Effects Of Atrazine Exposure Research Articles

Effects of Atrazine exposure on human bone marrow-derived mesenchymal stromal cells assessed by combinatorial assay matrix.

Mesenchymal Stromal/Stem cells (MSCs) are an essential component of the regenerative and immunoregulatory stem cell compartment of the human body and thus of major importance in human physiology. The MSCs elicit their beneficial properties through a multitude of complementary mechanisms, which makes it challenging to assess their phenotype and function in environmental toxicity screening. We here employed the novel combinatorial assays matrix approach/technology to profile the MSC response to the herbicide Atrazine, which is a common environmental xenobiotic, that is in widespread agricultural use in the US and other countries, but banned in the EU. Our here presented approach is representative for screening the impact of environmental xenobiotics and toxins on MSCs as an essential representative component of human physiology and well-being. We here employed the combinatorial assay matrix approach, including a panel of well standardized assays, such as flow cytometry, multiplex secretome analysis, and metabolic assays, to define the phenotype and functionality of human-donor-derived primary MSCs exposed to the representative xenobiotic Atrazine. This assay matrix approach is now also endorsed for characterization of cell therapies by leading regulatory agencies, such as FDA and EMA. Our results show that the exposure to Atrazine modulates the metabolic activity, size, and granularity of MSCs in a dose and time dependent manner. Intriguingly, Atrazine exposure leads to a broad modulation of the MSCs secretome (both upregulation and downmodulation of certain factors) with the identification of Interleukin-8 as the topmost upregulated representative secretory molecule. Interestingly, Atrazine attenuates IFNγ-induced upregulation of MHC-class-II, but not MHC-class-I, and early phosphorylation signals on MSCs. Furthermore, Atrazine exposure attenuates IFNγ responsive secretome of MSCs. Mechanistic knockdown analysis identified that the Atrazine-induced effector molecule Interleukin-8 affects only certain but not all the related angiogenic secretome of MSCs. The here described Combinatorial Assay Matrix Technology identified that Atrazine affects both the innate/resting and cytokine-induced/stimulated assay matrix functionality of human MSCs, as identified through the modulation of selective, but not all effector molecules, thus vouching for the great usefulness of this approach to study the impact of xenobiotics on this important human cellular subset involved in the regenerative healing responses in humans.

Effects of the Herbicide Atrazine on Crustacean Reproduction. Mini-Review.

Atrazine, one of the most intensively applied herbicides worldwide, is commonly found in several water bodies, affecting the associated fauna. Autochthon crustacean species have been relatively less studied, compared to vertebrate species, particularly concerning reproductive success. In this mini-review, we summarize the relevant information about the effects of atrazine exposure on the main reproductive aspects of crustaceans. One of these effects is related to the inhibition of ovarian growth. In this respect, a diminished vitellogenin content was found in the ovary of crabs exposed to atrazine during the entire period of ovarian growth, in correlation with a reduced oocyte size and a delay of ovarian maturation. Similar results were observed in crayfish. Atrazine was also able to affect the reproductive process, acting as an endocrine disruptor. In this sense, this herbicide was suspected to affect the secretion of some neurohormones involved in the gonadal growth, as well as to alter the circulating levels of steroid hormones which promote the synthesis of vitellogenin for ovarian growth. Moreover, atrazine induced sexual differentiation in juvenile crayfish toward a higher proportion of females, while it produced an increment of males in daphnids. Another aspect affected by this herbicide was the reduction of offspring production, as well as several embryonic abnormalities; genotoxic effects have been also reported in crayfish. Finally, some metabolic imbalances, such as reduction in energy reserves, have been observed in some species, together with oxidative stress and histopathological effects.

Development of scleral ossicles in Podocnemis expansa (Testudines: Podocnemididae) embryos exposed to atrazine

Understanding the effects of atrazine exposure on embryo development in oviparous animals may provide important data regarding the impacts of agrochemical use on wildlife and the ecosystem. This study set out to determine the effects of embryonic atrazine exposure on the development of osseous and cartilaginous components of scleral ossicles in Podocnemis expansa. Eggs were collected at the Environmental Protection Area Meandros do Rio Araguaia, Brazil, and artificially incubated in sand treated with solutions containing 2, 20 or 200 µg/L of atrazine. Sixty embryos were collected per treatment throughout the incubation period. Embryos were diaphanized with potassium hydroxide (KOH) and stained with Alizarin Red S and Alcian blue (bone and cartilage tissue respectively). Scleral ossicles were then counted and examined for skeletal abnormalities at different stages of embryonic development. Scleral ossicle counts were significantly reduced in P. expansa embryos treated with 200 μg/L atrazine solution. Rudimentary ossicles and gaps were also noted in embryos exposed to atrazine concentrations of 2 μg/L or 200 μg/L. Findings of this study emphasize the relevance of ecotoxicological investigations in determining the impacts of agrochemicals on native fauna.

Maternal exposure to atrazine induces the hippocampal cell apoptosis in mice offspring and impairs their learning and spatial memory

Background: The atrazine is widely used in the world as herbicide. It has been shown that atrazine can transmit from mother to their offspring via the placenta and the breast milk. Recent studies also confirmed that atrazine has ability to damage to the central nervous system. Since hippocampus is an important part of the brain that plays a key role in memory and learning, we evaluated the effects of atrazine exposure during pregnancy and lactation on the hippocampal cells and spatial memory in the male offspring of mice.Materials and methods: Twenty four pregnant Balb/C mice were divided into three groups of atrazine 10 mg/kg, atrazine 50 mg/kg and normal saline (n = 8). Atrazine and normal saline were given by gavage to the animals. Daily treatment with atrazine or normal saline began from gestational day (GD) 6 and continued until postnatal day (PD) 23. At the end of the treatments, brains of the male offspring were collected and histological studies were performed to detect apoptotic cells and dark neurons (DNs) in their hippocampus. Moreover, eight male offspring from each group were weaned and housed until adulthood (PD75). Then, learning and spatial memory were examined using Morris water maze (MWM) and passive avoidance (PA) tests.Results: The histological results of this study showed that apoptotic cells and DNs were significantly high in the offspring hippocampus in atrazine-exposed groups. Furthermore, the results of the behavioral tests indicated that learning and memory of offspring in the atrazine groups were significantly lower than in the normal saline group.Conclusions: Maternal exposure to atrazine during pregnancy and lactation periods increases the apoptotic cells and DNs in the hippocampus of the mice offspring and impairs their learning and spatial memory.

Atrazine exposure decreases the activity of DNMTs, global DNA methylation levels, and dnmt expression

Atrazine, a herbicide used on agricultural crops is widely applied in the Midwestern United States as well as other areas of the globe. Atrazine frequently contaminates potable water supplies and is a suspected endocrine disrupting chemical. Previous studies have reported morphological, hormonal, and molecular alterations due to developmental and adulthood atrazine exposure; however, studies examining epigenetic alterations are limited. In this study, the effects of atrazine exposure on DNA methyltransferase (DNMT) activity and kinetics were evaluated. Global DNA methylation levels and dnmt expression in zebrafish larvae exposed to 0, 3, or 30 parts per billion (ppb) atrazine throughout embryogenesis was then assessed. Results indicate that atrazine significantly decreased the activity of maintenance DNMTs and that the inhibition mechanism can be described using non-competitive Michaelis-Menten kinetics. Furthermore, results show that an embryonic atrazine exposure decreases global methylation levels and the expression of dnmt4 and dnmt5. These findings indicate that atrazine exposure can decrease the expression and activity of DNMTs, leading to decreased DNA methylation levels.

Effects of atrazine exposure on male reproductive performance in Drosophila melanogaster

Atrazine is a commonly utilized herbicide to control broadleaf weeds in the agricultural setting. It can, however, have negative effects on male reproductive performance in a variety of vertebrate species. Much less is known, however, about the effects of atrazine on invertebrates. In this study, we investigated the effects of several different concentrations of larval atrazine exposure on measures of reproductive performance in adult male Drosophila melanogaster. Atrazine exposure had significant effects on a male’s mating ability and the number of eggs his partner laid when he was successful at mating. Exposed males also sired a smaller proportion of the offspring under competitive conditions when they were the first male to mate to a doubly mated female. Atrazine exposure had no measurable effect on a male’s ability to prevent a mated female from mating to another male or on the proportion of offspring sired when the exposed males were the second male to mate. Exposure upregulated expression of one male reproductive gene, ovulin, but had no effect on expression of another, sex peptide. Exposed males produced and transferred more sex peptide protein to the female during mating but ovulin protein levels were not affected. In general, we observed non-monotonic responses such that the intermediate exposure levels showed the largest reduction in male reproductive performance. This study suggests that atrazine exposure affects male reproductive performance in insects and future studies should aim to understand the molecular mechanisms underlying the fitness effects of exposure.

Quantitative Assessment of Mammary Gland Development in Female Long Evans Rats Following In Utero Exposure to Atrazine

In this study, we quantified the effects of in utero exposure to the herbicide atrazine on subsequent mammary gland development. Atrazine was administered to pregnant female Long Evans rats from gestation days 13-19 at doses of 0, 6.5, 50, or 100 mg/kg/day. A pair-fed control group was yoked to the high-dose atrazine-treated group. Litter size was standardized to 10 pups on postnatal day (PND) 4. Whole mounts of the left fourth mammary gland and histologic sections of the right fourth gland were obtained from a subgroup of offspring on PND1, 21, 33, on day of vaginal opening (VO), or around PND65 at diestrus. A blinded, quantitative analysis of key morphological features in mammary gland whole mounts (ductal elongation, ductal network area, epithelial area, terminal end bud [TEB] incidence, and epithelial density) as well as epithelial proliferation within different parenchymal structures was conducted. There was no effect of atrazine exposure on any of the measures of mammary gland development at the maternal dose of 6.5 mg/kg/day. On PND1, ductal elongation was increased by approximately 20% (p < 0.05) in the female offspring born to dams exposed to 50 and 100 mg/kg/day atrazine, coincident with decreased epithelial proliferation in the 100 mg/kg/day group at this age. These differences were not present on PND21, or thereafter. An increased incidence of TEB in the mammary glands from females that were born to both the pair-fed and 50 mg/kg/day-treated dams at the time of VO indicated that this response was a specific result of maternal caloric restriction. Collectively, these data indicate that maternal atrazine exposure has no long-term effects on mammary gland development in female offspring beyond a transitory response to high doses at PND1.

Oxidative stress response and gene expression with atrazine exposure in adult female zebrafish ( Danio rerio)

Atrazine (ATZ) is the most common pesticide of freshwater ecosystems in the world. The effects of ATZ exposure on the induction of oxidative stress and the alteration of gene expression were studied in liver and ovary samples from female zebrafish ( Danio rerio). Antioxidant enzyme activities (SOD and CAT), in addition to the GSH and MDA content, in the liver altered significantly; the mRNA levels for the genes encoding these antioxidant proteins, such as Cu/Zn–Sod, Mn–Sod, Cat, and Gpx, were up-regulated significantly in the liver when zebrafish were exposed to various concentrations of ATZ for 14 d. However, the mRNA induction patterns were not in accordance with those of antioxidant enzyme changes in the liver and ovary. Moreover, the transcriptional expression of mitochondrial inner membrane genes related to ROS production, such as Ucp-2 and Bcl-2, were altered significantly in high ATZ treatment groups. However, the ATZ treatment did not induce any apparent changes in the transcriptional expression of CoxI, Ndi, or ATPo6, which are related to the mitochondrial respiratory chain and ATP synthesis. To our knowledge, this is the first report to study the transcriptional effects of ATZ stress on zebrafish. The information presented in this study will be helpful in fully understanding the mechanism of oxidative stress induced by ATZ in fish.

Intersex gonads in frogs: understanding the time course of natural development and role of endocrine disruptors

The paucity of data on sexual development of anuran amphibians has played an important role in the recent controversy over atrazine exposure. Although some studies have demonstrated the presence of abnormal gonads in control treatments, others have not, leading to varying interpretations of the effects of atrazine exposure on sexual development. However, the timing of development varies among anuran amphibians such that, at any snapshot in time, different species may exhibit different stages of sexual differentiation. We examined three species representing each of the differentiation rates (Bufo americanus=retarded rate; Hyla versicolor=basic rate; Rana sphenocephala=accelerated rate), to examine the natural time course of sexual development along with the influence of atrazine exposure. For each species, exposure to atrazine (1, 3, 10, 30 parts per billion), 17-beta-estradiol or control water occurred throughout larval life. Gonad histology was performed at 3-week intervals during the larval period or at a juvenile stage to examine the proportion of males, females, underdeveloped testes, testicular oocytes (TO; testes with 0-30% oocytes), and ovotestes (OVTs; testes with>30% oocytes). Our results illustrate that a phase of intersex gonads (TO or OVT) is normal during R. sphenocephala sexual development, a species representing the accelerated differentiation rate. Further, intersex gonads were found in juvenile stages of B. americanus and H. versicolor, representing retarded and basic rates, respectively, suggesting that a phase of intersex may be common regardless of differentiation rate. Moreover, these data highlight the importance of longitudinal studies rather than snapshots in time.

Low-level atrazine exposure decreases cell proliferation in human fibroblasts.

There is continuing concern that release of certain chemicals into the environment leads to human exposure to toxins, particularly through contaminated ground and surface waters. The triazine herbicide atrazine (2-chloro-4-ethylamine-6-isopropylamino-S-triazine) is routinely found as a contaminant in many surface and groundwaters. Little is known of the long-term effects of atrazine exposure on human cells. The objective of this research was to examine whether low-level exposure of atrazine to normal human fibroblast cells in culture had measurable deleterious effects. We show that low-level, short-term exposure of atrazine to normal fibroblasts results in decreased cell proliferation. In two separate assays to measure cell proliferation, either a 1.5-fold decrease in the cell number occurred (saturation density analysis) or a 2-fold decrease in cell proliferation was found (MTT analysis). Experiments to measure apoptosis in cells exposed to low levels of atrazine did not show DNA fragmentation or increased caspase activity. Thus, this analysis shows that short-term exposure to environmentally relevant, low levels of atrazine can be assessed by a decrease in cellular proliferation of normal human cells.

Atrazine induction of cytochrome P450 in Chironomus tentans larvae

Cytochrome P450-dependent aldrin epoxidation was characterized in third instar larvae of the aquatic midge, Chironomus tentans. Optimal in vitro assay conditions for the epoxidase were pH 7.6 and 31°C. Activity was linear up to 40 min of incubation time and 0.5 mg microsomal protein per incubation. The activity was concentrated in the microsomal fraction of whole body homogenates and was NADPH-dependent. The effect of atrazine exposure on aldrin epoxidase was measured to determine if this herbicide induces cytochrome P450-dependent activity. Comparisons of control and atrazine-exposed midges indicated increased epoxidase activity as a result of atrazine exposure, and a 45 kDa protein of increased intensity was observed after SDS-PAGE of microsomal protein. The molecular weight of this protein was similar in size to cytochrome P450 enzymes reported for other insects. Heme staining of SDS-PAGE gels and immunochemical studies using a Drosophila melanogaster anti-P450 polyclonal antiserum, further support the cytochrome P450 nature of this inducible 45 kDa protein.

Disorders of male rat reproductive tract under the influence of atrazine

The effects of atrazine exposure on testicular sperm number, epididymal sperm number and motility and alpha-glucosidase activity in the epididymis were studied in Fischer rats. Histological changes in the testicular tissue were followed by light and electron microscopy. Groups of adult animals were treated i.p. with 60 and 120 mg atrazine kg(-1) body wt. twice a week over 60 days. The results indicate a decrease in the body weight and relative weights of pituitary and ventral prostate vs control, measured on the last day of treatment in both treated groups. Testicular sperm number (expressed as number of sperm per 500 Sertoli cells) in atrazine-treated groups increased with the treatment time due to the reduced sperm motility. Therefore atrazine treatment provoked a significant decrease in sperm number and motility in epididymis, measured after the last day of treatment. alpha-Glucosidase activity in the epididymis, after the last day of treatment, showed a decrease in both treated groups vs control values. Histological analysis of testicular tissue from treated rats showed the cell disorganization and cell clusters together with spermatocytes. Electron microscopy presented differently vacuolated cytoplasm, collagen fibre was reduced, Leydig cells were of irregular shape with unequal form and cisternae of rough endoplasmic reticulum were accentuated and softly widened. In Sertoli cell cytoplasm, atrazine treatment provoked degenerative changes. According to the results obtained, it is evident that atrazine exerted morphological changes and a toxic effect on sperm and their motility.

Disorders of male rat reproductive tract under the influence of atrazine

The effects of atrazine exposure on testicular sperm number, epididymal sperm number and motility and α-glucosidase activity in the epididymis were studied in Fischer rats. Histological changes in the testicular tissue were followed by light and electron microscopy. Groups of adult animals were treated i.p. with 60 and 120 mg atrazine kg−1 body wt. twice a week over 60 days. The results indicate a decrease in the body weight and relative weights of pituitary and ventral prostate vs control, measured on the last day of treatment in both treated groups. Testicular sperm number (expressed as number of sperm per 500 Sertoli cells) in atrazine-treated groups increased with the treatment time due to the reduced sperm motility. Therefore atrazine treatment provoked a significant decrease in sperm number and motility in epididymis, measured after the last day of treatment. α-Glucosidase activity in the epididymis, after the last day of treatment, showed a decrease in both treated groups vs control values. Histological analysis of testicular tissue from treated rats showed the cell disorganization and cell clusters together with spermatocytes. Electron microscopy presented differently vacuolated cytoplasm, collagen fibre was reduced, Leydig cells were of irregular shape with unequal form and cisternae of rough endoplasmic reticulum were accentuated and softly widened. In Sertoli cell cytoplasm, atrazine treatment provoked degenerative changes. According to the results obtained, it is evident that atrazine exerted morphological changes and a toxic effect on sperm and their motility. Copyright © 2000 John Wiley & Sons, Ltd.