Prostaglandins and hypothalamic neurotransmitter receptors involved in hyperthermia: a critical evaluation.

The role of a prostaglandin of the E series (PGE) in the hypothalamic mechanisms underlying a fever continues to be controversial. This paper reviews the historical literature and current findings on the central action of the PGEs on body temperature (Tb). New experiments were undertaken to examine the local effect of muscarinic, nicotinic, serotonergic, alpha-adrenergic, or beta-adrenergic receptor antagonists at hypothalamic sites where PGE1 caused a rise in Tb of the primate. Guide tubes for microinjection were implanted stereotaxically above sites in and around the anterior hypothalamic, preoptic area (AH/POA) of male Macaque monkeys. Following postoperative recovery, 30-100 ng of PGE1 was micro-injected unilaterally in a volume of 1.0-1.5 microliter at sites in the AH/POA to evoke a rise in Tb, and once identified, pretreated with a receptor antagonist. PGE1 hyperthermia was significantly reduced by microinjections of the muscarinic and nicotinic antagonists, atropine, or mecamylamine, at PGE1 reactive sites in the AH/POA. The serotonergic antagonist, methysergide, injected at PGE1 sensitive sites in the ventromedial hypothalamus also attenuated the rise in Tb. However, the 5-HT reuptake blocker, fluoxetine, and the beta-adrenergic receptor antagonist, propranolol, injected in the AH/POA failed to alter the PGE1 hyperthermia. In contrast, the alpha-adrenergic antagonist, phentolamine, potentiated the increase in Tb at all PGE1 reactive sites in the hypothalamus. An updated model is presented to explain how the concurrent actions of aminergic neurotransmitters acting on their respective receptors in the hypothalamus can interact with a PGE to elicit hyperthermia. Finally, an evaluation of the current literature including recent findings on macrophage inflammatory protein (MIP-1) supports the conclusion that a PGE in the brain is neither an obligatory nor essential factor for the expression of a pyrogen fever.

Alcohol dependence and withdrawal in the rat. An effective means of induction and assessment.

Numerous problems have been associated with previous attempts to develop a suitable method for the induction and assessment of alcohol dependence and withdrawal syndrome in the rat. Using our modification of a common inhalation method for the long-term administration of ethanol, these problems can be eliminated. Adult male rats (Long Evans and Brattleboro) were exposed to ethanol vapor concentrations of 7 to 35 mg/liter of air, which cause rapid development of tolerance and physical dependence. With this inhalation method, it is possible to obtain and easily maintain high levels of ethanol in the blood (150 to 400 mg/dl). When exposure to ethanol is terminated, ethanol is eliminated from the system within 1 to 6 hr. This rapid elimination of ethanol is accompanied by a high susceptibility to withdrawal reactions. The severity of the withdrawal syndrome was assessed within 6 to 24 hr after cessation of the ethanol administration by exposing each rat individually to a 60 to 120-sec period of bell ringing. Convulsive seizures were observed in nearly 90% of the animals tested, with a mortality rate of less than 20%.

Determination of the endogenous and evoked release of catecholamines from the hypothalamus and caudate nucleus of the conscious and unrestrained rat.

The action of catecholamines within the CNS is important for the expression of numerous vegetative and behavioral functions. To understand the role these amines play, it is necessary to measure changes in the levels of these transmitter substances by utilizing new developments and methodology in the behaving animal. Utilizing new developments in methodology, it is possible to measure the release of amines into perfusates obtained from specific sites in the brain of the rat under basal and evoked conditions without prior purification or concentration. Using the push-pull perfusion technique, perfusates were obtained from the hypothalamus and caudate nucleus and analyzed by liquid chromatography with electrochemical detection. It is possible to readily determine basal release of dopamine from the caudate nucleus. Detection of both dopamine and noradrenaline is possible under ephedrine stimulated conditions from both the caudate nucleus and the hypothalamus. Although levels of serotonin (5-HT) were detected in brain perfusates, it may not be of neuronal origin. It may be possible to use these techniques to delineate the roles these amines play in various physiological functions.

Perfusion of vasopressin within the rat brain suppresses prostaglandin E-hyperthermia.

These experiments were undertaken to determine whether arginine vasopressin (AVP) could suppress a prostaglandin hyperthermia and to localize sites of these actions in the rat. Prostaglandin E2 (PGE2) sensitive sites were localized in the ventral-septal area by microinjecting 200 ng/0.5 microliter of prostaglandin E2. During perfusion with an artificial CSF, PGE2 injected into the lateral cerebral ventricle evoked a hyperthermia of more than 1 degree C. Perfusion of 6.5 micrograms/ml of AVP markedly attenuated the PGE2-induced hyperthermia. These results suggest that AVP suppresses PGE2-induced hyperthermia in sites in which PGE2 evokes an increase in core temperature.

Vasopressin: its role in antipyresis and febrile convulsion.

When pyrogenic substances are injected intravenously into experimental animals, a sequence of events is set in motion which involves the hypothalamus and perhaps other portions of the diencephalon to produce a febrile response. We now present evidence that the brain produces its own endogenous antipyretic which may serve as a means of controlling the extent of the fever. When arginine vasopressin is perfused through the lateral septal area of the hypothalamus of the sheep, fever is suppressed. Vasopressin alone does not lower normal body temperature when perfused through this region of the brain. In addition, evidence is provided to indicate that vasopressin is released within the lateral septal area during the febrile response. It is concluded that, in fever, arginine vasopressin may be released in the lateral septal area of the brain and serve as an endogenous antipyretic. Results indicate that, following an initial application of vasopressin into the brain itself, a subsequent similar administration of vasopressin produces seizure-like activity. Therefore, it is suggested that this release of arginine vasopressin may contribute to the production of febrile convulsion.

The role of protein synthesis in the hypothalamic mechanism mediating pyrogen fever.

The effects of inhibition of protein synthesis by anisomycin on the pathogenesis of fever and normal thermoregulatory processes were investigated in the conscious and unrestrained cat. Subcutaneous administration of 5.0-25.0 mg/kg of anisomycin prevented the fever normally evoked by an intravenous infusion of either 1.0 ml (10(8) organisms) of a 1:10 dilution of S. typhosa or 1.0-5.0 ml (3.5 x 10(5)-2.1 X 10(7) cells/ml) of endogenous pyrogen. In addition, systemic pre-treatment with anisomycin delayed and/or blocked the fever typically elicited by a direct micro-injection into the anterior hypothalamic, preoptic area (AH/POA) at AP 12.5-16.0 of 1.0 microliters of the endotoxin. Anisomycin did not alter the hyperthermic response to an anterior hypothalamic injection of either 1.0-7.0 micrograms/1.0 microliters of serotonin (5-HT) or 100.0 ng/1.0 microliters of prostaglandin (PGE). Inhibition of protein synthesis, furthermore, did not prevent the fall in body temperature usually produced by an intrahypothalamic micro-injection of 2.33-14.0 micrograms/1.0 microliters of either norepinephrine (NE) or dopamine (DA). The thermoregulatory capacity of the cat was unaffected by the administration of comparable doses of anisomycin, i.e., the animal was able to maintain normal body temperature (+/- 0.5 degrees C) when exposed to an ambient temperature of either 10 degrees C or 34 degrees C. These results strongly suggest that the synthesis of new protein within the region of the AH/POA is a functional requisite for the development of a pyrogen-induced fever.

Fever produced by intrahypothalamic pyrogen: effect of protein synthesis inhibition by anisomycin.

In the unrestrained cat, the inhibition of protein synthesis by anisomycin, given either subcutaneously (5.0--25.0 mg/kg) or directly into the anterior hypothalamic, preoptic area (1.0--25.0 micrograms) impaired the development of a bacterial fever. S. typhosa infused intravenously (1:10 dilution in 1 ml) or into AH/POA (1.0 microliter) evoked an intense fever which was either significantly delayed or prevented by anisomycin. Conversely, anisomycin failed to affect the typical hyperthermia evoked by 100 ng PGE2 or 1.0--7.0 micrograms 5-HT similarly infused into AH/POA. These data demonstrate that an intermediary humoral factor of unknown nature is required in the hyperthermic effector pathway underlying the febrile response.

Dopamine in the hypothalamus of the cat: pharmacological characterization and push-pull perfusion analysis of sites mediating hypothermia.

Within the rostral diencephalon of the cat, 113 sites were examined for their reactivity to 2.33--14.0 microgram dopamine (DA) or 2.33--14.0 microgram norepinephrine (NE) microinjected in a volume of 0.75 microliter. During each experiment, colonic temperature was monitored and additional physiological measures were recorded continuously. In contrast to CSF controls, an intrahypothalamic injection of either catecholamine at circumscribed sites evoked a dose-dependent fall in the cat's body temperature, with NE ordinarily evoking a more profound hypothermic response. The morphological sites of maximum sensitivity were localized in the anterior hypothalamic, preoptic region. At some but not all sites, a prior microinjection of 3.5--7.0 microgram phentolamine attenuated the magnitude of the DA-induced hypothermia and delayed its onset. Conversely, at all loci, the pretreatment by the injection of this alpha-adrenergic antagonist markedly reduced the absolute magnitude of the NE-induced fall in the cat's temperature. Similar pretreatment of a reactive hypothalamic locus with a beta-adrenergic receptor blocking agent, practolol (3.5 microgram), failed to alter the hypothermia following a microinjection of DA. Either of two DA receptor antagonists, haloperidol (0.04--7.0 microgram) or d-butaclamol (0.48--1.47 microgram), when given in a sufficient dose, effectively delayed the onset of the DA-hypothermia and reduced its absolute magnitude; however, the NE-induced decline in the cat's temperature was unaffected by DA receptor blockade. Endogenous stores of DA and/or NE in the cat's hypothalamus were radio-labeled with either 3H- or 14C-catecholamines or both, microinjected through the implanted guide tube into an identified amine-sensitive site. By using push-pull cannulae, the site was subsequently perfused for 5 min with artificial CSF at a rate of 25 microliter/min with samples collected at 15 min intervals. During either the third or fourth perfusion, the ambient temperature of the cat's chamber of 22--24 degrees C was elevated to 35--45 degrees C and maintained at this level for 15 or 30 min. This environmental warming evoked a release of either DA o; NE or both amines from certain circumscribed sites within the cat's rostral hypothalamus. Overall, these results provide pharmacological, physiological and anatomical evidence for a differential role of DA in the hypothalamic mechanism which mediates the heat loss processes.

Serum thyroxin levels during hyperthermia evoked in the monkey by intrahypothalamic injection of PGE1, 5-HT or pyrogen.

Serotonin (5-HT), prostaglandin E1 (PGE1) or a bacterial pyrogen (E. coli or S. typhosa) was microinjected in a volume of 1.0--1.5 microliter into the hypothalamus of the unanesthetized monkey to evoke a long-term hyperthermia. Samples of venous blood collected every 15 min, before, during and after each fever were analyzed by radioimmunoassay for plasma thyroxin levels. There was no statistically significant correlation between plasma thyroxin values and a given phase of the hyperthermic episode induced by the microinjections of 5--HT, PGE1 or bacteria. The possibility that an enhanced release of the thyroid hormone serves to sustain a long-term elevation in temperature evoked by a centrally acting pyrogenic substance is not supported.

Thermoregulation in the rat: deficits following 6-OHDA injections in the hypothalamus.

Bilateral microinjections of 6-hydroxydopamine (6-OHDA) were made in a volume of 0.5--0.75 microliter through chronically implanted cannulae into anterior hypothalamic, preoptic loci. Sites were selected at which 1.0 to 12.5 microgram of norepinephrine (NE) had previously elicited a fall in the rat's body temperature. After 2.0 to 6.0 microgram of 6-OHDA were injected in the same volume at the same loci, a comparable hypothermia ensued. When the rats were exposed repeatedly for one-hour intervals to an environmental temperature of either 35.0 degrees C or 8.0 degrees C, they were unable to thermoregulate against the heat and their colonic temperature rose. In some experiments, the rats also failed to defend adequately against the cold ambient temperature, but mainly following the microinjection of the higher doses of 6-OHDA . The intakes of food and water were generally suppressed; this was accompanied by a transient decline in body weight. Overall, the severity, duration and direction of the thermoregulatory impairment depended upon the anatomical site of injection and the dose regimen of the neurotoxin employed. These results offer further evidence that an intact catecholaminergic pathway within the anterior hypothalamus is required for the rat's physiological control of heat loss in a warm environmental temperature.