. Activation of spinally projecting and nitrergic neurons in the PVN following heat exposure. Am J Physiol Regul Integr Comp Physiol 291: R91-R101, 2006; doi:10.1152/ajpregu.00675.2005.-The present study investigated the effect of acute thermal stimulation in conscious rats on the production of Fos, a marker of increased neuronal activity, in spinally projecting and nitrergic neurons in the hypothalamic paraventricular nucleus (PVN). The PVN contains a high concentration of nitrergic neurons, as well as neurons that project to the intermediolateral cell column (IML) of the spinal cord that can directly influence sympathetic nerve activity (SNA). During thermal stimulation, the PVN is activated, but it is unknown whether spinally projecting PVN neurons and the nitrergic neurons are involved. Compared with controls, rats exposed to an environmental temperature of 39°C for 1 h had a 10-fold increase in the number of cells producing Fos in the PVN (133 Ϯ 23 vs. 1,336 Ϯ 43, respectively, P Ͻ 0.0001). Of the spinally projecting neurons in the PVN of heated rats (98 Ϯ 10), over 20% expressed Fos. Additionally, of the nitrergic neurons (NADPH-diaphorase positive) located in the parvocellular PVN (723 Ϯ 17), ϳ40% also expressed Fos (P Ͻ 0.0001 compared with controls). Finally, there was a significant increase in the number of spinally projecting neurons in the PVN that were nitrergic and expressed Fos after heat exposure (12%) compared with controls (0.1%) (P Ͻ 0.0001). These results suggest that spinally projecting and nitrergic neurons in the PVN may contribute to the central pathways activated by thermal stimulation. Fos immunohistochemistry; spinally projecting EXPOSURE TO A HOT TEMPERATURE challenge elicits responses mediated in part by the autonomic nervous system to promote heat loss and maintain body fluid homeostasis. Such responses include sweating, an increase in heart rate, increased respiration rate, skin vasodilation, and visceral vasoconstriction in humans; increased salivary secretion in rodents; and tail vasodilation in rats (25,27). The latter autonomic cardiovascular responses often involve changes in sympathetic nerve activity (SNA) and result in the redistribution of blood flow from the viscera to the skin. These changes are mediated by the central nervous system (CNS) (24,25,40,43,45,52,65).It has been well established that the CNS is essential in the regulation of body temperature. There are several brain regions that are likely to contribute to the CNS pathways that mediate the thermoregulatory responses. Studies using the marker of neuronal activation, Fos, or electrophysiological recordings have shown that several forebrain areas are activated after the elevation in body temperature (2, 5-7, 21, 28, 34, 38, 39, 42, 52, 53). These forebrain areas include the preoptic area, anterior hypothalamus, and the paraventricular nucleus (PVN) of the hypothalamus. The preoptic area and anterior hypothalamus are well known key thermoregulatory sites within the brain; however, a role of the PVN in thermoreg...

The rostral ventrolateral medulla (RVLM) is essential for the generation of sympathetic nerve activity. The RVLM receives a substantial innervation from the hypothalamic paraventricular nucleus (PVN). Activation of P2X purinoceptors via ATP has been shown to mediate fast excitatory synaptic neurotransmission. There is mounting evidence to suggest the presence of P2X purinoceptors in hypothalamic nuclei, including the PVN. In this study, we determined whether P2X 1 -P2X 6 purinoceptor subtypes were present on PVN neurones that projected to the RVLM. Injection of the retrogradely transported tracer, rhodamine-tagged microspheres, into the pressor region of the RVLM was used to identify the neurones in the PVN that innervated the RVLM. P2X 1 -P2X 6 purinoceptors were detected by immunohistochemistry. Double-labelled neurones were quantified and expressed as a proportion of the retrogradely labelled neurones. The proportions of double-labelled neurones for each of the P2X purinoceptor subtypes varied, on average, from 14 to 29%. The P2X 3 purinoceptor subtype was found to be the dominant purinoceptor subtype present on PVN neurones projecting to the RVLM. Additionally it was apparent that more than one P2X purinoceptor subtype was present on the PVN neurones projecting to the RVLM, since the sum of the average percentages of double-labelled neurones for each P2X purinoceptor subtype exceeded 100%. These findings highlight the presence of the P2X 1 -P2X 6 purinoceptors on PVN neurones projecting to the RVLM. The results suggest a potential role for ATP in the PVN in the regulation of sympathetic nerve activity.

Redistribution of blood from the viscera to the peripheral vasculature is the major cardiovascular response designed to restore thermoregulatory homeostasis after an elevation in body core temperature. In this study, we investigated the role of the hypothalamic paraventricular nucleus (PVN) in the reflex decrease in renal blood flow that is induced by hyperthermia, as this brain region is known to play a key role in renal function and may contribute to the central pathways underlying thermoregulatory responses. In anesthetized rats, blood pressure, heart rate, renal blood flow, and tail skin temperature were recorded in response to elevating body core temperature. In the control group, saline was microinjected bilaterally into the PVN; in the second group, muscimol (1 nmol in 100 nl per side) was microinjected to inhibit neuronal activity in the PVN; and in a third group, muscimol was microinjected outside the PVN. Compared with control, microinjection of muscimol into the PVN did not significantly affect the blood pressure or heart rate responses. However, the normal reflex reduction in renal blood flow observed in response to hyperthermia in the control group ( approximately 70% from a resting level of 11.5 ml/min) was abolished by the microinjection of muscimol into the PVN (maximum reduction of 8% from a resting of 9.1 ml/min). This effect was specific to the PVN since microinjection of muscimol outside the PVN did not prevent the normal renal blood flow response. The data suggest that the PVN plays an essential role in the reflex decrease in renal blood flow elicited by hyperthermia.

A major integrative site within the brain for autonomic function is the hypothalamic paraventricular nucleus (PVN). Several studies have suggested that the PVN may be involved in the responses regulating body temperature. Hyperthermia elicits redirection of blood flow from the viscera to the periphery and involves changes in sympathetic nerve activity mediated by the central nervous system. The hypothalamic PVN includes neurones that project to the rostral ventrolateral medulla (RVLM), an important autonomic region involved in the tonic regulation of sympathetic nerve activity. This pathway could contribute to the cardiovascular changes induced by hyperthermia. The PVN has a high concentration of nitrergic neurones and it is known that nitric oxide within the brain mediates heat dissipation. Thus the aims of this study were to determine whether RVLM-projecting neurones in the PVN are activated by heat and whether those neurones are also nitrergic. The results show that, compared with control conditions, exposure of conscious rats to a hot environment of 39• C significantly increased the number of neurones containing a Fos-positive nucleus (a marker of activation) and significantly increased the number of activated RVLM-projecting neurones in the PVN. Also, although heating significantly increased the number of activated nitrergic PVN neurones, triple-labelled neurones (i.e. activated, nitrergic and RVLM projecting) in the PVN were rarely observed. The results suggest that RVLM-projecting neurones in the PVN may play a role in responses to heat exposure but these are not nitrergic.

In the present study, we investigated whether the PVN is a key region in the mesenteric vasoconstriction that normally accompanies an increase in core body temperature. Anesthetized rats were monitored for blood pressure, heart rate, mesenteric blood flow, and vascular conductance. In control rats, elevation of core body temperature to 41°C had no significant effect on blood pressure, increased heart rate, and reduced mesenteric blood flow by 21%. In a separate group of rats, muscimol was microinjected bilaterally (1 nmol/side) into the PVN. Compared with the control group, there was no significant difference in the blood pressure and heart rate responses elicited by the increase in core body temperature. In contrast to control animals, however, mesenteric blood flow did not fall in the muscimol-treated rats in response to the elevation in core body temperature. In a separate group, in which muscimol was microinjected into regions outside the PVN, elevating core body temperature elicited the normal reduction in mesenteric blood flow. The results suggest that the PVN may play a key role in the reflex decrease in mesenteric blood flow elicited by hyperthermia.reflex mesenteric blood flow; increased core body temperature CARDIOVASCULAR RESPONSES TO changes in body temperature include marked changes in sympathetic nerve activity and blood flow. In response to hyperthermia, for example, there are increases in splanchnic, renal, splenic, and lumbar sympathetic nerve activity (25,28), resulting in vasoconstriction of the respective vascular beds. In contrast, hyperthermia elicits vasodilation in the skin vasculature, and in the rat tail vasodilation also occurs (21, 24); the tail is a major thermoregulatory organ in the rat. These changes contribute to the redistribution of the blood flow during hyperthermia such that blood is redirected from the warm internal environment in visceral organs to the peripheral vasculature where heat can be dissipated.The alterations in blood flow, mediated by changes in sympathetic nerve activity, involve the central nervous system. The sites in the brain that are likely to be involved in mediating the responses include the sympathetic premotor nuclei located in the hypothalamus and brain stem. These regions contain neurons that directly project to the sympathetic preganglionic motor neurons located in the intermediolateral cell column of the thoracolumbar spinal cord.Within the forebrain, the hypothalamic paraventricular nucleus (PVN) is a key integrative site involved in hormonal, endocrine, and neural control. The PVN is composed of different neuronal subgroups subserving different functions, including cardiovascular regulation. The PVN is one of the small number of premotor nuclei present in the brain (4, 47). There are neurons in the PVN that project to regions of the spinal cord where sympathetic preganglionic neurons are located and thereby can directly influence sympathetic activity (8,40,48). In addition to the spinal projecting neurons present in the PVN, there are neuronal sub...