Recurrent insulin-induced hypoglycemia (RIIH) results in glucose counterregulatory dysfunction in men and male rodents. Intensified hypoglycemia in the latter coincides with diminished neuronal Fos expression in central metabolic regulatory structures, evidence that supports habituation of CNS-mediated compensatory motor outflow during re-exposure to this metabolic stress. In light of the evidence for counterregulatory resistance to precedent hypoglycemia in women, we utilized estradiol-treated ovariectomized (OVX) female rats to examine the hypothesis that this hormone regulates neural adaptability to recurring hypoglycemia. Groups of OVX rats were implanted with subcutaneous silastic capsules containing estradiol benzoate (E) or oil alone, and injected subcutaneously with one or four doses of the intermediate-acting insulin, Humulin NPH, one dose daily, or with diluent alone. Blood glucose levels were not altered by RIIH in E-implanted OVX animals, but were significantly decreased after four versus one insulin injection in the OVX+oil group. Mean numbers of Fos-immunoreactive (ir) neurons in the paraventricular nucleus hypothalamus (PVH), dorsomedial nucleus hypothalamus (DMH), and lateral hypothalamic area (LHA) were higher in both E- versus oil-implanted OVX rats injected with diluent only. Acute hypoglycemia significantly increased mean counts of Fos-ir-positive neurons in the PVH, DMH, and LHA, as well as the nucleus of the solitary tract (NTS) and area postrema (AP) in E- and oil-treated animals to an equivalent extent. OVX+E rats exhibited comparable numbers of Fos-positive neurons in the PVH, DMH, and LHA after one versus four insulin injections, whereas the numbers of labeled neurons in NTS and AP were increased or decreased, respectively, by RIIH. Oil-implanted OVX rats showed significantly diminished numbers of Fos-ir-positive neurons in each neural structure after repeated hypoglycemia. The present data demonstrate that estradiol sustains or enhances neuronal reactivity to recurring hypoglycemia in central metabolic structures, whereas hypoglycemic patterns of Fos expression in each site become habituated during RIIH in the absence of this steroid. The brain sites characterized here by estrogen-dependent maintenance of neuronal genomic reactivity to this substrate fuel imbalance may contain direct and/or indirect cellular targets for hormonal actions that prevent adaptation of CNS-controlled motor responses to this metabolic stress.

We examined indices of thyroid development in tadpoles from ammonium perchlorate (AP)-exposed sites. Bullfrog (Rana catesbeiana) tadpoles collected from a reference site exhibited normal developmental features, with many completing metamorphoses. In contrast, tadpoles collected from the AP contaminated site exhibited a 5-fold lower hindlimb/snout-vent length ratio than tadpoles from the reference site. The volume of the thyroid gland was 2.5-fold larger in the tadpoles from the reference site, presumably because they had progressed to late prometamorphosis and early metamorphic climax. Premetamorphic western chorus frog tadpoles (Pseudacris triseriata) inhabiting an ephemeral pond contaminated with AP exhibited gross morphological abnormalities of the thyroid including colloid depletion and follicle cell hypertrophy. We conclude that tadpoles exposed to AP-contaminated pond water early in larval life exhibit delayed development of thyroid-hormone sensitive structures. Additionally, there are abnormalities in the developing thyroid gland that seem to depend upon the window of AP exposure. The potential impact of thyroid disruption on development and reproduction in amphibian populations will be discussed.