Oestrogenic regulation of brain angiotensinogen.

Oestrogens are now recognized as playing a regulatory role on components of the systemic renin-angiotensin system, such as its precursor, angiotensinogen (AGT). In the brain, this role is poorly understood. The aim of this study was to investigate the influence of oestrogens on brain AGT of female rats at different stages of the oestrous cycle, in pregnancy and following ovariectomy with and without hormone replacement. AGT content of different brain regions was also studied in male rats treated with oestrogens. The brain was divided into five regions: cortex, cerebellum, brainstem, midbrain and thalamus/hypothalamus, and AGT was measured by direct radioimmunoassay using a highly specific AGT antibody. Cyclical fluctuations in AGT content were observed in all regions except the cerebellum over the course of the 4-day oestrous cycle, with peak concentrations at estrus and lowest concentrations at metestrus. Following ovariectomy, brain AGT was significantly decreased in the thalamic/hypothalamic region, an effect that was reversed by oestrogen-replacement. In pregnant rats, AGT contents were elevated in the brainstem region. Oestrogen treatment of male rats induced significant increases in AGT concentrations in all areas except the cortex. In summary, these results show that oestradiol has actions on brain AGT that are region-specific and dependent on the particular physiological and reproductive context. Moreover, the changes in AGT concentrations in the oestrous cycle suggest the involvement of other factors besides oestrogen. Finally, this study supports the view that the brain renin-angiotensin system has a broad role in oestrogen-modulated brain functions beyond those specific to the hypothalamic-pituitary-ovarian axis.

Growth hormone regulates AT-1a angiotensin receptors in astrocytes.

The hypothesis, based on previous in vivo data, that angiotensin AT1 receptors are regulated by GH or insulin-like growth factor I (IGF-I) has been investigated in this study using primary cultures of rat astrocytes as a model of AT1 receptor expression. At a dose of 1 ng/ml GH, there was an increase in AT1 density within 4 h and a maximum increase of 361 +/- 57% of the control value at 12 h. At 24 h, receptor density was still 176 +/- 23% that in the control. Astrocytes incubated with 1 ng/ml rat IGF-I for 24 h showed no change in AT1 receptor density. Reverse transcriptase-PCR was used to show that astrocytes express both the AT1a receptor subtype and, to a much lesser extent, the AT1b subtype. Treatment with 1 ng/ml recombinant bovine GH for 12 h increased the messenger RNA of the AT1a receptor by 170%, without affecting the AT1b receptor. Inhibition of protein synthesis by cycloheximide and of transcription by the adenosine analog dichlororibofuranosylbenzimidazole both prevented the increase in AT1 receptor density following GH treatment, indicating that the action of GH is transcriptional. In summary, we have shown that GH up-regulates, directly and not via IGF-I, angiotensin receptors of the AT1a subtype in astrocytes by a transcriptional mechanism. The long latency of the response and the dependency on transcription relegate the AT1a gene to the class of GH-regulated genes identified as delayed stable genes. This mechanism of AT1 activation may be one way in which GH activates the renin-angiotensin system and initiates consequential cardiovascular and angiogenic effects.

Short-term growth hormone (GH) treatment of GH-deficient adults increases body sodium and extracellular water, but not blood pressure.

Initiation of GH treatment in adults is frequently complicated by the development of symptomatic fluid retention. To investigate the mechanism and extent of fluid retention that occurs with dosages of GH used in the treatment of GH-deficient adults, we conducted a double blind study in which seven GH-deficient patients (aged 24-74 yr) each received in random order daily sc injections of placebo, a physiological dose of GH (0.04 U/kg, low dose), and a supraphysiological dose of GH (0.08 U/kg, high dose) for 7 days, separated by 21-day washout periods. On the seventh day, measurements were made of serum insulin-like growth factor I, body weight, exchangeable sodium, plasma volume, angiotensinogen, PRA, aldosterone, atrial natriuretic peptide (ANP), and mean 24-h ambulatory heart rate and blood pressure. GH significantly increased mean insulin-like growth factor I levels from 105 +/- 11 to 304 +/- 45 micrograms/L during low dose treatment (P = 0.006) and 400 +/- 76 micrograms/L during high dose treatment (P = 0.004). High dose GH caused a 1.2 +/- 0.3 kg increase in body weight (P = 0.01) and a 193 +/- 65 mmol increase in exchangeable sodium (P = 0.008). Low dose GH had a lesser effect, with no significant increase in body weight, but an increase in exchangeable sodium of 113 +/- 37 mmol (P = 0.02). Plasma volume was not significantly affected by GH treatment. Mean supine angiotensinogen levels were significantly higher during both GH treatments compared to placebo (low dose, P = 0.017; high dose, P = 0.028) as were mean supine PRA levels (low dose, P = 0.0002; high dose, P = 0.0025). Supine angiotensin II, aldosterone, and ANP levels were not significantly affected by GH treatment. There was no significant change from placebo in any of the sodium-regulating hormones in the erect posture. The mean 24-h heart rate was significantly higher during low dose (82 +/- 2 beats/min; P = 0.0001) and high dose (88 +/- 3 beats/min; P = 0.0001) GH treatment than during placebo (67 +/- 3 beats/min). However, no significant change in mean 24-h systolic or diastolic blood pressure was observed. In summary, acute GH administration using doses currently employed in treating adults causes a dose-related increase in body weight and body sodium, but no associated increase in blood pressure. We conclude that 1) sodium retention is a physiological effect of GH, but does not cause an acute rise in blood pressure; and 2) the mechanism of sodium and fluid retention is not primarily due to enhanced aldosterone secretion or inhibition of ANP release, but more likely to a direct renal tubular effect.

Posterior pituitary of the newborn marsupial possum, Trichosurus vulpecula.

The fetal anterior pituitary-adrenal axis is thought to be involved in the initiation of birth in both eutherian and marsupial mammals. Little is known about the structure and function of the posterior pituitary at birth in the marsupial. Immunocytochemistry, high pressure liquid chromatography, and radioimmunoassay were used to identify vasopressin and mesotocin in the posterior pituitary of a newborn marsupial, the brushtail possum, Trichosurus vulpecula. The concentrations of vasopressin and mesotocin in the head of the newborn possum were 0.34 and 0.28 ng, respectively. The concentration of vasopressin was always greater than that of mesotocin, and the amounts of neuropeptides present in the head increased as the possum developed.

Brain content and plasma concentrations of arginine vasopressin in an Australian marsupial, the brushtail possum Trichosurus vulpecula.

Arginine vasopressin (AVP) has been identified and quantified in the brain and plasma of the possum using a highly specific radioimmunoassay and high-performance liquid chromatography. Large amounts of AVP were found in the pituitary (16.3 +/- 0.56 micrograms/pituitary, n = 5) and hypothalamus (398 +/- 82.5 ng/hypothalamus), and significant amounts of AVP were also present in the cerebral cortex (26.8 +/- 11.5 ng/cortex). Plasma AVP concentrations were significantly lower (2.2 +/- 0.45 pg/ml, n = 10) during anesthesia compared to concentrations while conscious (4.5 +/- 1.19 pg/ml). Severe hemorrhage markedly increased plasma concentrations to 1091 +/- 225 pg/ml (n = 8). It was concluded that AVP is present in the possum brain, pituitary, and plasma, and that its secretion is stimulated by hypovolemia and inhibited by surgical stress.

Mesotocin and oxytocin in the brain and plasma of an Australian marsupial, the northern brown bandicoot, Isoodon macrourus.

1. Mesotocin (MT) and oxytocin (OT) were measured in the brain and plasma of bandicoots using reverse phase high performance liquid chromatography and specific radioimmunoassays. 2. MT and OT were found in the pituitary (1.25 +/- 0.10 micrograms/MT; 0.725 +/- 0.077 micrograms/OT) and hypothalamus (38.37 +/- 6.46 ng/MT; 19.1 +/- 4.61 ng/OT). Smaller amounts were present in the cerebral cortex. 3. Basal plasma concentrations ranged from 1.5 to 8.1 pg/ml for both peptides (N = 14) and were elevated by stress. 4. It was concluded that both MT and OT are secreted by the bandicoot brain and that stress stimulates secretion.

Immunocytochemical location of oxytocin and mesotocin within the hypothalamus of two Australian marsupials, the bandicoot Isoodon macrourus and the brushtail possum Trichosurus vulpecula.

The presence of oxytocin and mesotocin in the hypothalamus of two Australian marsupials, the bandicoot (Isoodon macrourus) and the brushtail possum (Trichosurus vulpecula), was examined by immunocytochemistry. Tissue was fixed in paraformaldehyde in phosphate buffer and immunoreactive cells were detected using highly specific rabbit antioxytocin and sheep anti-mesotocin as primary antisera. Immunoreactive oxytocin cells were demonstrated in the paraventricular and supraoptic nuclei of bandicoot and possum hypothalami, with greater density being observed in paraventricular nuclei. Immunoreactive mesotocin cells were also found in both hypothalamic nuclei of the possum but not of the bandicoot. The same cells appeared to stain for both peptides.

Regulation of rat brain angiotensin II (AII) receptors by intravenous AII and low dietary Na+.

Previous studies have shown the presence of specific AII receptors at several areas of the brain. The purpose of this study was to examine by radioreceptor assay the effect of intravenous AII infusion (5 or 25 ng/kg/min) and low dietary Na+ (less than 8 mmol/100 g) on AII receptors in five brain regions: the olfactory lobes (OLF), hypothalamus/thalamus/septum (HTS), midbrain (MID), cerebellum (CER) and medulla (MED). Scatchard analysis of binding data from control rats showed significant (P less than 0.01 ANOVA) differences between brain areas in both Ka (1.54 OLF, 1.87 HTS, 1.25 MID, 1.33 MED, 0.77 CER x 10(9) M-1) and Ro (321 OLF, 224 HTS, 203 MID, 145 MED, 41 CER fmol/g tissue). Following the i.v. infusion of AII for 4-7 days, marked changes were observed in the areas with a porous BBB, the HTS and MED. Both the Ka [3.20 (HTS) and 0.67 (MED) x 10(9) M-1] and Ro [116 (HTS) and 249 (MED) fmol/g tissue] changed. In addition, decreases in Ro were also observed in the OLF (241 fmol/g tissue) and CER (21 fmol/g tissue), areas which have not been considered as being accessible to blood-borne AII. A low Na+ diet for 21-30 days changed the Ka and Ro in all five regions but not in similar directions. Furthermore, with the exception of the OLF the direction of change was not similar to that caused by i.v. infusion of AII. It was concluded that AII receptor sites in the rat brain differ from each other in both receptor properties in their response to such regulatory factors as AII Na+ depletion.(ABSTRACT TRUNCATED AT 250 WORDS)