Activity of thermosensitive neurons of monkey preoptic hypothalamus during thermoregulatory operant behavior

Unit activities of thermosensitive (TS) and non-TS neurons in the medial preoptic area (MPO) of the monkey were investigated during a high fixed-ratio (FR 12-30) bar press behavior to seek cool air (CT) in a warm environment. The monkey performed the task when ambient temperature (Ta) was raised (35-43 degrees C) and the animal was rewarded with a fall in Ta (23-25 degrees C). The population of neurons which changed the activity at least in one phase of CT was significantly higher among TS neurons (42 of 46) than among non-TS neurons (29 of 64). The most frequently observed responses were the sustained increase or decrease in activity during the bar press period and/or the cooling period. The bar press related activity was not a pure motor coupled response, but its magnitude was greatly modified by the strength of motivation of animals to seek cool air. The cooling phase-related activity was not a thermosensory response which simply reflects the level of skin temperature or its change. It was shown that the response was dependent upon the rewarding value of the cooling air. The results suggest that TS neurons in the MPO may be involved in the control of thermoregulatory cooling behavior.     

Preoptic/anterior hypothalamic neurons: thermosensitivity in wakefulness and non rapid eye movement sleep

Thermosensitive neurons of the preoptic/anterior hypothalamic area (POAH) have been implicated in the regulation of both body temperature and non rapid eye movement (NREM) sleep. During NREM sleep, a majority of POAH warm-sensitive neurons (WSN) exhibit increased discharge compared to wakefulness. Cold-sensitive neurons (CSN) exhibit reduced discharge in NREM sleep compared to wakefulness. To further study the mechanism underlying these processes, the present study compared discharge rate and thermosensitivity (discharge rate change/°C) of WSNs and CSNs in NREM sleep and wakefulness in freely moving adult cats. The thermosensitivity of 24 WSNs and 31 CSNs from the medial POAH was determined from responses to local POAH warming and cooling. WSNs with increased discharge in NREM sleep exhibited increased thermosensitivity during NREM sleep compared to wakefulness. CSNs with decreased discharge during NREM sleep exhibited decreased thermosensitivity in NREM sleep. The change in thermosensitivity from wakefulness to NREM sleep was correlated with the change in discharge rate in WSNs but not in CSNs. In addition, 9 of 47 neurons that were thermo-insensitive during wakefulness became warm-sensitive during NREM sleep. Changes in POAH neuronal thermosensitivity could be a component of the mechanism for stabilization of state after state transition.     

Sleep-active neurons in the preoptic area project to the hypothalamic paraventricular nucleus and perifornical lateral hypothalamus

The lamina terminalis consists of the organum vasculosum of the lamina terminalis (OVLT), median preoptic nucleus (MnPO) and subfornical organ. The MnPO and ventrolateral preoptic area (vlPOA) are known to contain high densities of neurons that are sleep active. The prevalence of sleep-active neurons in the OVLT and subfornical organ is unknown. The vlPOA and subdivisions of the lamina terminalis project to hypothalamic regions involved in the control of behavioral, electrographic or autonomic arousal, including the lateral hypothalamic area (LHA) and paraventricular nucleus (PVN). The extent to which projection neurons are active during sleep is unknown. We quantified c-Fos protein immunoreactivity (IR) in the lamina terminalis and vlPOA in sleeping and awake rats that received injections of retrograde tracer into either the LHA or PVN. Fos IR was also examined in lamina terminalis neurons following tracer injections into the vlPOA. Significantly more projection neurons from the MnPO, OVLT and vlPOA to the LHA were Fos-immunoreactive in sleeping vs. awake animals. Waking Fos IR was more prevalent in lamina terminalis neurons projecting to the PVN although a subset of MnPO projection neurons in sleeping rats was Fos-immunoreactive. Almost 50% of vlPOA-PVN projection neurons expressed Fos IR during sleep, compared with 3% during waking. Significantly more neurons in the OVLT and MnPO projecting to the vlPOA were Fos-immunoreactive in sleeping vs. awake rats. Inhibition of LHA and PVN neurons arising from OVLT, MnPO and vlPOA neurons may contribute to suppression of behavioral, electroencephalographic and sympathetic nervous system activation during sleep.     

Responses of preoptic thermo sensitive neurons to mediobasal hypothalamic stimulation

HORI, T., T. KIYOHARA, T. OSAKA, M. SHIBATA AND T. NAKASHIMA. Responses of preoptic thermosensitiveneurons to mediobasal hypothalamic stimulation. BRAIN RES. BULL. 8(6) 677–683, 1982.—Single-unit responses of preoptic-anterior hypothalamic (PO/AH) thermosensitive neurons to electrical stimulation of the mediobasal hypothalamus (MBH: ventromedial and arcuate nuclei) have been studied in urethane-anesthetized male rats. About half of thermosensitive neurons (46.4% of 69 warm-units and 50% of 28 cold-units) were initially inhibited following MBH stimulation with latencies up to 51.2 msec, while only 29.7% of 91 thermally insensitive units showed the inhibitory response. Orthodromic excitation of spontaneous activity was observed in 17.6% of the units (13.0% of warm-units, 14.3% of cold-units and 21.9% of insensitive units) with the onset latencies between 1.0 and 41.3 msec. A small proportion of neurons (5.3%) were antidromically activated by MBH stimulation, and the estimated conduction velocities of these neurons ranged from 0.11 to 0.67 m/sec. The results suggest reciprocal connections between PO/AH thermosensitive neurons and MBH, which may play some roles in thermoregulation and, also, in functional interactions between thermoregulation and other vital functions controlled in the hypothalamus.     

Changes in dendritic spine density in the preoptic area of the female rat at puberty

Preoptic area (POA) neurons stained by a modification of the Golgi-Kopsch method were studied in developing female rats. Particular attention was focused on dendritic spine densities as they might be altered in relation to the onset of puberty. Neurons in the anterior superior and posterior superior portions of the POA showed an increase in mean spine density at the age of vaginal opening. The increase in dendritic spine density in the posterior inferior quadrant of POA occurred prior to vaginal opening. The entire posterior POA maintained peak levels of spine numbers through 55 days of age, but then exhibited a dramatic drop at 75 days. Thus, modification of the neuropil may well continue at least to this age. The increases in spine densities at vaginal opening for the superior POA quadrants, followed by relatively stable values, appear to implicate these areas in the timing of the onset of puberty.     

Responses of preoptic thermosensitive neurons to medial forebrain bundle stimulation

HORI, T., T. KIYOHARA, T. NAKASHIMA AND M. SHIBATA. Responses of preoptic thermosensitive neurons tomedial forebrain bundle stimulation. BRAIN RES. BULL. 8(6) 667–675, 1982.—Single-unit responses of neurons in the preoptic and anterior hypothalamus (PO/AH) to local thermal stimulation and electrical stimulation of the medial forebrain bundle (MFB) were studied in urethane-anesthetized male rats. In a total of 286 units (112 warm-units, 37 cold-units and 137 thermally insensitive units), 109 units (49 warm-units, 13 cold-units and 47 thermally insensitive units) responded to single pulse stimulation of MFB. The units initially inhibited by MFB stimulation corresponded to 64.2% (70 of 109), the units with facilitatory responses were 27.5% (30 of 109) and the antidromically activated units were 8.3% (9 of 109). High incidence of inhibition by noradrenaline (NA) applied iontophoretically was observed in the neurons inhibited by the MFB stimulation. Iontophoretic application of dichloroisoproterenol to 2 warm-units blocked both the NA-induced inhibition and the MFB-induced inhibition. These ascending and descending connections of the MFB with PO/AH thermosensitive neurons may be part of the neural circuits responsible for thermoregulation.     

A newly identified mouse hypothalamic area having bidirectional neural connections with the lateral septum: the perifornical area of the anterior hypothalamus rich in chondroitin sulfate proteoglycans

While previous studies and brain atlases divide the hypothalamus into many nuclei and areas, uncharacterised regions remain. Here, we report a new region in the mouse anterior hypothalamus (AH), a triangular‐shaped perifornical area of the anterior hypothalamus (PeFAH) between the paraventricular hypothalamic nucleus and fornix, that abundantly expresses chondroitin sulfate proteoglycans (CSPGs). The PeFAH strongly stained with markers for chondroitin sulfate/CSPGs such as Wisteria floribunda agglutinin and antibodies against aggrecan and chondroitin 6 sulfate. Nissl‐stained sections of the PeFAH clearly distinguished it as a region of comparatively low density compared to neighboring regions, the paraventricular nucleus and central division of the anterior hypothalamic area. Immunohistochemical and DNA microarray analyses suggested that PeFAH contains sparsely distributed calretinin‐positive neurons and a compact cluster of enkephalinergic neurons. Neuronal tract tracing revealed that both enkephalin‐ and calretinin‐positive neurons project to the lateral septum (LS), while the PeFAH receives input from calbindin‐positive LS neurons. These results suggest bidirectional connections between the PeFAH and LS. Considering neuronal subtype and projection, part of PeFAH that includes a cluster of enkephalinergic neurons is similar to the rat perifornical nucleus and guinea pig magnocellular dorsal nucleus. Finally, we examined c‐Fos expression after several types of stimuli and found that PeFAH neuronal activity was increased by psychological but not homeostatic stressors. These findings suggest that the PeFAH is a source of enkephalin peptides in the LS and indicate that bidirectional neural connections between these regions may participate in controlling responses to psychological stressors.     

Extracellular serotonin variations during vigilance states in the preoptic area of rats: a microdialysis study

Numerous studies have shown that serotonergic transmission decreases from waking (W) to slow wave sleep (SWS) to paradoxical sleep (PS), suggesting an active role of serotonin (5-HT) in W but not in sleep. Conversely, the inhibition of 5-HT activity produces insomnia. This insomnia can be reversed by injections of 5-hydroxytryptophan in the preoptic area (POA), suggesting that 5-HT is necessary in this cerebral structure for sleep. Using microdialysis, we studied, 5-HT variations in the POA of rats in relation to vigilance states. 5-HT levels were higher during W than during during SWS and PS. 5-HT increased just before the rats fell asleep and then decreased during sleep. A decreased 5-HT transmission was also observed from SWS to PS. These data document a positive correlation between 5-HT levels in POA and wakefulness. Moreover, these observations are in favour of a permissive role of 5-HT in the POA during PS. A comparison between the POA and the prefrontal cortex in the sleep–wake cycle is discussed.     

Co-localization of enkephalin and TRH in perifornical neurons of the rat hypothalamus that project to the lateral septum

Neurons of the lateral septal nucleus are surrounded by terminals immunoreactive for thyrotropin-releasing hormone (TRH) and enkephalin (Enk). Retrograde labeling from the lateral septum in combination with immunocytochemical analyses for Enk and TRH in colchicine-treated rats has revealed that nearly all Enk- (and TRH-) containing perifornical neurons project to the lateral septum. Immunostaining of adjacent, thin paraffin sections for either TRH or Enk and double staining of thick vibratome sections for the two peptides have shown that TRH and Enk immunoreactivities co-exist within the same neurons in the perifornical region of the hypothalamus.     

Fos expression induced by warming the preoptic area in rats

The preoptic area (POA) occupies a crucial position among the structures participating in thermoregulation, but we know little about its efferent projections for controlling various effector responses. In this study, we used an immunohistochemical analysis of Fos expression during local warming of the preoptic area. To avoid the effects of anesthesia or stress, which are known to elicit Fos induction in various brain regions, we used a novel thermode specifically designed for chronic warming of discrete brain structures in freely moving rats. At an ambient temperature of 22 degrees C, local POA warming increased Fos immunoreactivity in the supraoptic nucleus (SON) and the periaqueductal gray matter (PAG). Exposure of animals to an ambient temperature of 5 degrees C induced Fos immunoreactivity in the magnocellular paraventricular nucleus (mPVN) and the dorsomedial region of the hypothalamus (DMH). Concurrent warming of the POA suppressed Fos expression in these areas. These findings suggest that thermal information from the preoptic area sends excitatory signals to the SON and the PAG, and inhibitory signals to the mPVN and the DMH.     

Estrogen-receptor immunoreactivity in hamster brain: preoptic area, hypothalamus and amygdala

The distribution of estrogen-receptor containing cells in the preoptic area, hypothalamus and amygdala of female Syrian hamster brain was studied by immunocytochemical methods. Dense populations of estrogen-receptor immunoreactive (ER-IR) cells were found in the medial preoptic area, the bed nucleus of the stria terminalis, amygdala, ventral and lateral parts of the hypothalamus, and the arcuate nucleus. Injection of estradiol caused a decrease in estrogen-receptor immunoreactivity (ERIR) containing cells within one hour, a decrease that may reflect a change in the ability of the occupied estrogen receptor to bind the particular antibody (H222) used rather than down-regulation of the estrogen receptor. Our findings on the distribution of estrogen-receptor containing cells in these areas using an immunocytochemical technique are consistent with and extend the findings of others using autoradiographic and in vitro binding techniques to study estrogen receptor distribution in hamster brain.     

Behavioural and neuronal activation after microinjections of AMPA and NMDA into the perifornical lateral hypothalamus in rats

The perifornical lateral hypothalamic area (PeFLH), which houses orexin/hypocretin (OX) neurons, is thought to play an important role in arousal, feeding, and locomotor activity. The present study examined behavioural effects of activating PeFLH neurons with microinjections of ionotropic glutamate receptor agonists. Three separate unilateral microinjections of either (1) AMPA (1 and 2 mM in 0.1 μL artificial cerebrospinal fluid, ACSF) and ACSF, or (2) NMDA (1 and 10 mM in 0.1 μL ACSF), and ACSF were made into the PeFLH of adult male rats. Following each injection, the rats were placed into an open field for behavioural scoring for 45 min. Rats were perfused after the third injection for immunohistochemistry for c-Fos and OX to assess the level of activation of OX neurons. Behavioural analyses showed that, as compared to ACSF conditions, AMPA injections produced a dose-dependent increase in locomotion and rearing that persisted throughout the 45 min recording period, and an increase in drinking. Injection of NMDA at 10 mM, but not 1 mM, induced a transient increase in locomotion and an increase in feeding. Histological analyses showed that while both agonists increased the number of neurons immunoreactive for c-Fos in the PeFLH, only AMPA increased the number of neurons immunoreactive for both c-Fos and OX. There were positive correlations between the number of c-Fos/OX-immunoreactive neurons and the amounts of locomotion, rearing, and drinking. These results support the role of ionotropic glutamate receptors on OX and other neurons in the PeFLH in the regulation of locomotor and ingestive behaviours.     

Changes in body temperature and thermosensitivity of preoptic and septal neurons during hippocampal stimulation

The effects of electrical stimulation of the ventral subiculum (VSB) of hippocampal formation on the thermosensitivity of neurons in the preoptic area and the septum (PO/SP) were studied in the urethane-anesthetized rat. The local thermosensitivity of sixteen of 28 PO/SP thermosensitive neurons (25 warm-units and 3 cold-units) was reduced or lost during low frequency (4.8 Hz) stimulation of VSB. The extent of reduction was positively correlated to the magnitude of inhibition by the VSB stimulation. Thermosensitivity did not change in 11 of the remaining 12 warm-units and increased slightly in one warm-unit during VSB stimulation. In the conscious rat, bilateral stimulation (5 Hz) of VSB for 10 min caused a rise in rectal temperature of 0.28-0.87 degrees C in a warm environment (30-33 degrees C) and a fall of 0.18-0.57 degrees C in a cold (9.4-9.8 degrees C) environment. The results suggest that the predominantly inhibitory input from VSB decreases the thermosensitivity of PO/SP warm- and cold-sensitive neurons and thereby reduces the ability of thermoregulation.     

Analysis of brainstem A1 and A2 noradrenergic inputs to the preoptic area using microdialysis in the rat

Noradrenergic inputs to the preoptic area (POA) are involved in regulating a variety of homeostatic functions. However, the accurate measurement of endogenous noradrenaline (NA) release in the POA has been difficult to achieve and consequently little has been done to characterise the different noradrenergic pathways. By combining the technique of intracranial microdialysis with tissue pre-loading of [³H]NA we have developed a sensitive index of NA release in the POA [8]. Using this method we have now examined and compared the effects of electrical stimulation of the brainstem A1 and A2 cell groups on NA release in the POA. Anaesthetized proestrus rats were implanted with microdialysis probes either unilaterally or bilaterally in the POA and stimulating electrodes positioned in either the A1 or A2 regions. Electrical stimulation (10 Hz, 10s on/off for 20 min) of the A1 region resulted in repeatable, calcium-dependent increases in radioactivity outflow from the ipsilateral POA(P < 0.01). A1-evoked release was twice as large as that observed after equivalent 10 Hz electrical stimulation of the A2 region(P < 0.05). In experiments using bilateral POA microdialysis and A1 stimulation, a significant increase in release from the contralateral POA, amounting to approximately 80% of that observed in the ipsilateral POA, was observed. Experiments involving the blockade of A1-stimulated release in the ipsilateral POA by perfusion with a calcium-free medium demonstrated that increases in radioactivity measured in the contralateral POA were not originating from the ipsilateral POA. These results provide a functional evaluation of the A1 and A2 noradrenergic inputs to the POA and provide evidence that (i) electrical activation of the A2 cell group increases NA release in the POA, although stimulated release is less than that resulting from equivalent A1 stimulation, and that (ii) stimulation of the A1 group alters NA release bilaterally in the POA.     

Carbamazepine Effects on Preoptic GABA Release and Pituitary Luteinizing Hormone Secretion in Rats

In vivo effects of carbamazepine (CBZ) on the neuroendocrine preopticopituitary feedback system were studied by local application of CBZ through a push—pull cannula into the preoptic area and measurement of local effects on γ-aminobutyric acid (GABA) and distant effects on a subsequent biologic response: luteinizing hormone (LH). Perfusion with 8 and 12 μg CBZ/ml cerebro-spinal fluid caused a reduction in preoptic GABA release and concomitant suppression of plasma LH levels. These results suggest a GABA component to the mechanism(s) of action of CBZ: (a) CBZ reduces extracellular available GABA concentration, and (b) owing to the known inhibitory role of preoptic GABA in pituitary LH secretion, an increase of postsynaptic GABAergic transmission by CBZ itself could be inferred.     

Reticulospinal neurons inactivated by warming of the preoptic area and anterior hypothalamus of rabbits

To identify the premotor neurons for vasoconstrictors of the skin, activities of reticulospinal neurons in the rostroventral medulla, the ear sympathetic nerve (ESNA) and the renal sympathetic nerve (RSNA) were recorded in anesthetized and immobilized Japanese White or New Zealand White rabbits. Two groups of neurons were identified according to their responses to thermal stimulation of the preoptic area and the anterior hypothalamus (POAH) and to electrical stimulation of baroreceptor afferents, the aortic nerve (AN). Neurons (Type I neurons, n = 21) whose activity was inhibited by warm stimulation of the POAH but not inhibited by the AN stimulation were located in sites medial to the rostral ventrolateral medulla (RVLM). The other neurons (Type II neurons, n = 20) whose activity was not inhibited by warm stimulation of the POAH but inhibited by the AN stimulation were located in the RVLM. Because the time course of the inhibitory response of Type I neurons to warm stimulation of the POAH was very similar to that of the inhibitory response of the ESNA and activities of these neurons and the ESNA were not inhibited by the stimulation of the AN, it was suggested the Type I neurons might participate in regulation of activity of the vasoconstrictors of the ear skin. The Type II neurons are considered to be the barosensitive RVLM neurons that regulate systemic arterial pressure by controlling the activity of visceral or muscular sympathetic vasoconstrictors or cardiac sympathetic fibers.     

A comparison of the effects of electrical stimulation of the lateral and ventromedial hypothalamus on the activity of neurons in ventral tegmental area and substantia nigra

Extracellular unit recordings were obtained from neurons in the ventral tegmental area (VTA), the substantia nigra, zona compacta (SNC) and zona reticulata (SNR) of adult female albino rats anaesthetized with urethane and chloral hydrate. Neurons were divided into two types based on their electrophysiological characteristics; Type I neurons had long duration action potentials (>2.6 msec) and slow discharge rates and Type II neurons had shorter duration action potentials and a wider range of discharge rates. Both types of neurons were found in the VTA and SNC, but there were only Type II neurons in the SNR. The effects of single pulse stimuli delivered to the ipsilateral ventromedial (VMH) or lateral (LH) hypothalamic areas on activities of the two types of neurons were investigated. Only a small portion of neurons in the VTA and SNC responded to VMH stimulation, but in contrast a majority of the two types of neurons in the VTA and SNC responded to LH stimulation. Most neurons in the SNR did not respond to VMH or to LH stimulation. Type II neurons in the VTA and SNC were predominantly suppressed by LH stimulation with short onset latencies (<6 msec), indicating the possibility of monosynaptic mediation. However Type I neurons in the VTA and SNC were activated and suppressed and the onset latencies of these responses were relatively longer. The high proportion of neurons of VTA and SNC responding to electrical stimulation of LH is consistent with anatomical evidence. Suppression and activation of Type I neurons in VTA and SNC suggest that the LH exerts modulatory influences on these neurons of the midbrain.     

Effects of endotoxin and sodium salicylate on the preoptic thermosensitive neurons in tissue slices

Effects of E. coli endotoxin and sodium salicylate (Sal) on single-unit activity of thermosensitive neurons recorded in slices of preoptic and anterior hypothalamic area (PO/AH) were studied in vitro. Perfusion of endotoxin-containing Krebs-Ringer's solution or local application of endotoxin in the immediate vicinity of recording neurons decreased and increased the firing rate of 31 of 34 warm-sensitive neurons and a cold-sensitive neuron, but had no effect on the majority of thermally insensitive neurons. In about half of warm-sensitive neurons the inhibitory response to endotoxin was preceded by a transient increase in firing rate. The pyrogen-induced decrease in firing rate in warm-sensitive neurons was reversibly blocked or attenuated by local application of Sal in a dose-dependent manner. The results are consistent with the view that pyrogen and Sal act in the PO/AH to produce fever and antipyresis, respectively, by appropriately offsetting the activity of thermosensitive neurons.     

Fos expression within regions of the preoptic area, hypothalamus and brainstem during pregnancy and parturition

Vaginocervical stimulation, that occurs during mating or with the birth of pups, is believed to induce specific sexual and maternal behaviours in the rat as well as stimulating a number of neuroendocrine responses including the secretion of oxytocin, prolactin and luteinizing hormone. Since the medial preoptic area has been implicated in the induction of maternal behaviour, the expression of the immediate-early gene product Fos was compared between non-pregnant, late pregnant and parturient rats. Although no difference was detected in the number of Fos-positive neuronal profiles in the preoptic area of non-pregnant and late-pregnant rats, a large increase was observed in the medial preoptic nucleus and the anteroventral periventricular region, as well as in the hypothalamic supraoptic nucleus, of parturient rats. Double labelling for Fos and tyrosine hydroxylase immunoreactivity in the brainstem of parturient rats showed the activation of catecholaminergic neurons in both the nucleus of the tractus solitarius and in the ventrolateral medulla that may form part of the afferent pathway from the uterus and cervix to the preoptic area and hypothalamus.