Heptachlor-induced epithelial to mesenchymal transition in HK-2 cells mediated via TGF-ß1/Smad signalling.

This study investigated the effect of heptachlor-induced oxidative stress (OS) on transforming growth factor (TGF)-ß1-mediated epithelial to mesenchymal transition (EMT) in human renal proximal tubular epithelial (HK-2) cells. Following treatment of HK-2 cells with an increasing concentration of heptachlor (0.01-10 µM) for 24 h, the intracellular reactive oxygen species and malondialdehyde level increased, whereas the glutathione-s-hydroxylase (GSH) level declined significantly in a dose-dependent manner. Pretreatment with N-acetyl cysteine attenuates the heptachlor-induced OS. In this study, we have shown that heptachlor-induced OS regulates the mRNA expression of TGF-ß1-mediated Smad signalling genes accompanied by increased nuclear localization of phosphorylated Smad-2 and phosphorylated Smad-3. Furthermore, the m-RNA and protein level of epithelial marker, that is, E-cadherin decreased while the mesenchymal marker, that is, α-smooth muscle actin increased in heptachlor exposed HK-2 cells. In conclusion, heptachlor-induced OS might be responsible for the activation of TGF-ß1/Smad signalling which ultimately leads to renal damage by means of EMT.

Combination of multiple neural crest migration assays to identify environmental toxicants from a proof-of-concept chemical library.

Many in vitro tests have been developed to screen for potential neurotoxicity. However, only few cell function-based tests have been used for comparative screening, and thus experience is scarce on how to confirm and evaluate screening hits. We addressed these questions for the neural crest cell migration test (cMINC). After an initial screen, a hit follow-up strategy was devised. A library of 75 compounds plus internal controls (NTP80-list), assembled by the National Toxicology Program of the USA (NTP) was used. It contained some known classes of (developmental) neurotoxic compounds. The primary screen yielded 23 confirmed hits, which comprised ten flame retardants, seven pesticides and six drug-like compounds. Comparison of concentration-response curves for migration and viability showed that all hits were specific. The extent to which migration was inhibited was 25-90%, and two organochlorine pesticides (DDT, heptachlor) were most efficient. In the second part of this study, (1) the cMINC assay was repeated under conditions that prevent proliferation; (2) a transwell migration assay was used as a different type of migration assay; (3) cells were traced to assess cell speed. Some toxicants had largely varying effects between assays, but each hit was confirmed in at least one additional test. This comparative study allows an estimate on how confidently the primary hits from a cell function-based screen can be considered as toxicants disturbing a key neurodevelopmental process. Testing of the NTP80-list in more assays will be highly interesting to assemble a test battery and to build prediction models for developmental toxicity.

Effects on the reproductive parameters of two generations of Rattus norvegicus offspring from dams exposed to heptachlor during gestation and lactation.

Heptachlor has been targeted for global elimination because of its toxicity and environmental persistence, in accordance with the Stockholm Convention on Persistent Organic Pollutants (POPs). However, there is no regulation of heptachlor use in Mexico, where relatively high levels have been found in maternal breast milk. The aim of this study was to determine the effects of heptachlor on the reproductive system of offspring of two consecutive generations of rats (F1 and F2) from dams orally administered heptachlor during midgestation and lactation. Female offspring were analyzed for different phenotypic, reproductive, and molecular parameters. In the F1 generation, heptachlor treatment induced decreased body weight at weaning, increased female anogenital distance, and delayed vaginal opening. In both generations, serum progesterone levels decreased and estradiol levels remained unchanged, while overexpression of the progesterone receptor was observed in uterine epithelial cells on estrus day. In the F2 generation, expression of the estrogen receptor α increased in the glandular epithelium. Finally, heptachlor treatment did not affect apoptosis in the uterine epithelial cells. Overall, the results indicate that heptachlor induced female reproductive alterations when administered to dams during the perinatal period. Accordingly, exposure to heptachlor may represent a risk for the reproductive health of humans. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 856-868, 2017.

Secretory function of ovarian cells and myometrial contractions in cow are affected by chlorinated insecticides (chlordane, heptachlor, mirex) in vitro.

The aim of the study was to investigate the effect of chlordane, heptachlor and mirex, on hormonal regulation of the force of myometrial contractions. Myometrial, endometrial, granulosa and luteal cells as well as strips of myometrium from non-pregnant cows were incubated with three insecticides at environmentally relevant doses (0.1, 1 or 10ng/ml). None of the insecticides affected the viability of studied cells. Chlordane stimulated, while heptachlor and mirex inhibited, secretion of testosterone and estradiol from granulosa cells as well as secretion of progesterone from luteal cells, respectively. Secretion of oxytocin (OT) from granulosa cells was increased after incubation with all studied insecticides. Only mirex stimulated OT secretion from luteal cells, while heptachlor inhibited this effect. None of them affected synthesis of OT in luteal cells and prostaglandins (PGF2 and PGE2) secretion from uterine cells, except PGE2 secretion from endometrial cells was decreased when the cells were incubated with 0.1ng/ml of chlordane. Basal and OT-stimulated myometrial contractions were increased by mirex and decreased by heptachlor. The data show that the insecticides altered secretory function of ovarian cells. Heptachlor and mirex affected also myometrial contractions in vitro, but uterine secretion of prostaglandins were not involved in the mechanism of that adverse effect of insecticides. The data indicate on potential of these insecticides to disturb fertilisation, blastocyst implantation or even the length of gestation.

Heptachlor induced nigral dopaminergic neuronal loss and Parkinsonism-like movement deficits in mice.

Epidemiological studies have suggested an association between pesticide exposure and Parkinson's disease. In this study, we examined the neurotoxicity of an organochlorine pesticide, heptachlor, in vitro and in vivo. In cultured SH-SY5Y cells, heptachlor induced mitochondria-mediated apoptosis. When injected into mice intraperitoneally on a subchronic schedule, heptachlor induced selective loss of dopaminergic neurons in the substantia nigra pars compacta. In addition, the heptachlor injection induced gliosis of microglia and astrocytes selectively in the ventral midbrain area. When the general locomotor activities were monitored by open field test, the heptachlor injection did not induce any gross motor dysfunction. However, the compound induced Parkinsonism-like movement deficits when assessed by a gait and a pole test. These results suggest that heptachlor can induce Parkinson's disease-related neurotoxicities in vivo.

Heptachlor induced nigral dopaminergic neuronal loss and Parkinsonism-like movement deficits in mice

Epidemiological studies have suggested an association between pesticide exposure and Parkinson's disease. In this study, we examined the neurotoxicity of an organochlorine pesticide, heptachlor, in vitro and in vivo. In cultured SH-SY5Y cells, heptachlor induced mitochondria-mediated apoptosis. When injected into mice intraperitoneally on a subchronic schedule, heptachlor induced selective loss of dopaminergic neurons in the substantia nigra pars compacta. In addition, the heptachlor injection induced gliosis of microglia and astrocytes selectively in the ventral midbrain area. When the general locomotor activities were monitored by open field test, the heptachlor injection did not induce any gross motor dysfunction. However, the compound induced Parkinsonism-like movement deficits when assessed by a gait and a pole test. These results suggest that heptachlor can induce Parkinson's disease-related neurotoxicities in vivo.

Degradation of the persistent organic pollutant [14C]heptachlor in Japanese field soils.

The fate of [(14)C]heptachlor in Saitama soil and the degradation of [(14)C]heptachlor in four Japanese field soils over 112 d after application were investigated. Heptachlor was degraded mainly to cis-heptachlor epoxide by a biotic process and to 1-hydroxychlordene by an abiotic process in the field soils. Volatilization of heptachlor and cis-heptachlor epoxide from the soil was observed over the experimental period. The amount of 1-hydroxychlordene produced in the soils appeared to be related to the soil water contents. Because heptachlor and heptachlor epoxides are predicted to volatilize to the atmosphere and to persist in soils, these compounds are thought to spread among Japanese environmental compartments even after a ban on their use.

Combined effects of organochlorine pesticides heptachlor and hexachlorobenzene on the promotion stage of hepatocarcinogenesis in rats.

We aimed to investigate the combined effect of organochlorine pesticides heptachlor (HEP) and hexachlorobenzene (HCB) by using a medium-term rat liver bioassay. Male F344 rats were initially administered diethylnitrosamine (DEN, 200mg/kgi.p.); after a 2-week non-dosing period, they were given diets containing HEP (5 or 25ppm), HCB (70 or 350ppm), or their mixtures (5 and 70ppm or 25 and 350ppm) for 6weeks. All rats were subjected to partial hepatectomy at week 3 and killed at week 8. We observed additive or synergistic effects of HEP and HCB in groups treated with mixtures of these pesticides. Number and area of preneoplastic foci positive for glutathione S-transferase placental form (GST-P) were consistently higher in these groups than the sum of individual values in the groups treated with HEP or HCB alone. Consistent with these findings, HEP and HCB had additive or synergistic effects on cell proliferation induction within the preneoplastic foci and cytochrome P450 (CYP) 2B1 and 3A1 induction, which may lead to more efficient metabolic activation of HEP and HCB. On the basis of these findings, we conclude that HEP and HCB have additive and synergistic effects on the development of GST-P-positive foci and that higher risks are associated with a combination of residual organochlorine pesticides in foods than with individual residual organochlorine pesticides.

Toxicity assessment and vitellogenin expression in zebrafish (Danio rerio) embryos and larvae acutely exposed to bisphenol A, endosulfan, heptachlor, methoxychlor and tetrabromobisphenol A.

Organochlorine pesticides and brominated flame retardants, such as tetrabromobisphenol A and polybrominated diphenyl ethers, pose an environmental hazard owing to their persistence, low solubility and estrogenic effects, and concerns have been raised regarding their effects on aquatic biota. In the present study, zebrafish embryos and larvae were used as a model to investigate the sublethal and lethal effects of three different organochlorine pesticides, namely methoxychlor, endosulfan and heptachlor, as well as the flame retardant tetrabromobisphenol A, and its precursor compound bisphenol A. Preliminary data for chemical exposure tests were obtained by determining the 96 h median effective concentration EC50 (hatching rate) and 96 h median lethal concentration LC50 . Quantitative polymerase chain reaction was used to investigate the gene expression levels of the biomarker vitellogenin (vtg1) after 96 h exposures to 10, 25, 50 and 75% of the 96 h EC50 value for embryos and 96 h LC50 value for larvae. The use of vtg1 mRNA induction in zebrafish embryos and larvae was found to be a sensitive biomarker of exposure to these organic compounds, and was helpful in elucidating their adverse effects and setting water quality guidelines.

Characteristic molecular signature for early detection and prediction of persistent organic pollutants in rat liver.

Persistent organic pollutants (POPs) are degradation-resistant anthropogenic chemicals that accumulate in the food chain and in adipose tissue, and are among the most hazardous compounds ever synthesized. However, their toxic mechanisms are still undefined. To investigate whether characteristic molecular signatures can discriminate individual POP and provide prediction markers for the early detection of POPs exposure in an animal model, we performed transcriptomic analysis of rat liver tissues after exposure to POPs. The six different POPs (toxaphene, hexachlorobenzene, chlordane, mirex, dieldrin, and heptachlor) were administered to 11-week-old male Sprague-Dawley rats, and after 48 h of exposure, RNAs were extracted from liver tissues and subjected to rat whole genome expression microarrays. Early during exposure, conventional toxicological analysis including changes in the body and organ weight, histopathological examination, and blood biochemical analysis did not reflect any toxicant stresses. However, unsupervised gene expression analysis of rat liver tissues revealed in a characteristic molecular signature for each toxicant, and supervised analysis identified 2708 outlier genes that discerned the POPs exposure group from the vehicle-treated control. Combination analysis of two different multiclassifications suggested 384 genes as early detection markers for predicting each POP exposure with 100% accuracy. The data from large-scale gene expression analysis of a different POP exposure in rat model suggest that characteristic expression profiles exist in liver hepatic cells and multiclassification of POP-specific molecular signatures can discriminate each toxicant at an early exposure time. The use of these molecular markers may be more widely implemented in combination with more traditional techniques for assessment and prediction of toxicity exposure to POPs from an environmental aspect.

Organochlorine pesticides induce epithelial to mesenchymal transition of human primary cultured hepatocytes.

Persistent organic pollutants (POPs) are a group of organic or chemicals that adversely affect human health and are persistent in the environment. These highly toxic compounds include industrial chemicals, pesticides such as organochlorines, and unwanted wastes such as dioxins. Although studies have described the general toxicity effects of organochlorine pesticides, the mechanisms underlying its potential carcinogenic effects in the liver are not well understood. In this study, we analyzed the effect of three organochlorine pesticides (dichlorodiphenyltrichloroethane, heptachlore and endosulfan) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the epithelial to mesenchymal transition (EMT) in primary cultured human hepatocytes. We found that these compounds modified the hepatocyte phenotype, inducing cell spread, formation of lamellipodia structures and reorganization of the actin cytoskeleton in stress fibers. These morphological alterations were accompanied by disruption of cell-cell junctions, E-cadherin repression and albumin down-regulation. Interestingly, these characteristic features of dedifferentiating hepatocytes were correlated with the gain of expression of various mesenchymal genes, including vimentin, fibronectin and its receptor ITGA5. These various results show that organochlorines and TCDD accelerate cultured human hepatocyte dedifferentiation and EMT processes. These events could account, at least in part, for the carcionogenic and/or fibrogenic activities of these POPs.

Organochlorine pesticides and polychlorinated biphenyls in sediments and fish from freshwater cultured fish ponds in different agricultural contexts in north-eastern France.

Organochlorine pesticides (HCB, HCH with α-, ß-, and γ isomers, heptachlor, cis-heptachlor epoxyde, trans-heptachlor epoxyde, endosulfan with α- and ß isomers, sulfate endosulfan, o,p'-DDT, p,p'-DDT, o,p'-DDE, p,p'-DDE, o,p'-DDD, p,p'-DDD, chlorothalonil, alachlor, aldrin, dieldrin, methoxychlor, oxychlordane, chlordane with α- and γ isomers, p,p'-dicofol and o,p'-dicofol) and indicators PCBs (IUPAC nos. 28, 52, 101, 118, 138, 153, and 180) were studied both in sediments and muscles of farmed fish species (Cyprinus carpio and Perca fluviatilis). Samples were collected from fish ponds located in the hydrographic basin of the Moselle River (Lorraine Region, France). OCPs and PCBs were present at low concentrations both in sediments and fish muscles. Concerning sediments, ∑DDTs revealed concentrations ranging from 0.2 to 2.30 ng g(-1) dw and ∑PCBs ranged from 0.3 to 3.5 ng g(-1) dw. Concerning fish muscles, the highest concentrations in OCPs were those of p,p'-DDE, with average concentrations of 0.57±0.44 ng g(-1) ww for carp and 0.58±0.29 ng g(-1) ww for perch. The contamination profiles proved to be different depending on the fish species. Indeed, HCH-isomers, HCB, and dieldrin were detected only for the carp and always at low concentrations. For example, the highest concentration of HCHs was observed for ß-HCH with a mean value of 0.64±0.15 ng g(-1) ww for carp. As for PCBs, the levels of ∑PCBs ranged from 0.3 to 6.4 ng g(-1) ww in carp muscles and from 0.90 to 5.60 ng g(-1) ww in perch muscles.

Phytotoxicity of heptachlor and endosulfan sulfate contaminants in soils to economic crops.

The intensive use of organochlorine in the past decades has resulted in contamination of soil worldwide. The phytotoxicity of two organochlorine pesticide, endosulfan sulfate and heptachlor, on the early growth stage of sweet corn (Zea mays), waxy corn (Zea mays) cowpea (Vigna sinensis), cucumber (Cucumis sativus) and water morning glory (Ipomoea aquatica) were studied. In the range of concentration found in Thai agricultural soil, 0.4-40 mg kg(-1) of each pesticide, did not affect the percentage of seed germination. Heptachlor seemed to affect the shoot and root length of test plants more than endosulfan sulfate. The combined effect of both pesticides to corn seedling growth was tested. There was no significant effect on combined treatment of both pesticides to corn growth. The 0.4-40 mg kg(-1) concentration of endosulfan sulfate and heptachlor did not produced significant effect on early growth of plants.

Genotoxicity of heptachlor and heptachlor epoxide in human TK6 lymphoblastoid cells.

The genotoxic potential of the organochlorine insecticides heptachlor (HC) and its metabolite heptachlor epoxide (HCE) has been evaluated in TK6 cells, a well-established human lymphoblastoid cell line. Genotoxicity has been determined by scoring the induction of DNA breaks in the comet assay and by measuring the frequency of micronuclei (MN) in binucleated cells. The results indicate that both compounds are able to induce significant increases in the percentage of DNA in the tail, the parameter used in the comet assay, with a direct dose-response relationship. Nevertheless, both compounds were unable to induce an increase in the frequency of MN. The comet assay measures primary DNA damage, while the induction of MN measures fixed damage. Thus, our results would suggest that the DNA damage induced by the two insecticides is not fixed as chromosome damage, which would be detectable by means of the MN assay (chromosome breaks and aneuploidy).

Exposure to heptachlor: evaluation of the effects on the larval and adult epidermis of Rana kl. esculenta.

Widely used in the past against termites and soil insects, the chlorinated insecticide heptachlor (H) is a toxic contaminant which represents a risk for both terrestrial and aquatic organisms. Like many organochlorine pesticides, heptachlor and heptachlor epoxide (HE), with oxidation products synthesized by many plant and animal species, degrade slowly since many of the derived compounds are persistent. This increases the status of heptachlor as a hazardous pollutant. In the present experimental study we exposed specimens of Rana kl. esculenta, from the tadpole stage through to their complete metamorphosis, to three different concentrations of heptachlor (4, 40 and 400 ppb). Mortality and HE bioaccumulation were evaluated on all the experimental groups. Since amphibian integument directly interacts with the environmental constituents (water, air and soil), we investigated the toxic effects on the ventral epidermis of both tadpole and adult samples by employing such histo-cytopathological biomarkers as ultrastructural morphology, certain enzyme activities (acid and alkaline phosphatases, AcPase, and AlkPase; succinic dehydrogenase, SDH; alpha-naphtyl butyrate esterase, ANBE; nitric oxide synthase/NADPH diaphorase, NOS/NADPHd). Also, the levels of reactive oxygen species (ROS) in the different conditions were evaluated. The results obtained were of ecological relevance, in particular as regards the effects of this environmental toxicant on the samples of tadpole epidermis. Severe morphological alterations were observed in the larval epidermal cells (apical and skein cells), whereas the cell epidermis (keratinocytes and mitochondria-rich cells) of the adult survivors showed changes in enzyme activities, particularly those involved in the protective response to xenobiotic injury. In general, morpho-histochemical studies, analysis of HE bioaccumulation and mortality showed a relation to the H doses employed.

Developmental heptachlor exposure increases susceptibility of dopamine neurons to N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)in a gender-specific manner.

Parkinson's disease (PD) is primarily thought of as a disease of aging. However, recent evidence points to the potential for exposure to xenobiotics during development to increase risk of PD. Here, we report that developmental exposure to the organochlorine pesticide heptachlor alters the dopamine system and increases neurotoxicity in an animal model of PD. Exposure of pregnant mice to heptachlor led to increased levels of the dopamine transporter (DAT) and vesicular monoamine transporter 2 (VMAT2) levels at both the protein and mRNA level in their offspring. Increased DAT and VMAT2 levels were accompanied by alterations of mRNA levels of nuclear transcription factors that control dopamine neuron development and regulate DAT and VMAT2 levels in adulthood. At 12 weeks of age, control and heptachlor-exposed offspring were administered a moderate dose (2 x 10mg/kg) of the parkinsonism-inducing agent MPTP. Greater neurotoxicity as evidenced by a greater loss of striatal dopamine and potentiation of increased levels of glial fibrillary acidic protein and alpha-synuclein was observed in heptachlor-exposed offspring. The neurotoxicity observed was greater in the male offspring than the female offspring, suggesting that males are more susceptible to the long-term effects of developmental heptachlor exposure. These data suggest that developmental heptachlor exposure causes long-term alterations of the dopamine system thereby rendering it more susceptible to dopaminergic damage in adulthood.

Pesticide use and chronic bronchitis among farmers in the Agricultural Health Study.

BACKGROUND: Farmers have increased risk for chronic bronchitis. Few investigators have considered pesticides. METHODS: We evaluated pesticides as risk factors for chronic bronchitis using the Agricultural Health Study enrollment data on lifetime pesticide use and history of doctor-diagnosed chronic bronchitis from 20,908 private pesticide applicators, primarily farmers. RESULTS: A total of 654 farmers (3%) reported chronic bronchitis diagnosed after age 19. After adjustment for correlated pesticides as well as confounders, 11 pesticides were significantly associated with chronic bronchitis. Heptachlor use had the highest odds ratio (OR=1.50, 95% Confidence Interval (CI)=1.19, 1.89). Increased prevalence for chronic bronchitis was also seen for individuals who had a history of a high pesticide exposure event (OR=1.85, 95% CI=1.51, 2.25) and for those who also applied pesticides in off-farm jobs (OR=1.40, 95% CI=1.04, 1.88). Co-morbid asthma and current farm activities did not explain these results. CONCLUSIONS: These results provide preliminary evidence that pesticide use may increase chronic bronchitis prevalence.

The toxic potential of aldrin and heptachlor on Danio rerio juveniles (Cypriniformes, Cyprinidae).

With the objective of evaluating the effects of organochlorine pesticides (aldrin and heptachlor) on the survival, growth and gill morphology of juvenile zebrafish (Danio rerio), four partial chronic toxicity bioassays were conducted (seven days' duration) with both compounds in a semi-static system with renewal every 24 h. The results did not show any effects on the fish's survival, but did on their growth and gill morphology. Aldrin was more toxic than heptachlor, since the chronic value (CV) for growth was nearly 8.7 times less, a result confirmed by analyzing the histology of the gills, in which the changes detected were considered more severe for the former substance. In general, they were found branchial lesions of first stage, in other words, cell proliferation between the secondary lamellae, hyperplasia, lifting of respiratory epithelial cells; fusion of several secondary lamellae and dilation of blood vessels. Aneurysms (alterations of second stage) they were verified in the exposed organisms to the aldrin, which also presented about 10% of reduction in the standard length and 30% of reduction in the total weight in relation to the exposed organisms to the control. For heptachlor these values were of 8% and 25%, respectively. The intensity of the gill lesions and growth of the fish did not depend on the pesticide concentration, suggesting different modes of action of the products.

The pesticide heptachlor affects steroid hormone secretion in isolated follicular and luteal cells of rat.

Heptachlor, a chlorinated hydrocarbon pesticide, suppresses the production of progesterone and estradiol in the female rat in vivo or in isolated ovaries in vitro. In this study the effect of heptachlor on steroid hormone production by isolated rat luteal and follicular cells, in the presence of two precursor hormones was investigated. Ovaries were isolated from anesthetized mature normocyclic virgin rats (3 to 4 months old), under sterile conditions. Corpora lutea and follicles were microscopically dissected out and separately enzymatically dispersed with collagenase at 37 degrees C. Viable cells collected after centrifugation were used at a concentration of approximately 2.5 x 10(5) cells/10 mL. Both luteal and follicular cell preparations were separately incubated overnight (15 h) at 37 degrees C in the presence of pregnenolone (P5) and androstenedione (A4) at a concentration of 6.0 nmol/L each, and heptachlor at either 0.12 microg/mL (low dose) or 1.20 microg/mL (high dose) (test cells) or in the absence of heptachlor (control cells). At the end of the incubations, progesterone and estradiol 17beta levels were analyzed in the incubation media. The results indicate that heptachlor significantly suppressed the production of both progesterone and estradiol in both cell types in a dose related manner even in the presence of A4 and P5 as precursor hormones (P<0.05).

Perinatal heptachlor exposure increases expression of presynaptic dopaminergic markers in mouse striatum.

Although banned in the 1970s, significant levels of the organochlorine pesticide heptachlor are still present in the environment raising concern over potential human exposure. In particular, organochlorine pesticides have been linked to an increased risk of Parkinson's disease. Studies from our laboratory and others have demonstrated that exposure of laboratory animals to heptachlor alters the levels and function of the dopamine transporter (DAT), an integral component of dopaminergic neurotransmission and a gateway for the dopaminergic neurotoxin MPTP. In this study, we examined the effects of developmental exposure to heptachlor on DAT, and other key components of the dopaminergic system, including the vesicular monoamine transporter 2 (VMAT2), tyrosine hydroxylase (TH), and aromatic amino acid decarboxylase (AADC). Female C57BL/6J mice received 0 or 3mg/kg heptachlor in peanut butter every 3 days for 2 weeks prior to breeding and throughout gestation and lactation until the offspring were weaned on postnatal day (PND) 21. On postnatal day 28, DAT, VMAT2, and TH levels were increased by 100, 70, and 30%, respectively, with no change in AADC levels or total dopamine levels. The ratio of DAT:VMAT2 was increased 29%. Since an increase in the DAT:VMAT2 ratio appears to predict susceptibility of brain regions to Parkinson's disease (PD) and results in increased toxicity of MPTP, these results suggest that alterations of the dopaminergic system by developmental heptachlor exposure may increase the susceptibility of dopamine neurons to toxic insult.