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Keywords = HPG-L axis

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18 pages, 4023 KiB  
Article
Lactobacillus brevis GKJOY Supplementation Ameliorates Oxidative Stress and Reproductive Dysfunction in Male Rats with Polystyrene Microplastics-Induced Reproductive Toxicity
by Yi-Yuh Hwang, Sabri Sudirman, Yu-Chen Hsu, Chin-Chu Chen, Fanbin Kong, Deng-Fwu Hwang and Zwe-Ling Kong
Int. J. Mol. Sci. 2025, 26(10), 4533; https://doi.org/10.3390/ijms26104533 - 9 May 2025
Viewed by 698
Abstract
The growing demand for plastic products has led to an increase in human exposure to microplastics (MPs). MPs have been shown to have detrimental effects on reproductive function, while probiotics have demonstrated promise in enhancing fertility. This study aimed to determine the protective [...] Read more.
The growing demand for plastic products has led to an increase in human exposure to microplastics (MPs). MPs have been shown to have detrimental effects on reproductive function, while probiotics have demonstrated promise in enhancing fertility. This study aimed to determine the protective effects of Lactobacillus brevis GKJOY against reproductive damage induced by polystyrene microplastics (PS-MPs) in male rats. In the cell study, LC540 cells were treated with L. brevis GKJOY postbiotic (PGK), gamma-aminobutyric acid (GABA), and PS-MPs to evaluate their effects on cell viability and reactive oxygen species (ROS) production. In the animal experiment, rats were treated with a low dose of L. brevis GKJOY (GK1X, 50 mg/kg), a medium dose (GK2X, 100 mg/kg), or a high dose (GK4X, 200 mg/kg). The results showed that PGK and GABA reduced the levels of ROS and protected against oxidative stress. In contrast, PS-MPs increased ROS levels and had harmful effects on cell viability. In the animal study, testicular injuries caused by PS-MPs led to disruption of the hypothalamic–pituitary–gonadal (HPG) axis and a decrease in reproductive hormone levels. However, treatment with L. brevis GKJOY reduced oxidative stress and pro-inflammatory cytokine levels, restored hormonal imbalances, and led to significant improvements. L. brevis GKJOY effectively mitigated reproductive damage in male rats due to its dual function as a probiotic and neurotransmitter modulator. In conclusion, L. brevis GKJOY, which functions as both a probiotic and a GABA producer, may offer superior protection against male reproductive damage. Full article
(This article belongs to the Section Molecular Pharmacology)
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15 pages, 15465 KiB  
Article
Functional Involvement of Melatonin and Its Receptors in Reproductive Regulation of the Marine Teleost, Large Yellow Croaker (Larimichthys crocea)
by Xudong Liang, Jixiu Wang, Baoyi Huang, Haojie Yuan, Yucheng Ren, Chenqian Wu, Tianming Wang and Jingwen Yang
Fishes 2025, 10(1), 28; https://doi.org/10.3390/fishes10010028 - 10 Jan 2025
Cited by 3 | Viewed by 1123
Abstract
Melatonin is a critical regulator of biological rhythms across organisms, transducing light signals into neuroendocrine signals that facilitate reproductive regulation in response to environmental cues. However, the precise mechanisms through which melatonin regulates reproduction in fish require further investigation. In this study, we [...] Read more.
Melatonin is a critical regulator of biological rhythms across organisms, transducing light signals into neuroendocrine signals that facilitate reproductive regulation in response to environmental cues. However, the precise mechanisms through which melatonin regulates reproduction in fish require further investigation. In this study, we employed molecular and organizational biological techniques to examine the expression patterns of melatonin and its five receptor subtypes (LcMTNR1A1, LcMTNR1A2, LcMTNR1B1, LcMTNR1B2, and LcMTNR1C) in various tissues of the large yellow croaker (Larimichthys crocea). Our results revealed significant expression of all receptors in the pituitary and testes, with distinct gender differences, including a lack of expression in the ovary. Moreover, our data indicate that melatonin and its receptors are primarily expressed during stage III, highlighting their role in sexual maturity. Enzyme- linked immunosorbent assay (ELISA) results further demonstrated that in vitro melatonin incubation in the brain of L. crocea influenced gonadotropin-releasing hormone (GnRH) and testosterone secretion in a dose-dependent manner, suggesting actions beyond the classical hypothalamic–pituitary–gonadal (HPG) axis. Overall, our findings provide new evidence supporting the role of the melatonin system in reproductive regulation in marine teleosts. Full article
(This article belongs to the Special Issue Rhythms in Marine Fish and Invertebrates)
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18 pages, 2503 KiB  
Article
Reproductive and Developmental Effects of Sex-Specific Chronic Exposure to Dietary Arsenic in Zebrafish (Danio rerio)
by Mahesh Rachamalla, Arash Salahinejad, Vladimir Kodzhahinchev and Som Niyogi
Toxics 2024, 12(4), 302; https://doi.org/10.3390/toxics12040302 - 19 Apr 2024
Cited by 5 | Viewed by 2545
Abstract
The present study investigated the reproductive and developmental effects of sex-specific chronic exposure to dietary arsenic in zebrafish. Adult zebrafish (Danio rerio) were exposed to environmentally realistic doses of arsenic via diet [0 (control; no added arsenic), 30 (low), 60 (medium), [...] Read more.
The present study investigated the reproductive and developmental effects of sex-specific chronic exposure to dietary arsenic in zebrafish. Adult zebrafish (Danio rerio) were exposed to environmentally realistic doses of arsenic via diet [0 (control; no added arsenic), 30 (low), 60 (medium), and 100 (high) μg/g dry weight, as arsenite] for 90 days. Following exposure, arsenic-exposed females from each dietary treatment were mated with control males, and similarly, arsenic-exposed males from each dietary treatment were mated with control females. In females, arsenic exposure resulted in a dose-dependent decrease in reproductive performance (fecundity, fertilization success, and hatching success). Moreover, a dose-dependent increase in developmental toxicity (larval deformities and larval mortality) was observed with maternal exposure to arsenic. In contrast, in males, arsenic exposure also induced similar reproductive and developmental toxicity; however, the adverse effects were mainly evident only in the medium and high dietary arsenic treatment groups. We also examined the sex-specific effects of dietary arsenic exposure on the expression of genes that regulate the hypothalamus–pituitary–gonadal–liver (HPG-L) axis in fish. The gene expression results indicated the downregulation of HPG-L axis genes in females irrespective of the arsenic treatment dose; however, the reduced expression of HPG-L axis genes in males was recorded only in the medium and high arsenic treatment groups. These observations suggest that chronic arsenic exposure in either females or males causes reproductive and developmental toxicity in zebrafish. However, these toxic effects are markedly higher in females than in males. Our results also suggest that arsenic can act as an endocrine disruptor and mediate reproductive and developmental toxicity by disrupting the HPG-L axis in zebrafish. Full article
(This article belongs to the Special Issue Endocrine-Disrupting Chemicals and Reproductive Toxicology)
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13 pages, 6032 KiB  
Article
Toxic Effects of Bisphenol AF Exposure on the Reproduction and Liver of Female Marine Medaka (Oryzias melastigma)
by Huichen Li, Jiahao Gao, Yue Liu, Yujia Ding, Yusong Guo, Zhongduo Wang, Zhongdian Dong and Ning Zhang
Animals 2024, 14(2), 222; https://doi.org/10.3390/ani14020222 - 10 Jan 2024
Cited by 6 | Viewed by 2449
Abstract
In recent years, bisphenol AF (BPAF) in aquatic environments has drawn attention to its ecological risks. This study aims to investigate the toxic effects of BPAF (188.33 μg/L) exposure for 30 days on female marine medaka (Oryzias melastigma). On the 10th [...] Read more.
In recent years, bisphenol AF (BPAF) in aquatic environments has drawn attention to its ecological risks. This study aims to investigate the toxic effects of BPAF (188.33 μg/L) exposure for 30 days on female marine medaka (Oryzias melastigma). On the 10th and 30th day of exposure, the toxicity was evaluated using histological analysis of the liver and ovaries and the transcription levels of genes related to the antioxidant system, immune system, and hypothalamic-pituitary-gonadal (HPG) axis. Findings revealed that (1) BPAF exposure caused vacuolation, karyopyknosis and karyolysis in the liver of marine medaka, and the toxic impact augmented with duration; (2) exposure to BPAF for 10 days facilitated the growth and maturation of primary ova, and this exposure had a comparatively inhibitory effect after 30 days; (3) exposure to BPAF resulted in a biphasic regulation of the transcriptional abundance of genes involved in antioxidant and inflammatory response (e.g., il-8, cat), with an initial up-regulation followed by down-regulation. Additionally, it disrupted the transcriptional pattern of HPG axis-related genes (e.g., 3βhsd, arα). In conclusion, 188.33 μg/L BPAF can alter the expression levels of functionally related genes, impair the structural integrity of marine organisms, and pose a threat to their overall health. Full article
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15 pages, 2908 KiB  
Article
Molecular Mechanisms of Tebuconazole Affecting the Social Behavior and Reproduction of Zebrafish
by Wei Yan, Guangyu Li, Qiqi Lu, Jianjun Hou, Meiqi Pan, Maomin Peng, Xitian Peng, Hui Wan, Xixia Liu and Qin Wu
Int. J. Environ. Res. Public Health 2023, 20(5), 3928; https://doi.org/10.3390/ijerph20053928 - 22 Feb 2023
Cited by 19 | Viewed by 2492
Abstract
The aim of this study was to explore the underlying mechanism of adverse effects caused by tebuconazole (TEB) on the reproduction of aquatic organisms In the present study, in order to explore the effects of TEB on reproduction, four-month-old zebrafish were exposed to [...] Read more.
The aim of this study was to explore the underlying mechanism of adverse effects caused by tebuconazole (TEB) on the reproduction of aquatic organisms In the present study, in order to explore the effects of TEB on reproduction, four-month-old zebrafish were exposed to TEB (0, DMSO, 0.4 mg/L, 0.8 mg/L, and 1.6 mg/L) for 21 days. After exposure, the accumulations of TEB in gonads were observed and the cumulative egg production was evidently decreased. The decline of fertilization rate in F1 embryos was also observed. Then the changes in sperm motility and histomorphology of gonads were discovered, evaluating that TEB had adverse effects on gonadal development. Additionally, we also found the alternations of social behavior, 17β-estradiol (E2) level, and testosterone (T) level. Furthermore, the expression levels of genes involved in the hypothalamic-pituitary-gonadal (HPG) axis and social behavior were remarkably altered. Taken together, it could be concluded that TEB affected the egg production and fertilization rate by interfering with gonadal development, sex hormone secretion, and social behavior, which were eventually attributed to the disruption of the expressions of genes associated with the HPG axis and social behavior. This study provides a new perspective to understanding the mechanism of TEB-induced reproductive toxicity. Full article
(This article belongs to the Special Issue Ecotoxicology and Health Risks of Emerging Organic Pollutants)
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21 pages, 4510 KiB  
Article
Endocrine Disruption of Propylparaben in the Male Mosquitofish (Gambusia affinis): Tissue Injuries and Abnormal Gene Expressions of Hypothalamic-Pituitary-Gonadal-Liver Axis
by Yun Ma, Yujing Li, Xiaohong Song, Tao Yang, Haiqin Wang, Yanpeng Liang, Liangliang Huang and Honghu Zeng
Int. J. Environ. Res. Public Health 2023, 20(4), 3557; https://doi.org/10.3390/ijerph20043557 - 17 Feb 2023
Cited by 4 | Viewed by 2660
Abstract
Propylparaben (PrP) is a widely used preservative that is constantly detected in aquatic environments and poses a potential threat to aquatic ecosystems. In the present work, adult male mosquitofish were acutely (4d) and chronically (32d) exposed to environmentally and humanly realistic concentrations of [...] Read more.
Propylparaben (PrP) is a widely used preservative that is constantly detected in aquatic environments and poses a potential threat to aquatic ecosystems. In the present work, adult male mosquitofish were acutely (4d) and chronically (32d) exposed to environmentally and humanly realistic concentrations of PrP (0, 0.15, 6.00 and 240 μg/L), aimed to investigate the toxic effects, endocrine disruption and possible mechanisms of PrP. Histological analysis showed time- and dose-dependent manners in the morphological injuries of brain, liver and testes. Histopathological alterations in the liver were found in 4d and severe damage was identified in 32d, including hepatic sinus dilatation, cytoplasmic vacuolation, cytolysis and nuclear aggregation. Tissue impairments in the brain and testes were detected in 32d; cell cavitation, cytomorphosis and blurred cell boundaries appeared in the brain, while the testes lesions contained spermatogenic cell lesion, decreased mature seminal vesicle, sperm cells gathering, seminiferous tubules disorder and dilated intercellular space. Furthermore, delayed spermatogenesis had occurred. The transcriptional changes of 19 genes along the hypothalamus–pituitary–gonadal–liver (HPGL) axis were investigated across the three organs. The disrupted expression of genes such as Ers, Ars, Vtgs, cyp19a, star, hsd3b, hsd17b3 and shh indicated the possible abnormal steroidogenesis, estrogenic or antiandrogen effects of PrP. Overall, the present results provided evidences for the toxigenicity and endocrine disruptive effects on the male mosquitofish of chronic PrP exposure, which highlights the need for more investigations of its potential health risks. Full article
(This article belongs to the Special Issue Toxicological and Health Effects of Environmental Contaminants)
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16 pages, 4850 KiB  
Article
Effects of 17α-Methyltestosterone on the Transcriptome and Sex Hormones in the Brain of Gobiocypris rarus
by Shaozhen Liu, Yue Chen, Tongyao Li, Liying Qiao, Qiong Yang, Weiya Rong, Qing Liu, Weiwei Wang, Jing Song, Xianzong Wang and Yu Liu
Int. J. Mol. Sci. 2023, 24(4), 3571; https://doi.org/10.3390/ijms24043571 - 10 Feb 2023
Cited by 16 | Viewed by 4139
Abstract
17α-Methyltestosterone (MT), a synthetic environmental endocrine disruptor with androgenic effects, has been shown to disrupt the reproductive system and inhibit germ cell maturation in Gobiocypris rarus. To further investigate the regulation of gonadal development by MT through the hypothalamic-pituitary-gonadal (HPG) axis, G. [...] Read more.
17α-Methyltestosterone (MT), a synthetic environmental endocrine disruptor with androgenic effects, has been shown to disrupt the reproductive system and inhibit germ cell maturation in Gobiocypris rarus. To further investigate the regulation of gonadal development by MT through the hypothalamic-pituitary-gonadal (HPG) axis, G. rarus were exposed to 0, 25, 50, and 100 ng/L of MT for 7, 14, and 21 days. We analyzed its biological indicators, gonadotropin-releasing hormone (GnRH), gonadotropins, reproduction-related gene expression, and brain tissue transcriptome profiles. We found a significant decrease in the gonadosomatic index (GSI) in G. rarus males exposed to MT for 21 days compared to the control group. GnRH, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) levels, as well as the expressions of the gnrh3, gnrhr1, gnrhr3, fshβ, and cyp19a1b genes, were significantly reduced in the brains of both male and female fish when exposed to 100 ng/L MT for 14 days compared to the controls. Therefore, we further constructed four RNA-seq libraries from 100 ng/L MT-treated groups of male and female fish, obtaining 2412 and 2509 DEGs in male and female brain tissue, respectively. Three common pathways were observed to be affected in both sexes after exposure to MT, namely, nicotinate and nicotinamide metabolism, focal adhesion, and cell adhesion molecules. Furthermore, we found that MT affected the PI3K/Akt/FoxO3a signaling pathway through the upregulation of foxo3 and ccnd2, and the downregulation of pik3c3 and ccnd1. Therefore, we hypothesize that MT interferes with the levels of gonadotropin-releasing hormone (GnRH, FSH, and LH) in G. rarus brains through the PI3K/Akt/FoxO3a signaling pathway, and affects the expression of key genes in the hormone production pathway (gnrh3, gnrhr1 and cyp19a1b) to interfere with the stability of the HPG axis, thus leading to abnormal gonadal development. This study provides a multidimensional perspective on the damaging effects of MT on fish and confirms that G. rarus is a suitable model animal for aquatic toxicology. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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11 pages, 2252 KiB  
Article
Effect of High-Fructose Diet-Induced Metabolic Syndrome on the Pituitary-Gonadal Axis in Male Rats
by Shih-Min Hsia, Yi-Fen Chiang, Hsin-Yuan Chen, Mohamed Ali, Paulus S. Wang and Kai-Lee Wang
Biomedicines 2022, 10(12), 3009; https://doi.org/10.3390/biomedicines10123009 - 23 Nov 2022
Cited by 6 | Viewed by 3488
Abstract
Plasma testosterone levels have been found to decrease in older insulin-resistant male patients. Both lower total testosterone levels and a higher incidence of metabolic syndrome have also been reported. The aim of this study was to investigate the effects of high-fructose diet-induced diabetes [...] Read more.
Plasma testosterone levels have been found to decrease in older insulin-resistant male patients. Both lower total testosterone levels and a higher incidence of metabolic syndrome have also been reported. The aim of this study was to investigate the effects of high-fructose diet-induced diabetes on both the testosterone release by Leydig cells and the activity of the hypothalamus–pituitary–gonadal (HPG) axis in male rats. Male rats were fed with either standard chow (control group) or a high-fructose diet (fructose-fed group) for 21 weeks. Hyperglycemia, hyperinsulinemia, and hypertension were observed in the fructose-fed group. Moreover, plasma testosterone and LH levels decreased in the fructose-fed group compared to the control group. Sperm motility was also reduced by 15% in the fructose-fed rats. In contrast, the basal release of testosterone from rat Leydig cells was not altered by fructose feeding. Moreover, in vitro studies showed that the testosterone release, in response to different stimulants, including forskolin (an adenylyl cyclase activator, 10−5 M), 8-Br-cAMP (a permeable analog of cAMP, 10−5 M), A23187 (a calcium ionophore, 10−5 M), or 25-hydroxy-cholesterol (water-soluble cholesterol, 10−5 M), did not significantly differ between the fructose-fed and control groups. Interestingly, the release of testosterone in response to human chorionic gonadotropin (hCG, 0.05 IU/mL) was enhanced by eightfold in the control group, but elevenfold in the fructose-fed group. LH receptor expression in rat Leydig cells was also increased. Moreover, LH secretion from the anterior pituitary was altered in the fructose diet-fed group. These results suggest that fructose diet-fed rats have lower plasma testosterone levels, which can lead to a higher sensitivity of hCG in Leydig cells. Full article
(This article belongs to the Special Issue Hormonal Regulation of Male Reproductive System)
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15 pages, 8101 KiB  
Article
Steroidogenic Effects of Salinity Change on the Hypothalamus–Pituitary–Gonad (HPG) Axis of Male Chinese Sea Bass (Lateolabrax maculatus)
by Zhenru Fang, Xujian Li, Yapeng Wang, Wei Lu, Juncheng Hou and Jie Cheng
Int. J. Mol. Sci. 2022, 23(18), 10905; https://doi.org/10.3390/ijms231810905 - 18 Sep 2022
Cited by 8 | Viewed by 2554
Abstract
As lower vertebrates, teleost species could be affected by dynamic aquatic environments and may respond to environmental changes through the hypothalamus–pituitary–gonad (HPG) axis to ensure their normal growth and sexual development. Chinese sea bass (Lateolabrax maculatus), euryhaline marine teleosts, have an [...] Read more.
As lower vertebrates, teleost species could be affected by dynamic aquatic environments and may respond to environmental changes through the hypothalamus–pituitary–gonad (HPG) axis to ensure their normal growth and sexual development. Chinese sea bass (Lateolabrax maculatus), euryhaline marine teleosts, have an extraordinary ability to deal with a wide range of salinity changes, whereas the salinity decrease during their sex-maturation season may interfere with the HPG axis and affect their steroid hormone metabolism, resulting in abnormal reproductive functioning. To this end, in this study, 40 HPG axis genes in the L. maculatus genome were systematically characterized and their copy numbers, phylogenies, gene structures, and expression patterns were investigated, revealing the conservation of the HPG axis among teleost lineages. In addition, freshwater acclimation was carried out with maturing male L. maculatus, and their serum cortisol and 11-ketotestosterone (11-KT) levels were both increased significantly after the salinity change, while their testes were found to be partially degraded. After salinity reduction, the expression of genes involved in cortisol and 11-KT synthesis (cyp17a, hsd3b1, cyp21a, cyp11c, hsd11b2, and hsd17b3) showed generally upregulated expression in the head kidneys and testes, respectively. Moreover, cyp11c and hsd11b2 were involved in the synthesis and metabolism of both cortisol and 11-KT, and after salinity change their putative interaction may contribute to steroid hormone homeostasis. Our results proved the effects of salinity change on the HPG axis and steroidogenic pathway in L. maculatus and revealed the gene interactions involved in the regulation of steroid hormone levels. The coordinated interaction of steroidogenic genes provides comprehensive insights into steroidogenic pathway regulation, as well as sexual development, in teleost species. Full article
(This article belongs to the Special Issue Endocrine Control of Fish Metabolism)
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13 pages, 4327 KiB  
Article
Life Cycle Exposure to Cyhalofop-Butyl Induced Reproductive Toxicity in Zebrafish
by Manman Duan, Xuanjun Guo, Xiangguang Chen, Mengyu Guo, Hao Xu, Lubo Hao, Chengju Wang and Yang Yang
Toxics 2022, 10(9), 495; https://doi.org/10.3390/toxics10090495 - 25 Aug 2022
Cited by 4 | Viewed by 2620
Abstract
Cyhalofop-butyl (CyB) is a herbicide widely used in paddy fields that may transfer to aquatic ecosystems and cause harm to aquatic organisms. In this study, zebrafish (Danio rerio) were exposed to CyB at environmental concentrations (0.1, 1 and 10 µg/L) throughout their adult [...] Read more.
Cyhalofop-butyl (CyB) is a herbicide widely used in paddy fields that may transfer to aquatic ecosystems and cause harm to aquatic organisms. In this study, zebrafish (Danio rerio) were exposed to CyB at environmental concentrations (0.1, 1 and 10 µg/L) throughout their adult life cycle, from embryo to sexual maturity. The effects of CyB on zebrafish growth and reproduction were studied. It was found that female spawning was inhibited, and adult male fertility decreased. In addition, we examined the expression of sex steroid hormones and genes related to the hypothalamus–pituitary–gonad–liver (HPGL) axis. After 150 days of exposure, the hormone balance in zebrafish was disturbed, and the concentrations of 17β-estradiol (E2) and vitellogenin (VTG) were decreased. Changes in sex hormone were regulated by the expression of genes related to the HPGL axis. These results confirmed that long-term exposure to CyB at environmental concentrations can damage the reproductive capacity of zebrafish by disrupting the transcription of genes related to the HPGL axis. Overall, these data may provide a new understanding of the reproductive toxicity of long-term exposure to CyB in zebrafish parents and offspring. Full article
(This article belongs to the Special Issue Exposure, Bioaccumulation and Toxicity of Pesticides)
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37 pages, 1077 KiB  
Review
Atrazine Exposure and Reproductive Dysfunction through the Hypothalamus-Pituitary-Gonadal (HPG) Axis
by Sara E. Wirbisky and Jennifer L. Freeman
Toxics 2015, 3(4), 414-450; https://doi.org/10.3390/toxics3040414 - 2 Nov 2015
Cited by 82 | Viewed by 13852
Abstract
Endocrine disrupting chemicals (EDC) are exogenous agents that alter endogenous hormone signaling pathways. These chemicals target the neuroendocrine system which is composed of organs throughout the body that work alongside the central nervous system to regulate biological processes. Of primary importance is the [...] Read more.
Endocrine disrupting chemicals (EDC) are exogenous agents that alter endogenous hormone signaling pathways. These chemicals target the neuroendocrine system which is composed of organs throughout the body that work alongside the central nervous system to regulate biological processes. Of primary importance is the hypothalamic-pituitary-gonadal (HPG) axis which is vital for maintaining proper reproductive function. Atrazine (2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine) is a pre-emergent herbicide used to prevent the growth of weeds on various crops. This herbicide is reported to widely contaminate potable water supplies everywhere it is applied. As such, the European Union banned the use of atrazine in 2004. Currently the United States Environmental Protection Agency regulates atrazine at 3 parts per billion (ppb; μg/L) in drinking water, while the World Health Organization recently changed their drinking water guideline to 100 ppb. Atrazine is implicated to be an EDC that alters reproductive dysfunction by targeting the HPG axis. However, questions remain as to the human health risks associated with atrazine exposure with studies reporting mixed results on the ability of atrazine to alter the HPG axis. In this review, the current findings for atrazine’s effects on the HPG axis are examined in mammalian, anuran, and fish models and in epidemiological studies. Full article
(This article belongs to the Collection Risk Assessment of Pesticide Exposure)
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