Anorectic effect of dehydroepiandrosterone combined with dexfenfluramine or thionisoxetine.

Free feeding rats given supplementary 1 h access per day to a palatable dessert test meal were tested for the anorectic effect of dehydroepiandrosterone alone or in combination with either the serotonin releasing agent dexfenfluramine or the norepinephrine uptake inhibitor thionisoxetine (LY 368975). Isobolographic analysis showed that the effect of dehydroepiandrosterone combined with either dexfenfluramine or thionisoxetine was within the range predicted for additivity of action.

Expression of Fos immunoreactivity in rat brain during dehydration: effect of duration and timing of water deprivation.

Water deprivation induces expression of the immediate early gene c-fos in specific brain regions, most likely as a result of the activation of cells that are responsive to changes in osmolality and/or blood volume. We hypothesized that the magnitude of c-fos expression would be a function of both the duration of water deprivation and the time of day at which the deprivation started. This study was designed to examine the pattern of Fos-like immunoreactivity (FLI) following water deprivation in rats under normal light/dark conditions (nLD) and reverse light/dark conditions (rLD). Rats were deprived of water but not food either for 0, 5, 16, 24 or 48 h. As expected, hematocrit ratio (HCT), osmolality (OSM), plasma renin activity (PRA) and weight loss increased as a function of duration of water deprivation. In non-deprived rats (0 h), very little FLI was observed in most brain regions. The number of cells showing FLI increased with duration of water deprivation in the supraoptic nucleus (SON), paraventricular nucleus (PVN), organum vasculosum laminae terminalis (OVLT), median preoptic nucleus (MnPO) and subfornical organ (SFO) in both nLD and rLD conditions. However, the pattern of FLI differed between nLD and rLD conditions. Compared to corresponding nLD groups after 5 or 24-h water deprivation, rLD groups had significantly more FLI in SON and PVN, and higher PRA and HCT. Also, weight loss and FLI in the MnPO were greater after 5 h, and FLI in the SFO was greater after 24 h under rLD compared to nLD conditions. Our findings indicate that the magnitude of c-fos expression, and change in weight and plasma parameters were a function of both the duration of water deprivation and the time of day at which the deprivation started. This may result from ingestion of food early in the deprivation periods during the rLD tests, thus producing greater change in osmolality and blood volume.

Dissociation of Fos-like immunoreactivity in lamina terminalis and magnocellular hypothalamic nuclei induced by hypernatremia.

Rats were given either slow (1 h) or rapid (10 min) intravenous infusions of either 6 or 12 mmol NaCl/kg body weight. Fos-like immunoreactivity (FLI) induced by the infusions was measured in several brain regions. The higher dose of NaCl induced FLI in structures of the lamina terminalis, including organum vasculosum (OVLT) and subfornical organ (SFO), as well as in the magnocellular supraoptic (SON) and paraventricular hypothalamic (PVN) nuclei. The lower dose of hypertonic NaCl induced FLI in only the SON and PVN. Faster delivery of the solute load tended to amplify the FLI in SFO and OVLT. These data confirm and extend previous reports of osmotically-induced FLI in rat brain and demonstrate that the discrepancies between these studies result from different dosage regimens of NaCl. The data are discussed as they relate to the lamina terminalis as a primary osmosensitive region in brain.

Sodium depletion and Fos-immunoreactivity in lamina terminalis.

Rats were either depleted of sodium by treatment with a diuretic or were made hypovolemic by injection of polyethylene glycol (PEG) and their brains were subsequently examined for induction of Fos-like immunoreactivity (FLI). FLI was observed in the lamina terminalis (LT) after both treatments; FLI was observed in the magnocellular neurosecretory hypothalamic areas only after PEG. Implications of these data for brain substrates of sodium appetite are discussed.

Fos induced in brain of spontaneously hypertensive rats by angiotensin II and co-localization with AT-1 receptors.

The induction of Fos-like immunoreactivity (FLI) by peripheral administration of angiotensin II (Ang II) was used to determine whether central activation was greater in spontaneously hypertensive rats (SHR) than in normotensive WKY and outbred Wistar controls. FLI was induced in the same brain regions (circumventricular organs and neurosecretory hypothalamic cell groups) in all three groups of rats, but the FLI in several of these regions was markedly less in WKY than in either SHR or Wistar. This reduced responsiveness in supraoptic and paraventricular nuclei was selective to Ang II, because the FLI induced in these nuclei by hypertonic NaCl did not differ between groups. We also report that a considerable number of cells in the SON and PVH expressing FLI to these stimuli show immunostaining with an antibody to the AT-1 Ang II receptor. These data indicate that central angiotensinergic pathways may be more sensitive in SHR than WKY, and that WKY are less sensitive than outbred Wistars.

Tolerance to the anorectic effect of dexfenfluramine in rats: role of serotonin, cholecystokinin, and neuropeptide Y.

Tolerance to the anorectic effect of dexfenfluramine (DFEN) is shown in rats treated either chronically with low doses or once with a high dose of the agent. Rats given DFEN regimens that result in complete loss of DFEN anorexia showed no change in the anorexia caused by peripheral injection of cholecystokinin (CCK). The orexigenic effects of metergoline and neuropeptide Y were also unaltered as a function of DFEN pretreatment. Both the magnitude and duration of tolerance to a test dose of DFEN seemed to depend, in part, upon contingent (situational) factors, and were independent of whether brain serotonin (5-HT) measures were either unaffected or decreased by the DFEN pretreatment (low and high doses, respectively).

Localization of changes in immediate early genes in brain in relation to hydromineral balance: intravenous angiotensin II.

Immediate early genes, detected by Fos- and Jun-like immunoreactivity (FLI, JLI), were induced in discrete regions of the rat brain by intravenous infusion of angiotensin II (Ang II) at dipsogenic doses. The regions included subfornical organ (SFO), organum vasculosum laminae terminalis (OVLT), median preoptic nucleus (MnPO), supraoptic nucleus (SON), and the magnocellular part of the paraventricular hypothalamus (PVH). These responses were sustained for up to 6 h of infusion. In SFO, FLI was induced preferentially in the posterior part, while JLI occurred in more central regions. Cerebroventricular (ICV) injection of the Ang II type 1 receptor (AT-1) antagonist, losartan potassium, completely prevented the FLI induced by Ang II in these brain regions. ICV injection of the Ang II type 2 receptor (AT-2) antagonist, PD 123319, did not reduce Ang II-induced FLI in SFO, OVLT and MnPO, but markedly attenuated the activation in SON and PVH. To determine whether SFO is the primary site for transduction of the circulating Ang II signal, electrolytic lesions were made in or rostral to the SFO. Rats with complete lesions showed a complete absence of Ang-induced FLI in SON and PVH. The data are discussed in terms of functional mapping of the brain regions activated by circulating Ang II and neural circuitry for water intake, including the possible role of AT-2 receptors in PVH and SON.

Adrenalectomy enhances the anorectic effect of dexfenfluramine: relation to mineralocorticoids and salt appetite.

A previous observation that adrenalectomized (ADX) rats were more anorectic and lost more weight than intact controls when treated with the serotonergic agent, d,l-fenfluramine, was confirmed using a high dose of the more selective d-enantiomer, dexfenfluramine (DFEN). However, ADX rats were not more anorectic than intact controls to a lower dose of DFEN. The extreme weight loss at high doses was accompanied by a loss of appetite for NaCl solution. The rapid deterioration of these animals may have been due to excessive loss of sodium ions and extracellular fluid volume, although this was not assessed directly. This interpretation was supported in a second study in which treatment of ADX rats with a selective mineralocorticoid, which eliminates both the need and appetite for NaCl, normalized weight loss during DFEN treatment. These data do not support the idea of an interactive role of glucocorticoids and brain serotonin in anorexia. The density of serotonin uptake sites in frontal cortex was reduced by DFEN in a dose-dependent manner, but did not differ between ADX and intact control groups.

Effects of glucose and fat antimetabolites on norepinephrine turnover in rat hypothalamus and brainstem.

Rats were injected acutely with antimetabolites of either glucose (2 deoxy-D-glucose, 2DG), fat (methylpalmoxirate, MP or mercaptoacetate, MAC), or the combination of these agents, in dosages known to stimulate food intake. Norepinephrine (NE) turnover in hypothalamus and brainstem was determined after these treatments by the method of synthesis inhibition. Glucoprivation (2DG) increased NE turnover in hypothalamus, confirming previous studies. Fat antimetabolites alone had no effect on NE turnover, nor did a peripherally-acting fructose antimetabolite. Combination of MP and 2DG, but not MAC and 2DG, produced a greater NE turnover than 2DG alone. These data suggest that peripheral signals of metabolic emergency do not activate brain NE systems, except when these systems are already activated by an ongoing cerebral metabolic emergency. The role of hypothalamic NE in metabolic integration of feeding is discussed, and possible hemispheric differences.

Role of renin-angiotensin-aldosterone system in NaCl appetite of rats.

A variety of experimental paradigms is now known to induce an appetite for NaCl solutions in rats. These include 1) bilateral adrenalectomy; 2) hypothyroidism; 3) salivariectomy; and 4) administration of hydrochlorothiazide, metyrapone, estrogen, methylxanthines, captopril, propranolol, large doses of deoxycorticosterone acetate, and intraperitoneal isotonic glucose or subcutaneous polyethylene glycol. A point of commonality among these is that a reduction in preference threshold accompanies the appetite for NaCl in all cases thus far tested. An additional point is the fact that each paradigm inducing a salt appetite, except salivariectomy, can be linked to an effect on the renin-angiotensin-aldosterone system. The level of angiotensin II in the brain may play a role in the induction of a salt appetite in the rat. It is clear, however, that modest doses of mineralocorticoid hormones, given in conjunction with the stimulus producing the salt appetite (e.g., adrenalectomy, thyroidectomy, or treatment with captopril), reduces NaCl intake to control level. Although this effect can be partially explained in most cases by the consequent reduction in angiotensin II production, the salt appetite that occurs when mineralocorticoid concentration in blood is high and angiotensin II concentration is low, or when both are low, requires another explanation. This may be related to the effect of mineralocorticoid hormones on salivary sodium concentration. The role of the concentration of sodium in saliva on intake of NaCl solution provides an alternative explanation for the induction of a salt appetite in rats and merits additional study.