The hypothalamus plays a central role in the integrated control of feeding and energy homeostasis. We have identified two novel neuropeptides, both derived from the same precursor by proteolytic processing, that bind and activate two closely related (previously) orphan G protein-coupled receptors. These peptides, termed orexin-A and -B, have no significant structural similarities to known families of regulatory peptides. prepro-orexin mRNA and immunoreactive orexin-A are localized in neurons within and around the lateral and posterior hypothalamus in the adult rat brain. When administered centrally to rats, these peptides stimulate food consumption. prepro-orexin mRNA level is up-regulated upon fasting, suggesting a physiological role for the peptides as mediators in the central feedback mechanism that regulates feeding behavior.

No abstract

Light- and electron-microscopic immunocytochemistry (LM-ICC and EM-ICC) were used to visualize luteinizing hormone-releasing hormone (LHRH) in fibres associated with ventricular ependyma and tanycytes of the median eminence. LM-ICC suggests that LHRH fibers appear to enter the third ventricle. However, with EM-ICC, LHRH fibers are in fact found within ependymal canaliculi formed by adjacent ependymal cells. The canaliculi contain other myelinated and unmyelinated axons in addition to immunoreactive LHRH fibers. Thin slips of ependymal and tanycyte processes project into the canaliculi and enclose axons to varying degrees. At the median eminence many LHRH fibers bend sharply downwards from their ventricular course and travel with tanycytic processes towards their common destination - the perivascular space of the hypophysial-portal vascular system. Here, EM-ICC reveals that LHRH fibers closely contact basal processes of tanycytes. Lateral processes from tanycytes form glioplasmic sheaths which surround some individual LHRH fibers. A few LHRH terminals contact the perivascular space directly but more often are separated from the perivascular space by intervening glia. It is hypothesized that: (1) glia of this region responds to the physiological state of the animal and may determine the degree of LHRH secretion by varying the extent of glial investment of LHRH terminals; and (2) may play a role during development by providing direction and support for LHRH fibers similar to that described for radial and other glial cells.

Estrogen formation is catalyzed by the aromatase cytochrome P450 (P450AROM) enzyme. Aromatase activity has been detected in several regions in the rat brain. In the present study, we used peptide-generated polyclonal antibodies raised against a 20-amino acid synthetic fragment of the rat P450AROM protein (as deduced from the nucleic acid sequence of the rat P450AROM complementary DNA), to determine the location of this enzyme in rat brain sections. Immunoreactive antisera were titered by means of an enzyme-linked immunosorbent assay and purified by diethylaminoethyl-Affigel Blue chromatography. Specific immunoreactivity was confirmed by Western blot analysis using known aromatase-containing tissue (rat ovary homogenates and microsomal fractions). Evaluation of the distribution of P450AROM immunoreactivity in brain sections of male and female rats (30 and 60 days of age) was performed using the avidin biotin peroxidase immunocytochemical technique and light microscopy. P450AROM immunoreactivity appeared to be localized to neurons, and was present in brain regions and nuclei where enzymatic activity has been reported. For example, intense immunoreactivity was observed in the amygdaloid structures and supraoptic nucleus, whereas moderate to light immunoreactivity was evident in the paraventricular and arcuate nuclei and hippocampus. Surprisingly, neurons in the bed nucleus stria terminalis, medial basal hypothalamic, and preoptic areas displayed little aromatase immunoreactivity. However, P450AROM immunoreactivity was detected in specific brain regions not previously recognized to contain the enzyme (i.e. intense staining was seen in the reticular thalamic nucleus, olfactory tract and piriform cortex, as well as other brain structures). The pattern, distribution, and intensity of P450AROM immunoreactivity was similar regardless of sex or age. In this study, microsomal preparations derived from a new brain area (i.e. the reticular thalamic nucleus; Rt) displaying P450AROM immunoreactivity were observed to contain detectable levels of aromatase enzymatic activity, as determined by the 3H2O-release assay. The activity in the Rt was inhibited by a known aromatase inhibitor, 4-hydroxyandrostenedione. These results confirm histologically the localization of P450AROM to brain regions where aromatase enzymatic activity has been detected and extend the knowledge of its location to areas previously unknown as sites of aromatase activity, which may be involved in the modulation of neuroendocrine function and reproductive behavior.