ABSTRACT
Methoxychlor (MXC), a commonly used pesticide, has been labeled as an endocrine disruptor. To evaluate the impact of neonatal exposure to MXC on female reproduction, female Sprague-Dawley rats were given subcutaneous injections on postnatal days 1, 3, and 5. The injections contained 1.0mg MXC, 2.0mg MXC, 10 µg 17ß-estradiol benzoate (positive control), or sesame oil (vehicle). The injections of MXC had no effect on anogenital distance or day of vaginal opening. Treatment with either 2.0mg MXC or estradiol significantly increased the total number of days with vaginal keratinization. Treatment with MXC had no effect on ability to exhibit a mating response as an adult female, although the high dose MXC (2.0) and the positive control (estradiol) animals demonstrated a decrease in degree of receptivity, a decrease in proceptive behavior and an increase in rejection behavior. These data suggest that higher doses of MXC given directly to pups during the neonatal period can act as an estrogen and alter aspects of the nervous system, impacting adult reproductive characteristics.
Subject(s)
Endocrine Disruptors/toxicity , Insecticides/toxicity , Methoxychlor/toxicity , Reproduction/drug effects , Sexual Behavior, Animal/drug effects , Age Factors , Animals , Animals, Newborn , Brain/drug effects , Brain/growth & development , Brain/metabolism , Endocrine Disruptors/administration & dosage , Estradiol/administration & dosage , Estradiol/analogs & derivatives , Estrogen Replacement Therapy , Female , Injections, Subcutaneous , Insecticides/administration & dosage , Methoxychlor/administration & dosage , Motor Activity/drug effects , Ovariectomy , Rats , Rats, Sprague-DawleyABSTRACT
Knockout (KO) mice lacking the orphan nuclear receptor steroidogenic factor 1 (SF-1) exhibit marked structural abnormalities of the ventromedial nucleus of the hypothalamus (VMH). In this study, we sought to determine the molecular mechanisms underlying the VMH abnormalities. To trace SF-1-expressing neurons, we used a SF-1/enhanced green fluorescent protein (eGFP) transgene. Although the total numbers of eGFP-positive cells in wild-type (WT) and SF-1 KO mice were indistinguishable, cells that normally localize precisely within the VMH were scattered more diffusely in adjacent regions in SF-1 KO mice. This abnormal distribution is likely due to the loss of SF-1 expression in VMH neurons rather than secondary effects of deficient steroidogenesis, as redistribution also was seen in mice with a CNS-specific KO of SF-1. Thus, the absence of SF-1 alters the distribution of cells that normally form the VMH within the mediobasal hypothalamus. Consistent with this model, the hypothalamic expression patterns of the transcription factors islet-1 and nkx2.1 also were displaced in SF-1 KO mice. Independent of gene expression, birthdate analyses further suggested that cells with earlier birthdates were affected more severely by the loss of SF-1 than were later born cells. We conclude that the absence of SF-1 causes major changes in cellular arrangement within and around the developing VMH that result from altered cell migration.
Subject(s)
Cell Movement/genetics , DNA-Binding Proteins/deficiency , Gene Expression Regulation, Developmental/genetics , Nervous System Malformations/pathology , Neurons/pathology , Transcription Factors/deficiency , Ventromedial Hypothalamic Nucleus/abnormalities , Adaptor Proteins, Signal Transducing/genetics , Adaptor Proteins, Signal Transducing/metabolism , Animals , Body Patterning/genetics , Cell Differentiation/genetics , Cell Division/genetics , DNA-Binding Proteins/biosynthesis , DNA-Binding Proteins/genetics , Female , Gonadal Steroid Hormones/biosynthesis , Homeodomain Proteins , Male , Mice , Mice, Knockout , Nervous System Malformations/genetics , Nervous System Malformations/metabolism , Neurons/metabolism , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Receptors, Cytoplasmic and Nuclear , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Stem Cells/cytology , Stem Cells/metabolism , Steroidogenic Factor 1 , Thyroid Nuclear Factor 1 , Transcription Factors/biosynthesis , Transcription Factors/genetics , Transcription Factors/metabolism , Transgenes/genetics , Ventromedial Hypothalamic Nucleus/metabolism , Ventromedial Hypothalamic Nucleus/pathologyABSTRACT
Estrogen receptor (ER) expression and regulation is vital to the correct functioning of the neuroendocrine brain. Islet-1 (Isl-1) is a LIM homeodomain-containing transcription factor that has been implicated in neuronal differentiation, is located in the hypothalamus, and can alter ER function in vitro. We have determined that Isl-1 is localized in several regions of the hypothalamus, including the ER rich areas of the ventromedial nucleus (VMH), the preoptic area, and the anterior hypothalamus. Using double-label immunocytochemistry, we examined the overlap between immunoreactive ERalpha and Isl-1 in these different hypothalamic brain regions. In the developing brain, almost 100% of VMH cells that contain immunoreactive ERalpha also contain Isl-1. However, in older animals, the percentage of double-label cells decreased below 70%. This change is due to a decrease in the number of cells containing Isl-1, because there was no difference in the number of ERalpha-containing cells. By contrast, in more anterior regions of the hypothalamus, cells containing both Isl-1 and ERalpha were less common, with the two populations adjacent to each other, rather than overlapping. These data suggest that, although Isl-1 and ERalpha can interact, they are not always found in the same cells and that regulation of ERalpha function is not under the same control in the VMH, preoptic area, and the anterior hypothalamus.
Subject(s)
Homeodomain Proteins/metabolism , Nerve Tissue Proteins , Preoptic Area/metabolism , Receptors, Estrogen/metabolism , Ventromedial Hypothalamic Nucleus/metabolism , Age Factors , Animals , Estrogen Receptor alpha , Female , Immunohistochemistry , LIM-Homeodomain Proteins , Male , Mice , Mice, Inbred C57BL , Pregnancy , Preoptic Area/growth & development , Transcription Factors , Ventromedial Hypothalamic Nucleus/growth & developmentABSTRACT
Gamma-aminobutyric acid (GABA) is a highly abundant neurotransmitter in the brain and the ligand for GABA(A), GABA(B), and GABA(C) receptors. Unlike GABA(A) and GABA(C) receptors, which are chloride channels, GABA(B) receptors are G-protein linked and alter cell-signaling pathways. Electrophysiological studies have found GABA(B) receptors in cultured embryonic hypothalamus, but the distribution of these receptors remains unknown. In the present study, we examined the expression of GABA(B) receptors in the ventromedial nucleus of the hypothalamus (VMH) during embryonic mouse development. GABA(B) receptors were present in the VMH at all ages examined, from embryonic day 13 to postnatal day 6. Using a brain slice preparation, we examined the effect of GABA(B) receptor activation on cell movement in the embryonic VMH as the nucleus forms in vitro. The GABA(B) receptor agonist baclofen decreased the rate of cell movement in a dose-dependent manner. Baclofen reduced cell movement by up to 56% compared with vehicle-treated controls. The percentage of cells moving per field and the angles of cell movement were not affected. With our previous findings of GABA(A) receptor activation, it is likely that GABA influences VMH development via multiple mechanisms.
