Immunohistochemical analyses of thyroid-specific enhancer-binding protein in the fetal and adult rat hypothalami and pituitary glands.

Thyroid-specific enhancer-binding protein (T/EBP), also known as NKX2.1 or TTF-1, regulates the expression of thyroid- and lung-specific genes. The t/ebp/Nkx2.1-null mutant mouse was stillborn but lacked the thyroid gland, pituitary gland, ventral region of the forebrain and normal lungs. These data demonstrated that T/EBP/NKX2.1 plays an important role not only in tissue-specific gene expressions in adults but also in genesis of these organs during development. Although the expression of t/ebp/Nkx2.1 in the brain has been reported, its function in the brain remains unknown. The present study was designed to determine the localization of T/EBP/NKX2.1 in the hypothalamus and pituitary gland of fetal and adult rats by immunohistochemistry as the first step toward understanding the function of T/EBP/NKX2.1 in the rat brain. In the fetal rat hypothalamus, T/EBP/NKX2.1 was localized widely in the ventral hypothalamic areas. In the adult rat brain, T/EBP/NKX2.1 was localized in the ventromedial hypothalamic nucleus, medial tuberal nucleus, arcuate nucleus and mammillary body. No T/EBP/NKX2.1 immunoreactivity was observed in the anterior or intermediate lobe of the pituitary gland in either fetal or adult rats. On the other hand, immunoreactive T/EBP/NKX2.1 was found in the posterior lobe of the pituitary gland. This paper presents results of detailed analyses of the distributions of T/EBP/NKX2.1 protein in the fetal and adult rat hypothalami and pituitary glands, and these results should provide important information for understanding the function of T/EBP/NKX2.1 in the brain.

Developmental increase in (3)H-muscimol binding to the gamma-aminobutyric acid(A) receptor in hypothalamic and limbic areas of the rat: why is the ventromedial nucleus of the hypothalamus an exception?

Using in vitro autoradiography to measure binding of the gamma-aminobutyric acid(A) (GABA(A)) receptor agonist, muscimol, we examined male and female rats on postnatal days 1, 5, 10, and 20. There were no sex differences in muscimol binding in any hypothalamic or limbic regions examined. However, all regions exhibited a developmental increase in the density of binding, except the ventromedial nucleus (VMN) of the hypothalamus. We have previously shown that the adult VMN is the only hypothalamic nucleus containing an abundance of the alpha2 subunit of the GABA(A) receptor and lack of the alpha1 subunit. We hypothesize that the lack of alpha1 may be causally related to the lack of increase in muscimol binding.

Ontogeny of region-specific sex differences in androgen receptor messenger ribonucleic acid expression in the rat forebrain.

Testosterone and its metabolites are the principal gonadal hormones responsible for sexual differentiation of the brain. However, the relative roles of the androgen receptor (AR) vs. the estrogen receptor in specific aspects of this process remain unclear due to the intracellular metabolism of testosterone to active androgenic and estrogenic compounds. In this study, we used an 35S-labeled riboprobe and in situ hybridization to analyze steady state, relative levels of AR messenger RNA (mRNA) expression in the developing bed nucleus of the stria terminalis, medial preoptic area, and lateral septum, as well as the ventromedial and arcuate nuclei of the hypothalamus. Each area was examined on embryonic day 20 and postnatal days 0, 4, 10, and 20 to produce a developmental profile of AR mRNA expression. AR mRNA hybridization was present on embryonic day 20 in all areas analyzed. In addition, AR mRNA expression increased throughout the perinatal period in all areas examined in both males and females. However, between postnatal days 4 and 10, sharp increases in AR mRNA expression in the principal portion of the bed nucleus of the stria terminalis and the medial preoptic area occurred in the male that were not paralleled in the female. Subsequently, males exhibited higher levels of AR mRNA than females in these areas by postnatal day 10. There was no sex difference in AR mRNA content in the lateral septum, ventromedial nucleus, or arcuate nucleus at any age. These results suggest that sex differences in AR mRNA expression during development may lead to an early sex difference in sensitivity to the potential masculinizing effects of androgen.

Prenatal ontogeny of growth hormone releasing hormone expression in rat hypothalamus.

The prenatal development of growth hormone-releasing hormone (GHRH) gene expression was studied in rat hypothalamus using hybridization histochemistry. Two populations of neurons expressing the GHRH gene were found and they had a different pattern of development. Neurons in the arcuate nucleus first expressed GHRH on E16 and 4 days later an adult-like pattern was found. Few neurons along the laterodorsal aspect of the ventromedial hypothalamic nucleus expressed GHRH on E17, with a subsequent increase of expression over the next days. On the day before birth, adult-like patterns were seen. These results demonstrate the differential regulation of GHRH gene expression in the rat hypothalamus.