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Regional distribution of luteinizing hormone-releasing hormone (LH-RH) in rat brain determined by microdissection and radioimmunoassay.

We have re-examined the distribution of LH-RH in rat brain and in particular those rostral hypothalamic areas reported to contain perikarya by immunocytological techniques. Adult male rats were decapitated and 20 discrete brain structures micro-dissected. Tissue from single rats was homogenized in 50 microliter 0.1 N HCl and 40 microliters of this acid extract was radioimmunoassayed for LH-RH using the R-42 antibody. Low but detectable concentrations which ranged from0.2 to 1.0 pg/micrograms protein were found in the interstitial nucleus of the stria terminalis (ventral aspect), medial preoptic nucleus (dorsal and ventral aspects), lateral preoptic nucleus, anterior hypothalamic nucleus, paraventricular nucleus, ventromedial nucleus (medial and lateral aspects) and the dorsomedial nucleus. Slightly higher concentrations (1.0--4.0 pg/micrograms protein) were measured in a rostral, medial basal corridor made up of the suprachiasmatic preoptic nucleus, the suprachiasmatic nucleus and the retrochiasmatic area. LH-RH was not detected (i.e. less than 0.8 pg/area) in the bed nucleus of the diagonal band of Broca, caudate nucleus, lateral septal nucleus or the cingulate cortex.

Animals↗

Age-related changes of vasopressin content of microdissected areas of the rat brain.

Vasopressin contents of posterior pituitaries and intra- and extrahypothalamic nuclei of brains from 3-month and 24-month-old Long-Evans rats and from Brattleboro rats heterozygous (HEDI) and homozygous (HODI) for diabetes insipidus were measured by radioimmunoassay. The pituitary content of vasopressin was highest in young rats and was significantly lower in HEDI tissue. The peptide was not detected in HODI pituitaries. In the brain regions studied both aged and HEDI rats showed reduced vasopressin contents as compared to young animals. In light of evidence of memory deficiencies in both HEDI and HODI rats, the possibility arises that memory decrements associated with senescence may be related to altered vasopressin synthesis and/or secretion occurring with aging of these neuronal systems.

Aging↗

Differential responses in immunoreactive arginine-vasopressin content of microdissected brain regions during passive avoidance behavior.

Immunoreactive arginine-vasopressin (IR-AVP) was measured in various hypothalamic and extrahypothalamic nuclei of male Wistar rats immediately after the 24 h retention test of a passive avoidance response. IR-AVP concentrations in paraventricular, suprachiasmatic and lateral septal nuclei were significantly decreased in comparison with the non-shocked rats, while IR-AVP was increased in the central amygdala nucleus, subfornical organ and locus coeruleus. No significant differences in IR-AVP levels were found in the habenular and periventricular hypothalamic nucleus, organum vasculosum of lamina terminalis and medial and dorsal raphe nucleus.

Animals↗

gamma-Aminobutyric acid stimulates chloride permeability across microdissected Deiters' neuronal membrane.

Fluxes of 36Cl- across freshly prepared Deiters' neuronal membranes have been studied in a two-compartment microchamber simulating the extra- and the intracellular space. The rate of 36Cl- influx was enhanced by gamma-aminobutyric acid (GABA) (10(-4) M), the effect being reversed by picrotoxin (10(-4) M) and by bicuculline (10(-5) M). Diazepam (10(-8)-10(-7) M) did not potentiate the response to GABA and rather depressed it. However, a barbiturate site is most probably present in the GABA receptor complexes since pentobarbitone (10(-4) M) was able to stimulate 36Cl- permeability to the same extent as GABA itself.

Animals↗

Sex- and age-related differences in the activity of testosterone-metabolizing enzymes in microdissected nuclei of the zebra finch brain.

Many effects of testosterone (T) in the zebra finch (Taeniopygia guttata) can be mimicked by T-metabolites, mainly estradiol and 5 alpha-dihydrotestosterone. We have therefore studied the neuroanatomical distribution of testosterone-metabolizing enzymes by means of the Palkovits punch technique combined with radioenzyme assay in the brain of adult and young male and female zebra finches. The activity of these enzymes was studied by a one-point assay in 5 nuclei of the song system (X, MAN, HVc, RA, ICo), 2 nuclei of the visual system (ectostriatum, nucleus rotundus) and in limbic and hypothalamic areas. Very noticeable was the presence of a very high aromatase activity in the hippocampal and parahippocampal region and in the nucleus taeniae and the absence of this enzyme in ICo. We found a higher aromatase activity in female than male HVc and RA and a higher 5 alpha-reductase activity in MAN, HVc, RA and ICo of males compared to females. The 5 alpha-reductase was more active in the preoptic area of females. A few sex-related differences in the activity of the 5 beta-reductase were also observed (higher activity in females than in males for area X and RA, but difference in the opposite direction for the ectostriatum). The statistical significance of these differences depended, to some extent, on the statistical technique used to demonstrate them, with the sex differences in RA being by far the most robust ones. Many age-related metabolic differences were also detected but these do not have a clear interpretation since the Km of these enzymes also changes with age. Extremely low levels of 5 beta-reductase activity were found in the nuclei of the visual system in adult birds while this enzymatic activity was very high in young birds. The biological significance of this change with age remains obscure. Correlations are thus observed between the neuroanatomical distribution of T-metabolizing enzymes and of androgen and estrogen receptors with the important exception of ICo which has no aromatase but contains high concentrations of estrogen receptors. Testosterone-metabolizing enzymes are however also present in areas which are not known as steroid targets.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

Seasonal changes in luteinizing hormone-releasing hormone concentrations in microdissected brain regions of male rough-skinned newts (Taricha granulosa).

Luteinizing hormone-releasing hormone (LHRH) concentrations were measured in specific brain areas of male rough-skinned newts collected from a single population throughout the reproductive cycle. Plasma androgen and corticosterone (B) concentrations were also measured. Androgen concentrations were highest during the breeding season (winter) and lowest during the summer. In contrast, plasma B was lowest during the breeding season and highest in the summer. Concentrations of LHRH in the infundibulum (I) and rostral hypothalamus (RH) were positively correlated throughout the reproductive cycle; LHRH was always higher in the I than in the RH. Concentrations of LHRH in the ventral preoptic area (POA) fluctuated independently of concentrations of LHRH in the I and RH. However, LHRH concentrations decreased to undetectable levels in all three brain areas in March and April, which was before the end of the breeding season and before plasma androgen concentrations had decreased. An injection of LHRH into postbreeding males resulted in a significant increase in plasma androgen concentrations, indicating that the pituitary-gonad axis was still functional at the end of the breeding season. These results support the hypothesis that an abrupt decline in LHRH secretion is the initial endocrine event that signals the end of the breeding season in this amphibian.

Androgens↗