermatocytes causes male infertility in mice”. Journal of Biological Chemistry, 284(47), 32522-32532.

Lim, J., Ali, S., Liao, L. S., Nguyen, E. S., Ortiz, L., Reshel, S., and Luderer, U. 2020. “Antioxidant supplementation partially rescues accelerated ovarian follicle loss, but not oocyte quality, of glutathione-deficient mice.” Biology of Reproduction, 102(5), 1065-1079.

Lim, J., Nakamura, B. N., Mohar, I., Kavanagh, T. J., and Luderer, U. 2015. “Glutamate cysteine ligase modifier subunit (Gclm) null mice have increased ovarian oxidative stress and accelerated age-related ovarian failure.” Endocrinology, 156(9), 3329-3343.

Nakamura, B. N., Lawson, G., Chan, J. Y., Banuelos, J., Cortés, M. M., Hoang, Y. D., ... and Luderer, U. 2010. “Knockout of the transcription factor NRF2 disrupts spermatogenesis in an age-dependent manner. Free Radical Biology and Medicine, 49(9), 1368-1379.

Nakamura, B. N., Fielder, T. J., Hoang, Y. D., Lim, J., McConnachie, L. A., Kavanagh, T. J., and Luderer, U. 2011. Lack of maternal glutamate cysteine ligase modifier subunit (Gclm) decreases oocyte glutathione concentrations and disrupts preimplantation development in mice.” Endocrinology, 152(7), 2806-2815.

Lopez, S. G., and Luderer, U. 2004. “Effects of cyclophosphamide and buthionine sulfoximine on ovarian glutathione and apoptosis.” Free Radical Biology and Medicine, 36(11), 1366-1377.

Sajjadian, F., Roshangar, L., Hemmati, A., Nori, M., Soleimani-Rad, S., and Soleimani-Rad, J. 2014. “The effect of BSO-induced oxidative stress on histologic feature of testis: testosterone secretion and semen parameters in mice.” Iranian journal of basic medical sciences, 17(8), 606.

Zubkova, E. V., and Robaire, B. 2004. “Effect of glutathione depletion on antioxidant enzymes in the epididymis, seminal vesicles, and liver and on spermatozoa motility in the aging brown Norway rat.” Biology of reproduction, 71(3), 1002-1008.

Rajasekaran, N. S., Devaraj, N. S., and Devaraj, H. 2004. “Modulation of rat erythrocyte antioxidant defense system by buthionine sulfoximine and its reversal by glutathione monoester therapy.’ Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, 1688(2), 121-129.

Summer, K. H., Rozman, K., Coulston, F., and Greim, H. 1979. Urinary excretion of mercapturic acids in chimpanzees and rats. Toxicology and Applied Pharmacology, 50(2), 207-212.