Effects of adrenalectomy, propranolol and methylatropine on the increase in heart rate induced by injection of dermorphin in the rat anterior hypothalamic nucleus

We have examined, at the light microscope level, putative direct synaptic interconnections, via motor axon collaterals, between type-identified triceps surae alpha-motoneurons labeled by intracellular injection of HRP. The results indicate that monosynaptic recurrent contacts can occur between synergist motoneurons (in this case, medial gastrocnemius to soleus, and vice versa), irrespective of motor unit type (type FF to type S, and vice versa).     

Increases in heart rate and blood pressure produced by injections of dermorphin into discrete hypothalamic sites

Single unit electrical activity has been recorded from dorsal horn neurones in the lumbar spinal cord of adult rats which had been treated at birth with either capsaicin (50 mg kg-1) or with the solvent-vehicle only. The responses of these neurones to electrical stimulation of A- and C-fibres in the sural nerve and to natural stimulation of their cutaneous receptive fields have been studied. In vehicle-injected rats, 54% of the units driven by electrical stimulation of the A-fibres in the sural nerve could also be driven by stimulation of the C-fibres in this nerve. In capsaicin-treated animals, only 30% of such units had a C-fibre input from the sural nerve. In vehicle-injected rats, 51.5% of the neurones with a C-fibre input showed a 'wind-up' effect on repetitive C-fibre stimulation of the sural nerve at 1 Hz. A similar proportion of neurones (55%) displayed this effect in capsaicin-treated rats. There were fewer neurones with very intense 'wind-up' in capsaicin-treated compared to vehicle-treated rats. In capsaicin-treated animals, greater proportions of neurones with 'wind-up' were superficially located in the dorsal horn, had small receptive fields and were driven only by cutaneous nociceptors. The proportions of neurones driven by innocuous mechanical stimulation of the skin, by noxious mechanical stimulation or by both forms of stimulation were similar in vehicle-injected and capsaicin-treated animals. In capsaicin-treated rats, more neurones had 'medium-sized' receptive fields than in vehicle-injected rats. In capsaicin-treated rats, more neurones had receptive fields in the foot and ankle than in vehicle-injected animals, where receptive fields in the toes were predominant. Some neurones showed expanded receptive fields after repetitive electrical stimulation of C-fibres at 1 Hz. This expansion occurred more often in neurones recorded from capsaicin-treated animals than in those of vehicle-injected rats. These results are discussed in relation to the role of afferent C-fibres in sensory mechanisms.     

Effects of adrenalectomy on CART expression in the rat arcuate nucleus

In order to test for glucocorticoid regulation of CART in the arcuate nucleus, adrenalectomies (ADX) and hormone replacements (HRs) were carried out in groups of rats. CART mRNA levels were determined by in situ hybridization and CART peptide levels by immunocytochemistry. ADX caused a lowering of CART mRNA and peptides levels in the arcuate and this was reversed by HR. These results indicate a glucocorticoid regulation of CART in the arcuate. The regulation could be direct through an action of glucocorticoid receptors or indirectly through ADX-induced changes in leptin levels. These findings suggest a role for CART in the stress response.     

Convergence of thermal, osmotic and cardiovascular signals on preoptic and anterior hypothalamic neurons in the rat

Responsiveness of thermosensitive neurons in the preoptic and anterior hypothalamus (PO/AH) to osmotic and cardiovascular signals have been shown to be responsible, at least partly, for the reduced thermoregulation during dehydration and the hypothermia after acute blood loss. The responsiveness to local and peripheral (hepatoportal) osmotic stimuli were found in about 60% of PO/AH thermosensitive neurons and 12% of thermally insensitive neurons in tissue slices in vitro and in urethane-anesthetized rats. Since hyperosmotic stimuli predominantly decreased the activity of both warm-sensitive and cold-sensitive neurons, the reduced heat loss and heat production during dehydration may be explained by altered activity of PO/AH thermosensitive neurons induced by hyperosmolality. About 42% of 250 PO/AH neurons (66.3% of thermosensitive neurons and 30% of thermally insensitive neurons) exhibited the responsiveness to changes in blood pressure by less than 15 mmHg, which was found to be mediated by baro/volume receptors. Hypotensive stimuli predominantly increased the activity of warm-sensitive neurons and decreased the activity of cold-sensitive neurons. The neuronal responses may explain, at least in part, the hypothermia after acute bleeding.     

Organization of cortical, basal forebrain, and hypothalamic afferents to the parabrachial nucleus in the rat.

In a previous study (Herbert et al., J. Comp. Neurol. [1990];293:540-580), we demonstrated that the ascending afferent projections from the medulla to the parabrachial nucleus (PB) mark out functionally specific terminal domains within the PB. In this study, we examine the organization of the forebrain afferents to the PB. The PB was found to receive afferents from the infralimbic, the lateral prefrontal, and the insular cortical areas; the dorsomedial, the ventromedial, the median preoptic, and the paraventricular hypothalamic nuclei; the dorsal, the retrochiasmatic, and the lateral hypothalamic areas; the central nucleus of the amygdala; the substantia innominata; and the bed nucleus of the stria terminalis. In general, forebrain areas tend to innervate the same PB subnuclei from which they receive their input. Three major patterns of afferent termination were noted in the PB; these corresponded to the three primary sources of forebrain input to the PB: the cerebral cortex, the hypothalamus, and the basal forebrain. Hypothalamic afferents innervate predominantly rostral portions of the PB, particularly the central lateral and dorsal lateral subnuclei. The basal forebrain projection to the PB ends densely in the external lateral and waist subnuclei. Cortical afferents terminate most heavily in the caudal half of the PB, particularly in the ventral lateral and medial subnuclei. In addition, considerable topography organization was found within the individual projections. For example, tuberal lateral hypothalamic neurons project heavily to the central lateral subnucleus and lightly to the waist area; in contrast, caudal lateral hypothalamic neurons send a moderately heavy projection to both the central lateral and waist subnuclei. Our results show that the forebrain afferents of the PB are topographically organized. These topographical differences may provide a substrate for the diversity of visceral functions associated with the PB.     

Effects of endogenous sugar acids on the ventromedial hypothalamic nucleus of the rat

Effects of certain endogenous sugar acids such as 2-deoxytetronic acid (2-DTA) and 3-deoxypentonic acid (3-DPA) were investigated on the neuronal activity of the ventromedial hypothalamic nucleus (VMH) in the rat. Electrophoretically applied 2-DTA significantly and specifically facilitated the activity of glucoreceptor neurons in the VMH, while 3-DPA suppressed their activity. However, non-glucoreceptor neurons were not affected by these sugar acids. These results reinforces the previous suggestion that 2-DTA and 3-DPA also might act as endogenous satiety and hunger substances, respectively.     

APJ receptor mRNA expression in the rat hypothalamic paraventricular nucleus: regulation by stress and glucocorticoids

The apelin receptor (APJ receptor, APJR) has recently come to prominence following the isolation and identification of its endogenous ligand, apelin, from bovine stomach tissue extracts. Investigation of APJR mRNA expression has revealed a hypothalamic distribution similar to that of vasopressin suggesting that the apelin-APJR system may be involved in the regulation of the hypothalamic-adrenal-pituitary (HPA) stress axis. To investigate whether APJR is involved in the regulation of hypothalamic function during stress, APJR mRNA expression levels were measured by in situ hybridization in the hypothalamus of rats subjected to acute and repeated restraint stress. Acute stress caused an increase in APJR mRNA expression in the hypothalamic parvocellular paraventricular nucleus (pPVN) while repeated restraint stress induced a sustained up-regulation of pPVN APJR mRNA expression in intact rats. Removal of endogenous glucocorticoids by adrenalectomy also resulted in an increased expression of APJR mRNA in the PVN, suggesting a negative regulation of APJR mRNA expression by glucocorticoids. The role of glucocorticoids in mediating these stress-induced changes was investigated by analysing the effects of acute and repeated restraint stress on APJR mRNA levels in adrenalectomized rats. In these rats, APJR mRNA expression levels did not change above the already elevated levels of adrenalectomized-control rats. These data suggest that acute and repeated stress exert a stimulatory influence on APJR mRNA expression at the hypothalamic level that may be dependent on basal levels of circulating glucocorticoids, and further suggest a role for APJR in the regulation of hypothalamic function.     

Hypothalamic-immune interactions. I. The acute effect of anterior hypothalamic lesions on the immune response

Rats with electrolytic anterior hypothalamic lesions show changes in lymphoid tissue cellularity and a decrease in the response to concanavalin A (Con A). This effect manifests itself maximally 4 days after lesioning, with a return to normal by day 14. The changes are not mediated through the release of corticosteroids. These data indicate the presence of a neuroendocrine pathway that is capable of modulating immune function.     

Glucosensing in parvocellular neurons of the rat hypothalamic paraventricular nucleus

Specialized hypothalamic neurons responding to rising extracellular glucose via increases or decreases in their electrical activity [glucose-excited (GE) and glucose-inhibited (GI) cells, respectively] have been reported in the hypothalamic arcuate, ventromedial and lateral nuclei. The hypothalamic paraventricular nucleus (PVN) is an important neurosecretory and preautonomic output nucleus. We tested whether parvocellular PVN neurons also possess glucosensing properties, using patch-clamp recording and immunocytochemistry. Putative neurosecretory (p-NS) and preautonomic (p-PA) cells were identified electrophysiologically. Although parvocellular neurons were insensitive to transitions from 10 to 2.5 mm glucose, approximately 68% of p-PA cells responded directly to glucopenia (mimicked by a step to 0.2 mm glucose) with an increased membrane conductance. Of these, approximately 24% hyperpolarized (accompanied by an outward current) and thus were GE, approximately 26% depolarized (with an inward current, thus GI) and approximately 18% did not change membrane potential. The concentration dependence of the glucose response was similar for both GE and GI cells (EC(50) of 0.67-0.7 mm), but was steep, with Hill slopes of 3-4. The K(ATP) channel blockers glibenclamide and tolbutamide did not prevent, while the K(ATP) channel opener diazoxide did not mimic, the effects of low glucose on GE neurons. Moreover, the K(ATP) sulfonylurea receptor SUR1 was not detected in glucosensitive neurons. We conclude that the PVN contains previously unknown GE and GI cells that could participate in regulation of autonomic functions. GE neurons in the PVN sense ambient glucose via a unique mechanism, probably independent of K(ATP) channels, in contrast to neurons in other hypothalamic nuclei.     

Activation of the rat hypothalamic supramammillary nucleus by food anticipation,food restriction or ghrelin administration

The circulating orexigenic hormone ghrelin targets many brain areas involved in feeding control and signals via a dedicated receptor, the growth hormone secretagogue receptor 1A. One unexplored target area for ghrelin is the supramammillary nucleus (SuM), a hypothalamic area involved in motivation and reinforcement and also recently linked to metabolic control. Given that ghrelin binds to the SuM, we explored whether SuM cells respond to ghrelin and/or are activated when endogenous ghrelin levels are elevated. We found that peripheral ghrelin injection activates SuM cells in rats, reflected by an increase in the number of cells expressing c‐Fos protein in this area, as welll as by the predominantly excitatory response of single SuM cells recorded in in vivo electrophysiological studies. Further c‐Fos mapping studies reveal that this area is also activated in rats in situations when circulating ghrelin levels are known to be elevated: in food‐restricted rats anticipating the consumption of food and in fed rats anticipating the consumption of an energy‐dense food. We also show that intra‐SuM injection of ghrelin induces a feeding response in rats suggesting that, if peripheral ghrelin is able to access the SuM, it may have direct effects on this brain region. Collectively, our data demonstrate that the SuM is activated when peripheral ghrelin levels are high, further supporting the emerging role for this brain area in metabolic and feeding control.     

Distribution of neurons in the anterior hypothalamic nucleus activated by blood pressure changes in the rat

Electrophysiological and Fos-like protein immunocytochemical methods were used to identify the number and distribution of anterior hypothalamic neurons that are activated by changes in arterial pressure. First, in anesthetized, male Sprague-Dawley rats, arterial pressure increases and decreases led to differential activation of neurons in the anterior hypothalamic nucleus. Most of the units that responded to a rise in arterial pressure with a decrease in activity (pressor units) were located in the central part of the anterior hypothalamic nucleus, whereas units that increased firing when arterial pressure rose (the depressor units) were found throughout the nucleus. Second, in awake, male Sprague-Dawley rats, Fos-like protein immunoreactivity was mapped following sustained arterial pressure changes. Within the anterior hypothalamus, reduction in arterial pressure increased the number of Fos-labeled neurons primarily in the paraventricular nucleus and to a lesser extent in the anterior half of the anterior hypothalamic nucleus. In contrast, elevation in arterial pressure increased Fos labeling throughout the anterior hypothalamic nucleus and to a lesser extent in the paraventricular nucleus.     

Sexual dimorphism in the preoptic/anterior hypothalamic area of ferrets: Effects of adult exposure to sex steroids

The organization of neuronal cell bodies in the caudal preoptic area (POA) and rostral anterior hypothalamic area (AH) was studied in Nissl-stained brain sections from adult male and female ferrets. Computer-assisted image-analysis procedures were developed to help estimate the areas of cellular density and the sizes of individual perikarya. At the junction of the POA and AH, a bilateral dorsal-medial group of neurons was apparent only in male ferrets (dorsal nucleus). At the same coronal level, a ventral-medial group of neurons was apparent bilaterally in both males and females (ventral nucleus). The mean somal area of cells in the dorsal nucleus of males was significantly greater than the mean somal area of cells in the corresponding dorsal region of females or in the ventral nucleus of both sexes. The dorsal nucleus was clearly discernible in adult males regardless of their hormonal status, although cells in the dorsal nucleus were larger in intact breeding males or gonadectomized males given testosterone, estradiol or dihydrotestosterone than in gonadectomized males given no gonadal hormones or given progesterone. Neither the grouping of large cells nor the steroid-induced increase in cell size, characteristic of the male dorsal nucleus, was seen in the comparable dorsal region of females. The sex difference in cellular organization observed in the ferret at the junction of the POA and AH is the first difference of this type to be seen in the POA/AH of a non-rodent mammalian species. Its identification will, hopefully, aid in the analysis of the neural mechanisms that control various sex-specific behaviors in this species.     

Activity of ventromedial hypothalamic neurons suppressing heart rate is associated with paradoxical sleep in the rat

Cardiovascular change is one of the common features of paradoxical sleep. Our study offers evidence that one of the central areas regulating the circulation during sleep is the ventromedial nucleus of the hypothalamus (VMH). We found a group of neurons in this hypothalamic nucleus of rats whose electrical activity was exclusively increased during paradoxical sleep, and was associated with a reduction in heart rate. The onset of this neural activity usually followed that of paradoxical sleep. The incidence and duration of paradoxical sleep was increased by means of microinjection of carbachol, a cholinergic agonist, into the pontine reticular formation, and the neural activity of the VMH still appeared in synchrony with carbachol-induced paradoxical sleep. These results suggest that the cholinergic paradoxical sleep-inducing mechanism in the pons facilitate the excitability of these neurons. We have previously shown that these VMH neurons suppress blood pressure and heart rate via inhibition of the vasomotor neurons in the medulla oblongata. Taken together, our findings suggest that a group of neurons in the VMH suppresses the circulatory system during paradoxical sleep.     

Inhibitory and facilitatory influences on mating in the female rat effected by lesions of the anterior hypothalamus or the preoptic area

The effects of lesions of the anterior hypothalamus and preoptic area on mating behavior, estrous cyclicity and ovulation in the female rat were evaluated. (1) Significant levels of mating behavior occurred after ovariectomy, during vaginal diestrus, in $\text{4}\text{8}$ animals with lesions of the anterior hypothalamus, and in $\text{2}\text{8}$ animals with lesions of the preoptic area. (2) The anterior hypothalamic and preoptic area lesioned groups showed more mating behavior than the respective sham groups following injection of the lowest dose of estrogen. (3) Lesions of the anterior hypothalamus failed to reduce mating behavior on days of proestrus, and increased mating on days of estrus compared with sham-anterior hypothalamic lesions. (4) Lesions of the preoptic area reduced mating behavior on days of proestrus, but increased mating on days of estrus compared with sham-preoptic area lesions (5) Lesions of the anterior hypothalamus, and shamlesions of the anterior hypothalamus or preoptic area did not disrupt estrous cycles, or reduce the number of tubal ova present on a morning of estrus. (6) Lesions of the preoptic area were more likely to disrupt cyclicity; they prolonged vaginal estrus in $\text{3}\text{8}$ animals, and prevented ovulation and development of pseudopregnancy in $\text{4}\text{8}$ animals. (7) The data suggest that lesions of the anterior hypothalamus interfered with inhibitory influences on mating without disrupting estrous cycles or ovulation. (8) Lesions of the preoptic area interfered with both facilitatory and inhibitory influences on mating behavior, and were more likely to disrupt estrous cycles, ovulation and the development of pseudopregnancy.     

Neurotensin in the rat anterior pituitary gland: effects of endocrinological manipulations

Neurotensin-like immunoreactivity was detected by radioimmunoassay in the anterior lobe of the pituitary gland of several mammalian species, including man. In the rat, pituitary stalk transection did not change the content of neurotensin-like immunoreactivity. Surgical and chemical thyroidectomy produced a drastic reduction of neurotensin-like immunoreactivity in the anterior pituitary gland, whereas adrenalectomy and gonadectomy were without an effect and the decrease consequent to thyroidectomy was restricted to the pituitary gland. These results suggest an interaction between neurotensin and the hypothalamus/pituitary/thyroid axis at the level of the anterior pituitary gland.     

Neuronal control of the ventromedial nucleus from the periventricular nucleus in the hypothalamic slice of the rat

A neuronal pathway between periventricular nucleus and ventromedial hypothalamus (VMH) was investigated by both anatomical and electrophysiological methods. Horseradish peroxidase (HRP)-labelled clusters were observed in the periventricular nucleus after application of HRP into the VMH. Periventricular nucleus stimulation produced mainly excitation of VMH neurones with a latency of 3 msec. These results suggest that periventricular nucleus sends information in the cerebro-spinal fluid to the VMH.     

Paraventricular-coerulear interactions: role in hypertension induced by prenatal undernutrition in the rat

Rats submitted to fetal growth retardation by in utero malnutrition develop hypertension when adult, showing increased hypothalamic mRNA expression for corticotropin-releasing hormone (CRH) and increased central noradrenergic activity. As hypothalamic CRH serves as an excitatory neurotransmitter within the locus coeruleus (LC) and coerulear norepinephrine plays a similar role within the paraventricular nucleus (PVN) of the hypothalamus, we studied, in both normal and prenatally undernourished 40-day-old anesthetized rats, the effects of intra-LC microinjection of CRH and intra-PVN microinjection of the alpha(1)-adrenoceptor antagonist prazosin on multiunit neuronal activity recorded simultaneously from the two nuclei, as well as the effects on systolic pressure. Undernutrition was induced during fetal life by restricting the diet of pregnant mothers to 10 g daily, whereas mothers of control rats received the same diet ad libitum. At day 40 of postnatal life: (i) undernourished rats showed increased neuronal activity in the PVN and LC, as well as increased systolic pressure; (ii) intra-LC CRH stimulated LC and PVN neurons and increased systolic pressure only in normal rats; (iii) intra-PVN prazosin decreased LC and PVN neuronal activity and systolic pressure only in undernourished rats; and (iv) in normal rats, prazosin prevented the stimulatory effect of CRH only in PVN activity; in undernourished rats, prazosin allowed CRH to regain its stimulatory effects. The results point to the existence of an excitatory PVN-LC closed loop, which seems to be hyperactive in prenatally undernourished rats as a consequence of fetal programming; this loop could be responsible, in part, for the hypertension developed by these animals.     

Effects of intraventricular norepinephrine on preoptic-anterior hypothalamic electrical activity in the freely-moving rat: modulation by ovarian steroid hormones

In freely-moving female rats the effects of intraventricular infusion of norepinephrine (NE) on multiunit activity (MUA) were examined in the Diagonal Band of Broca (DBB), medial preoptic area (MPOA) and anterior hypothalamic area (AHA), regions containing neurons producing luteinizing hormone-releasing hormone (LHRH). NE was infused at a dosage known to depress plasma LH levels in ovariectomized (OVX) rats and elevated plasma LH concentrations in OVX-estrogen-progesterone-primed (EBP) animals. It was found that in adult OVX rats MUA in the brain areas listed above was almost invariably inhibited by NE (20 of 23 cases or 87%; the other three cases showed no change in electrical activity). However, after priming with estrogen and progesterone only about one-third of the OVX-EBP rats gave an inhibitory response, with another one-third showing no change in MUA and the final one-third of the cases actually giving an excitatory MUA response to NE — the DBB neurons being the most positive in this regard. Thus it appears that responsiveness of LHRH and/or adjacent neurons to the modulatory action of NE may itself be modulated by the influence of gonadal steroids.     

Effect of anterior hypothalamic and mammillary area lesions on territorial aggressive behaviour in male rats

Adult male Wezob-rats were bilaterally lesioned in either the medial anterior hypothalamic area or the mammillary bodies. The behaviour of these animals when confronted with a male intruder within their own territory, was observed and recorded before and after lesioning and compared with the behaviour of sham-operated animals.Anterior hypothalamic lesions, including large parts of the anterior hypothalamus, the rostral part of the ventromedial hypothalamic nucleus and smaller caudal parts of the preoptic area, led to strong increases in defensive behaviour. This included a decreased tendency to investigate the intruder and an exaggerated defensive reaction when approached by the intruder. Ingestive behaviour and bodyweight were enhanced.Mammillary body lesions, including large parts of the ventral and dorsal premammillary nucleus, the caudal part of the arcuate nucleus, the medial mammillary nucleus, the posterior mammillary nucleus, the supramammillary peduncle and closely surrounding areas, led to a marked increase in offensive behaviour. This was characterized by high levels of initatives and aggression towards an intruder.It is suggested that two distinct neural substrates exist in the medial hypothalamus, which normally modulate defensive (anterior medial hypothalamus) or offensive (posterior medial hypothalamus) aspects of intermale aggression.