RESUMO
Mesolimbic dopaminergic circuit is essential for food reward and motivational behaviors and can contribute to weight gain and obesity. Litter reduction is a classical model for studying the effects of neonatal overfeeding and overweight. Litters of Wistar rats were reduced to 4 pups/dam for small litter (SL) and 10 pups/dam for normal litter at postnatal day (PND) 4. Immediately after performing the feeding behavior tests, the animals were sacrificed in PND21 and PND90. The ventral tegmental area (VTA), Nucleus Accumbens Core (NAcC) and Shell (NAcSh) were isolated from frozen brain sections using the Palkovits micropunch technique. RNA and DNA were extracted from these areas, gene expression was measured by RT-qPCR and DNA methylation levels were measured by MSRM-qPCR technique. SL-PND21 animals presented increased expression levels of Tyrosine Hydroxylase and Dopamine Receptor D2 in VTA, decreased expression levels of dopamine active transporter (DAT) in VTA, and higher expression levels of DAT in NAcC. On the other hand, SL-PND90 animals showed decreased expression levels of Dopamine Receptor D1 and higher expression of DAT in NAcSh. These animals also evidenced impaired sensory-specific satiety. In addition, altered promoter methylation was observed at weaning, and remained in adulthood. This work demonstrates that neonatal overfeeding induces disruptions in the mesolimbic dopaminergic circuitry and causes alterations in feeding behavior from weaning to adulthood, suggesting that the neonatal period is critical for the normal development of dopaminergic circuit that impact on feeding behavior.
Assuntos
Metilação de DNA , Dopamina , Ratos , Animais , Dopamina/metabolismo , Ratos Wistar , Comportamento Alimentar , Núcleo Accumbens/metabolismoRESUMO
Environmental exposure to agrochemicals during early stages of development can induce subtle alterations that could permanently affect normal physiology. Previously, we reported that in ovo exposure to atrazine (ATZ) disrupts testicular histoarchitecture in postnatal caimans (Caiman latirostris). To assess whether such alterations are the result of disruption of gonadal developmental programming, this study aimed to evaluate the expression of histofunctional biomarkers (VASA, ER, PR, PCNA, and aromatase) and genes involved in gonadal development and differentiation (amh, sox-9, sf-1 and cyp19-a1) in the gonads of male and female caiman embryos and to assess the effect of ATZ exposure on these biomarkers and genes in the gonads of male embryos. Our results suggest that amh, aromatase and sox-9 play a role in sex determination and gonadal differentiation. In male caiman embryos, ATZ exposure increased aromatase expression and altered the temporal expression pattern of amh and sox-9 evidencing an ATZ-induced disruption of gonadal developmental programming. Since the effects of ATZ are consistent across all vertebrate classes, the ATZ-mediated disruptive effects here observed could be present in other vertebrate species.
Assuntos
Jacarés e Crocodilos , Atrazina , Animais , Feminino , Masculino , Atrazina/metabolismo , Aromatase/metabolismo , Gônadas , TestículoRESUMO
Agrochemicals or pesticides are compounds widely used to prevent, destroy or mitigate pests such as insects, rodents, herbs and weeds. However, most of them also act as environmental estrogens, anti-estrogens and/or antiandrogenic chemicals. In addition, both herbicides (such as glyphosate and paraquat) and insecticides (such as pyrethroids, organophosphates, neonicotinoids and rotenone) have been shown to exert significant adverse effects on hippocampal neurogenesis. These effects are particularly important because neurogenesis dysregulation could be associated with cognitive decline and neuropathologies such as Alzheimer's disease. This review focuses on the most commonly used agrochemicals in Argentina and their effects on the hippocampal neurogenesis of mammals. It also discusses the disruption of hormone synthesis and action as a possible mechanism through which these chemical compounds could alter the brain functions. Finally, we propose some lines of research to study the potential endocrine mechanisms involved in the effects of agrochemicals on human health and biodiversity.
Assuntos
Agroquímicos/toxicidade , Neurogênese/efeitos dos fármacos , Animais , Sistema Endócrino/efeitos dos fármacos , Herbicidas/toxicidade , Humanos , Inseticidas/toxicidade , Praguicidas/toxicidadeRESUMO
Recently, we have described the ontogeny of histofunctional differentiation changes in the oviduct of Caiman latirostris. The expression of estrogen receptor alpha and progesterone receptor shows that the caiman oviduct could be a target of the action of xenoestrogens such as the widely environmentally present Bisphenol A (BPA), early in life. The aims of this study were: to complement oviduct characterization by establishing the ontogenetic changes in androgen receptor (AR) expression and assessing the effects of early postnatal exposure to 17-ß-estradiol (E2) or BPA on the histofunctional features of the oviduct. AR was expressed in all the stages studied. The spatial pattern of AR immunostaining changed from neonatal to juvenile caimans. In the luminal epithelium, changes were at the subcellular level, from cytoplasmic to nuclear. In the subepithelium, although both cytoplasmic and nuclear AR expression was observed, changes were mainly at tissue level, from the subepithelial compartment to the outer muscular layer. The oviduct was highly sensitive to E2 and BPA at the early postnatal developmental stage. E2- and BPA-exposed caimans showed increased luminal epithelial height and higher proliferative activity. Changes in histomorphological features (measured by a scoring system), steroid hormone receptors, collagen remodeling and muscle-associated proteins suggest a precocious oviduct histofunctional differentiation in E2- and BPA-exposed caimans. The modification of the temporal pattern of oviductal biomarkers suggests that organizational changes could impair C. latirostris reproductive health later in life. The alterations in the caiman female reproductive tract exposed to BPA highlight the importance of preserving aquatic environments from plastic pollution.
Assuntos
Jacarés e Crocodilos/metabolismo , Compostos Benzidrílicos/toxicidade , Genitália Feminina/metabolismo , Genitália Feminina/patologia , Fenóis/toxicidade , Animais , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Estradiol/farmacologia , Feminino , Proteínas Musculares/metabolismo , Oviductos/efeitos dos fármacos , Oviductos/metabolismo , Receptores Androgênicos/metabolismo , Receptores de Progesterona/metabolismo , Fatores de TempoRESUMO
Removing dietary phyto-oestrogens in adult male rats causes obesity and diabetes. As whey proteins have been reported to reduce food intake and improve glucose homoeostasis, we investigated whether they could attenuate susceptibility to obesity and diabetes due to phyto-oestrogen deprivation. To this end, thirty male Wistar rats were fed a high-phyto-oestrogen (HP) or a phyto-oestrogen-free (PF) diet for 10 weeks; six rats from each group were killed. The remaining HP animals (six animals) continued receiving the HP diet for 6 weeks. The remaining PF rats (twelve rats) were divided in two groups: one was given the PF diet and the other a variation of the PF diet plus whey protein (PF-W). Body weight, food intake and adipose tissue weights were recorded. Hypothalamic mRNA expressions of orexigenic (neuropeptide Y, agouti-related protein (AgRP)) and anorexigenic (pro-opiomelanocortin (POMC), cocaine-amphetamine-related transcript (CART)) neuropeptides were quantified by real-time PCR. Serum glucose, insulin and total thyroxine (T4), thyroid-stimulating hormone, testosterone and oestradiol were assessed. After 10 weeks of PF diet, increased body weight, adiposity and energy intake, with up-regulation of AgRP and down-regulation of POMC', were observed. Longer treatment exacerbated these results, increased total T4 levels, reduced oestradiol levels and impaired glucose homoeostasis. PF-W reduced energy intake and increased POMC expression; however, body weight and adiposity remained unchanged. PF-W could not prevent the hormonal changes or the high circulating glucose levels induced by phyto-oestrogen deprivation, but reduced fasting insulin. These data demonstrate that, although 6 weeks of whey administration could not prevent obesity in phyto-oestrogen-deprived rats, the reduction in energy intake and circulating insulin could be beneficial with longer treatments.
Assuntos
Ingestão de Energia/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Hipotálamo/metabolismo , Obesidade , Proteínas do Soro do Leite/farmacologia , Ração Animal/análise , Animais , Glicemia , Dieta , Suplementos Nutricionais , Masculino , Fitoestrógenos/administração & dosagem , Fitoestrógenos/farmacologia , RNA/genética , RNA/metabolismo , Ratos , Reação em Cadeia da Polimerase em Tempo RealRESUMO
The increased use of agrochemical pesticides, such as atrazine (ATZ) and endosulfan (END), may have a significant impact on ecosystem health and biodiversity. The aim of this study was to investigate the consequences of in ovum exposure to ATZ and END on Caiman latirostris gonadal histo-functional features. Caiman eggs were collected from environmentally pristine areas and incubated in controlled conditions at male producing temperature (33 degrees C). At stage 20 of embryonic development, the sensitive stage for gonadal sex determination, eggs were exposed to one dose of either END or ATZ. Gonadal histo-morphology was examined in caiman hatchlings and serum levels of testosterone were measured. Regardless of treatment condition, all eggs incubated at 33 degrees C resulted in male hatchlings. Tortuous seminiferous tubules with increased perimeter, disrupted distribution of peritubular myoid cells (desmin positive), and emptied tubular lumens characterized the testes of pesticide-exposed caiman. An imbalance between proliferative activity and cell death was observed in the testes of caiman exposed to the higher doses of END, mainly due to a high frequency of apoptosis in intratubular cells. This altered cell turnover was associated with decreased testosterone levels. Prenatal exposure to only one dose of END and ATZ disrupted neonatal male gonadal histo-functional features. Alterations described here could have detrimental effects on the sexual maturation of the caiman and, ultimately, on the success of male caiman reproduction.
Assuntos
Jacarés e Crocodilos/embriologia , Exposição Ambiental/efeitos adversos , Praguicidas/farmacologia , Testículo/anormalidades , Testículo/efeitos dos fármacos , Testosterona/sangue , Zigoto/efeitos dos fármacos , Actinas/metabolismo , Jacarés e Crocodilos/crescimento & desenvolvimento , Animais , Animais Recém-Nascidos , Apoptose , Atrazina/farmacologia , Proliferação de Células , Desmina/metabolismo , Endossulfano/farmacologia , Masculino , Túbulos Seminíferos/metabolismo , Túbulos Seminíferos/patologia , Testículo/metabolismo , Testículo/patologia , Zigoto/crescimento & desenvolvimentoRESUMO
Human and wildlife are exposed at critical periods of development to endocrine disruptor chemicals (EDC) that may be responsible for reproductive disorders. To test the hypothesis that in ovum exposure to EDC at a critical period for gonadal organogenesis alters post-hatching folliculogenesis and steroidogenesis in Caiman latirostris, we studied the impact of in ovum exposure to 17 beta-estradiol (E2), bisphenol A (BPA), endosulfan (END) and atrazine (ATZ) on gonadal differentiation, follicular dynamics and circulating levels of steroid hormones in neonatal and juvenile caiman. Since C. latirostris is a species with temperature dependent sex determination, eggs were incubated at male (33 degrees C) or female (30 degrees C) producing temperatures and the effect of EDC was evaluated. Neonatal ovaries exhibited germ cells mainly located in clusters evidencing proliferative activity and type I to III follicles. Juvenile ovaries exhibited germ cells and advanced stages of pre-vitellogenic follicles. Prenatal exposure to the highest doses of E2 (1.4 ppm) or BPA (140 ppm) overrode male temperature effect on sex determination. Neonatal females produced by sex reversion lacked type III follicles, while females prenatally exposed to the lowest doses of E2 (0.014 ppm) and BPA (1.4 ppm) or ATZ (0.2 ppm) showed an increase in type III follicles. Juvenile caiman prenatally exposed to E2 or BPA showed an augmented incidence of multioocyte follicles. Neonatal female caiman exposed in ovum to E2 or BPA had higher estrogen serum levels whereas exposure to E2, BPA, ATZ and END decreased T levels. Present data demonstrates that exposure to EDC during gonadal organogenesis alters follicular dynamics and steroid levels later in life. These effects might have an impact on caiman fertility.
Assuntos
Jacarés e Crocodilos/metabolismo , Disruptores Endócrinos/farmacologia , Folículo Ovariano/metabolismo , Esteroides/metabolismo , Animais , Animais Recém-Nascidos , Atrazina/farmacologia , Compostos Benzidrílicos , Proliferação de Células/efeitos dos fármacos , Endossulfano/farmacologia , Estradiol/farmacologia , Feminino , Fertilidade/efeitos dos fármacos , Herbicidas/farmacologia , Inseticidas/farmacologia , Oócitos/efeitos dos fármacos , Oócitos/crescimento & desenvolvimento , Oócitos/ultraestrutura , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/crescimento & desenvolvimento , Fenóis/farmacologiaRESUMO
Organochlorine compounds (OCCs), such as pesticides and polychlorinated biphenyls (PCBs), are persistent lipophilic chemicals identified as endocrine disruptors, mainly with estrogen-like effects. Northeastern Argentina, near the Paraná River, is characterized by intensive farming and agricultural activities and industrial development, and is therefore prone to high incidences of environmental and dietary exposure to OCCs. Hence, we conducted a study to (1) estimate the organochlorine residues present in mammary fat tissue in a population of women from this littoral region and (2) identify potential sources of exposure to OCCs. Our subjects were 76 women (residing in and around Santa Fe city and not occupationally exposed to organochlorines) who underwent excision biopsy of a breast lesion or had plastic surgery. Both frequency of occurrence and levels of organochlorine residues were high in breast adipose tissue of all the participants. The organochlorine residues most frequently found were p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) in all the subjects analyzed, hexachlorobenzene (HCB) in 86.8%, and beta-hexachlorocyclohexane (beta-HCH) in 75.0%. The incidence of PCB congeners was very low. p,p'-DDE and beta-HCH residues reached the highest levels, 4794 and 1780 ppb, respectively. The diet was a relevant source of exposure, consumption of animal fat and freshwater fish playing a significant role. Bioaccumulation was evidenced by the significant positive association between organochlorine levels and body mass index (p = 0.0003) and the age of the patient (p = 0.0002). The frequency and levels of OCCs found in our study population raise concerns regarding Argentinean exposure to these endocrine disruptors.
Assuntos
Tecido Adiposo/metabolismo , Poluentes Atmosféricos/farmacocinética , Disruptores Endócrinos/farmacocinética , Exposição Ambiental/efeitos adversos , Hidrocarbonetos Clorados/farmacocinética , Poluentes Atmosféricos/intoxicação , Argentina , Mama/metabolismo , Disruptores Endócrinos/intoxicação , Feminino , Humanos , Hidrocarbonetos Clorados/intoxicação , Pessoa de Meia-Idade , Estatísticas não Paramétricas , População UrbanaRESUMO
Environmental pollution with endocrine disrupting compounds (EDCs) has adverse effects on the ecosystem's health. Caiman latirostris are widely distributed in South American aquatic ecosystems. Caimans have physiological and ecological characteristics that make them particularly vulnerable to EDCs exposure and suitable candidate as a sentinel species. Vitellogenin (Vtg) is a yolk pre-cursor protein synthesized by the liver of non-mammalian vertebrates and induced in response to estrogen. Purified plasma Vtg from caimans injected with estradiol-17beta (E2) was used to generate a polyclonal anti-body. Anti-body specificity was assessed using Western blot. The antiserum was also effective in detecting turtle Vtg, exhibiting high cross-reactivity with Vtg from Phrynops hilarii and Trachemys scripta dorbigni. We developed a specific and highly sensitive ELISA for caiman Vtg. This method has a detection limit of 0.1 ng/ml of plasma. The ELISA did not detect Vtg in plasma of non-induced male caimans. Induction of Vtg in male caimans was evaluated in response to one or two (7 days apart) doses of E2. Due to its high sensitivity, ELISA allows to measure the small increases in plasma Vtg after exposure to exogenous estrogen. A priming effect was observed following the second E2 dose, with a tenfold increase in circulating Vtg. Hepatic synthesis was confirmed by immunohistochemistry. The results presented herein suggest that detection of plasma Vtg in male caimans might become a valuable tool in biomonitoring xenoestrogen exposure in a polluted environment.