Trends in ambulance dispatches related to heat illness from 2010 to 2019: An ecological study.

Heatstroke is a serious heat-related illness that can even cause death. Heat alert systems play an important role in reducing the number of patients experiencing heat illness, as they encourage preventive actions such as the use of air conditioning, hydration, or other strategies. However, to date, the Japanese hazard classification has not considered seasonal and regional variations, despite clear differences in meteorological conditions across different regions in Japan. Moreover, several studies have reported a difference in thermoregulation between older and younger adults, implying that the hazard classification should also consider age differences. This study examined the relationship between the number of ambulance dispatches related to heat illness (ADRHI) and the Japanese heat hazard classification from 2010 to 2019, focusing on monthly and regional differences. Data from 47 prefectures during the 10-year period were collected and analyzed. ADRHI and wet bulb globe temperature (WBGT) data were collected from Japan's Ministry of Internal Affairs and Communications and the Ministry of the Environment Heat Illness Prevention Information website, respectively. The findings showed a significant relationship between ADRHI and WBGTmax (p < 0.05, r = 0.74). ADRHI per 100,000 people showed significant differences across months. The post hoc test detected the first steep increase in ADRHI at a WBGTmax of 23°C than at 22°C in June, and at a WBGTmax of 26°C, 27°C, and 25°C in July, August, and September, respectively. Moreover, the first significant increase in ADRHI per 100,000 people at WBGTmax differed across each region, at a WBGTmax of 24°C in Hokkaido-Tohoku, 25°C in Kanto, Kansai, and Chugoku, 26°C in Chubu, 27°C in Shikoku, and 28°C in Kyushu-Okinawa. Further, Poisson regression analysis revealed that the relative risks differed across each region and month. These results imply that the hazard classification should be adjusted according to region and month in Japan.

Effect of short- and long-term heat exposure on brain monoamines and emotional behavior in mice and rats.

Heat exposure affects several physiological, neuronal, and emotional functions. Notably, monoaminergic neurotransmitters in the brain such as noradrenaline, dopamine, and serotonin, which regulate several basic physiological functions, such as thermoregulation, food intake, and energy balance, are affected by heat exposure and heat acclimation. Furthermore, cognition and emotional states are also affected by heat exposure and changes in brain monoamine levels. Short-term heat exposure has been reported to increase anxiety in some behavioral tests. In contrast, there is a possibility that long-term heat exposure decreases anxiety due to heat acclimation. These changes might be due to adaptation of the core body temperature and/or brain monoamine levels by heat exposure. In this review, we first outline the changes in brain monoamine levels and thereafter focus on changes in emotional behavior due to heat exposure and heat acclimation. Finally, we describe the relationships between emotional behavior and brain monoamine levels during heat acclimation.

Comparison of neurotransmitter levels, physiological conditions, and emotional behavior between isolation-housed rats with group-housed rats.

Brain monoaminergic neurotransmitters, such as dopamine (DA), serotonin (5-HT), and noradrenaline (NA), play crucial roles in neuronal and physiological functions, including social behaviors. Isolation housing may induce behavioral and neurochemical abnormalities in rats, although its influence on neurotransmitter levels remains obscure. This study investigated the influence of isolation- or group-housing on core body temperature (Tcore ), locomotor activity (ACT), emotional behavior, and neurotransmitter levels in male Wistar rats. Behavioral changes were monitored using the open field test (OFT) and social interaction test (SIT). After 4 weeks, brain tissues were collected to quantify 5-HT, DA, and NA concentrations. Body weight and basal Tcore during both the light and dark phase were higher in isolation-housed than in group-housed rats, although no significant difference was seen in ACT. No significant differences were observed during the OFT. Isolation-housed rats showed increased line crossing and decreased social behavior during the SIT. Isolation-housed rats exhibited decreased levels of 5-HT in the caudate putamen and amygdala, and elevated and decreased NA levels in the paraventricular hypothalamic nucleus and hippocampus, respectively. However, DA levels were unaffected. Thus, housing environments may affect brain areas that regulate various neuronal and physiological functions, such as memory, stress responses, and emotional behavior.

Difference in the brain serotonin and its metabolite level and anxiety-like behavior between forced and voluntary exercise conditions in rats.

Physical exercise is beneficial to both physical and mental health, though it is unclear whether voluntary and forced exercise have the same effects. We investigated the effects of chronic forced and voluntary wheel running on brain levels of serotonin (5-HT), its metabolite 5-hydroxyindoleacetic acid (5-HIAA), and anxiety-like behavioral change in rats. Forty-eight rats were randomly assigned to standard cages (sedentary control: SC); voluntary exercise (free running on a wheel, V-EX); voluntary limited exercise (wheel available only 1 h per day, VL-EX); and forced exercise (running on a motorized wheel, F-EX). After 4 weeks, rats either underwent the open field test (OFT) or their 5-HT and 5-HIAA levels were measured in the major serotonergic neural cell bodies and projection areas. 5-HT and 5-HIAA levels in the dorsal and median raphe nuclei were increased in the V-EX, but not in the VL-EX and F-EX groups, compared with the SC group. In the paraventricular hypothalamic nucleus and caudate putamen, only 5-HT levels were increased in the V-EX group. Interestingly, in the amygdala, only 5-HIAA levels were significantly increased in the V-EX group. Conversely, we found that F-EX rats showed no significant 5-HT changes and increased anxiety-like behavior. VL-EX did not have significant beneficial effects on any of the experimental parameters. These data suggest that only unlimited voluntary exercise stimulates the serotonergic system and suppresses anxiety-like behavior.

Effect of heat acclimation on anxiety-like behavior of rats in an open field.

To mitigate the impacts of heat exposure, animals can take some actions to maintain their core body temperature, such as heat acclimation; however, the effect of heat acclimation on anxiety-like behavior in an open field is still not understood. The purpose of this study was to examine the anxiety-like behavior of heat acclimated rats in a temperate or heated open field. After being raised in a 23 °C environment for one week, male Wistar rats were exposed to a heated environment (32 °C) for 3 h (3H), 14 days (14D), or 28 days (28D), with free access to food and water, and compared with rats reared in a temperate environment (23 °C; Cont). After heat exposure, behavioral changes were monitored using an open field test (OFT) in a heated (32 °C) or temperate environment (23 °C). Compared with those in the Cont group, the body weights of rats in the 14D and 28D groups were lower. The OFT in the heated environment showed that grooming time was longer in 3H and 14D rats. In the temperate environment, grooming time was longer in all the heated groups. Rats from the 3H and 28D groups spent longer time in the center square when tested in the temperate environment. Rearing activity increased in 28D rats in the temperate environment, while the number of line crossings did not differ significantly between the heated groups and the two open fields. These results suggest that heat acclimation affected not only the physiological index such as core body temperature but also the anxiety-like behavior, mainly in the temperate open field. These changes might be beneficial when rats are faced with an open field.

Changes in thermoregulation and monoamine release in freely moving rats during cold exposure and inhibition of the ventromedial, dorsomedial, or posterior hypothalamus.

The hypothalamus is critical for regulating thermogenesis, but the role of monoamines in specific hypothalamic subregions in thermogenesis is not thoroughly established. The purpose of this study was to confirm changes of body temperature (T b) and thermoregulatory parameters upon inhibition of neural activity in hypothalamic subregions in freely moving rats. In addition, the pattern of monoamine release in these nuclei was measured during active thermoregulation using microdialysis. Tetrodotoxin (TTX) was perfused into the ventromedial hypothalamus (VMH), dorsomedial hypothalamus (DMH), or posterior hypothalamus (PH) at two different ambient temperatures (5 or 23 °C). Using telemetry, we continuously measured the T b and the heart rate (HR) as an index of heat production as well as locomotor activity (Act). Tail skin temperature (T tail) was also continuously measured as an index of heat loss. Although the perfusion of TTX into hypothalamic subregions had no effect on any of the measured thermoregulatory parameters at an ambient temperature of 23 °C, it induced significant T b decrease under cold conditions only when perfused into the DMH and the PH. In contrast, the HR decreased only after perfusion of TTX into the PH during cold conditions, while the T tail and Act remained unchanged. Serotonin (5-HT) in the DMH and dopamine (DA) metabolite 3,4-Dihydroxyphenylacetic acid in the PH, but not noradrenaline, increased significantly during exposure to cold temperatures. Our results indicate that the DMH and the PH, but not the VMH, are particularly involved in heat production under cold conditions. In addition, 5-HT in the DMH and DA in the PH may be involved in thermogenesis.

Involvement of serotonin in the ventral tegmental area in thermoregulation of freely moving rats.

We have recently reported that the serotonin (5-HT) projections from the midbrain's raphe nuclei that contains 5-HT cell bodies may play a role both in heat production and in heat loss. The purpose of the present study was to clarify the involvement of 5-HT in the ventral tegmental area (VTA), where 5-HT is suggested to participate in thermoregulation, using the combined methods of telemetry, microdialysis, and high performance liquid chromatography, with a special emphasis on regulation of the body temperature (Tb) in freely moving rats. First, we measured changes in Tb, tail skin temperature (Ttail; an index of heat loss), heart rate (HR; an index of heat production), locomotor activity (Act), and levels of extracellular monoamines in the VTA during cold (5°C) or heat (35°C) exposure. Subsequently, we perfused citalopram (5-HT re-uptake inhibitor) into the VTA and measured the thermoregulatory parameters and monoamines release. Although Tb, Ttail, and HR changed during both exposures, significant changes in extracellular level of 5-HT (138.7±12.7% baseline, p<0.01), but not dopamine (DA) or noradrenaline (NA) were noted in the VTA only during heat exposure. In addition, perfusion of citalopram into the VTA increased extracellular 5-HT levels (221.0±52.2% baseline, p<0.01), but not DA or NA, while Tb decreased from 37.4±0.1°C to 36.8±0.2°C (p<0.001),Ttail increased from 26.3±0.4°C to 28.4±0.4°C (p<0.001), and HR and Act remained unchanged. Our results suggest that the VTA is a key area for thermoregulation, and 5-HT, but not DA or NA, modulates the heat loss system through action in the VTA.

Exercise increases mTOR signaling in brain regions involved in cognition and emotional behavior.

Exercise can enhance learning and memory and produce resistance against stress-related psychiatric disorders such as depression and anxiety. In rats, these beneficial effects of exercise occur regardless of exercise controllability: both voluntary and forced wheel running produce stress-protective effects. The mechanisms underlying these beneficial effects of exercise remain unknown. The mammalian target of rapamycin (mTOR) is a translation regulator important for cell growth, proliferation, and survival. mTOR has been implicated in enhancing learning and memory as well as antidepressant effects. Moreover, mTOR is sensitive to exercise signals such as metabolic factors. The effects of exercise on mTOR signaling, however, remain unknown. The goal of the present study was to test the hypothesis that exercise, regardless of controllability, increases levels of phosphorylated mTOR (p-mTOR) in brain regions important for learning and emotional behavior. Rats were exposed to 6 weeks of either sedentary (locked wheel), voluntary, or forced wheel running conditions. At 6 weeks, rats were sacrificed during peak running and levels of p-mTOR were measured using immunohistochemistry. Overall, both voluntary and forced exercise increased p-mTOR-positive neurons in the medial prefrontal cortex, striatum, hippocampus, hypothalamus, and amygdala compared to locked wheel controls. Exercise, regardless of controllability, also increased numbers of p-mTOR-positive glia in the striatum, hippocampus, and amygdala. For both neurons and glia, the largest increase in p-mTOR positive cells was observed after voluntary running, with forced exercise causing a more modest increase. Interestingly, voluntary exercise preferentially increased p-mTOR in astrocytes (GFAP+), while forced running increased p-mTOR in microglia (CD11+) in the inferior dentate gyrus. Results suggest that mTOR signaling is sensitive to exercise, but subtle differences exist depending on exercise controllability. Increases in mTOR signaling could contribute to the beneficial effects of exercise on cognitive function and mental health.

Possible mechanisms of hypothermia after inhibition of the median or dorsal raphe nucleus of freely moving rats.

We previously reported that tetrodotoxin (TTX) perfusion into the median raphe nucleus (MRN), which contains the cell bodies of serotonin (5-HT) neurons, induced a considerable body temperature reduction under normal and low ambient temperatures (23 and 5°C, respectively) in freely moving rats but showed no such effect under high ambient temperature (35°C). In the present study, we aimed to determine the mechanism(s) of body temperature reduction after TTX perfusion into the MRN by measuring tail skin temperature (an index of heat loss), heart rate (an index of heat production), and locomotor activity (Act) under normal ambient temperature (23°C). We performed similar experiments in the dorsal raphe nucleus (DRN), another area containing cell bodies of 5-HT neurons, to compare any functional differences with the MRN. TTX perfusion into the MRN or DRN induced significant hypothermia (from 37.4±0.2 to 33.7±0.4°C or from 37.4±0.1 to 34.5±0.4°C, respectively; P<0.001) with increased tail skin temperature (from 26.1±0.8 to 31.1±1.3°C or from 26.3±0.9 to 31.7±0.4°C, respectively; P<0.001), but no change in heart rate. However, TTX perfusion into the MRN or DRN differentially affected Act. TTX perfusion into the MRN induced hyperactivity (from 10.7±4.6 to 67.6±25.1 counts/min; P<0.01), whereas perfusion into the DRN induced immobility. Thus, the 5-HT projections from the MRN and DRN may play similar roles in thermoregulation, both in the heat production system and in the heat loss system, but their roles in the regulation of Act might be distinct and opposite.

Changes of brain monoamine levels and physiological indexes during heat acclimation in rats.

Brain monoamines, such as noradrenaline (NA), dopamine (DA), and serotonin (5-HT), regulate many important physiological functions including thermoregulation. The purpose of this study was to clarify changes in NA, DA, and 5-HT levels in several brain regions in response to heat acclimation while also recording body temperature (Tb), heart rate (HR), and locomotor activity (Act). Rats were exposed to a heated environment (32°C) for 3h (3H), 1 day (1D), 7 days, 14 days (14D), 21 days, or 28 days (28D). After heat exposure, each of the following brain regions were immediately extracted and homogenized: the caudate putamen (CPu), preoptic area (PO), dorsomedial hypothalamus (DMH), frontal cortex (FC), and hippocampus (Hip). NA, DA, and 5-HT levels in the extract were measured by high performance liquid chromatography. Although Tb increased immediately after heat exposure, it decreased about 14D later. HR was maintained at a low level throughout heat exposure, and Act tended to increase near the end of heat exposure. After 3H, we observed a marked increase in NA level in the CPu. Although this response vanished after 1D, the level increased again after 28D. DA level in the CPu decreased significantly from 1D to 28D. 5-HT level in the PO and DMH decreased from 1D to 14D. It returned to control levels after 28D with increment of DA level. 5-HT level in the FC decreased at the start of heat exposure, but recovered after 28D; a time point at which DA level also increased. Monoamine levels in the Hip were unchanged after early heat exposure, but both 5-HT and DA levels increased after 28D. These results provide definitive evidence of changes in monoamines in individual brain regions involved in thermoregulation and behavioral, cognitive, and memory function during both acute and chronic heat exposure.

Influence of circadian disruption on neurotransmitter levels, physiological indexes, and behaviour in rats.

Brain monoamines - such as noradrenaline (NA), dopamine (DA) and serotonin (5-HT) - regulate several important physiological functions, including the circadian rhythm. The purpose of this study was to examine changes in NA, DA and 5-HT levels in various brain regions and their effect on core body temperature (Tc), heart rate (HR) and locomotor activity (Act) in rats following exposure to an artificial light/dark (LD) cycle. For this, male Wistar rats were housed at an ambient temperature (Ta) of 23 °C and 50% relative humidity with free access to food and water. Rats were exposed to either natural (12 h:12 h) or artificial (6 h:6 h) LD cycles for 1 month, after which each brain region was immediately extracted and homogenized to quantify the amounts of NA, DA and 5-HT by high-performance liquid chromatography. Behavioural changes were also monitored by the ambulatory activity test (AAT). Notably, we found that artificial LD cycles disrupted the physiological circadian rhythms of Tc, HR and Act. Although the 5-HT levels of rats with a disrupted circadian rhythm decreased in cell bodies (dorsal and median raphe nuclei) and projection areas (frontal cortex, caudate putamen, preoptic area and suprachiasmatic nucleus) relative to the control group, NA levels increased both in the cell body (locus coeruleus) and projection area (paraventricular hypothalamus). No significant changes were found with respect to DA. Moreover, circadian rhythm-disrupted rats also showed anxious behaviours in AAT. Collectively, the results of this study suggest that the serotonergic and noradrenergic systems, but not the dopaminergic system, are affected by artificial LD cycles in brain regions that control several neural and physiological functions, including the regulation of physiological circadian rhythms, stress responses and behaviour.

Source analysis of stimulus-preceding negativity constrained by functional magnetic resonance imaging.

The stimulus-preceding negativity (SPN) is an event-related potential (ERP) reflecting anticipation. The anterior insular cortex is assumed to be one of the physiological sources of the SPN. However, the precise neural substrates of the SPN have yet to be confirmed. We therefore performed separate functional magnetic resonance imaging (fMRI) and ERP studies using the same time estimation task, followed by fMRI-constrained ERP source analysis. Dipole locations were determined by the fMRI results, while the time courses of dipole activities were modeled by the ERP data. Analysis revealed that the right anterior insula was significantly activated before delivery of the feedback stimulus, whereas the left anterior insula was not, and that the SPN mainly arose from four groups of brain regions related to, respectively: (1) the salience network, (2) reward expectation, (3) perceptual anticipation, and (4) arousal. The results suggest that the SPN pertains to multiple brain functions with complex interactions.

Continuous monitoring of hypothalamic neurotransmitters and thermoregulatory responses in exercising rats.

The purpose of the present study was to investigate the relationship between thermoregulation and catecholamine release in the preoptic area and anterior hypothalamus (PO/AH) during incremental treadmill running in the rat. To this aim, we combined in vivo brain microdialysis, biotelemetry and metabolic measurements for continuous monitoring of core body temperature (T(core)), neurotransmitters and thermoregulatory responses. The animals were exercised for 1h at 23°C. Treadmill speed was increased every 20 min (10, 20 and 26 m min⁻¹). T(core), oxygen consumption (V˙O2, an index of heat production) and tail skin temperature (T(tail), an index of heat loss) were simultaneously measured. Brain microdialysis samples were collected every 10 min, and these samples were analyzed for noradrenaline (NA), dopamine (DA) and serotonin (5-HT). T(core) and V˙O(2)2 significantly increased during treadmill and were exercise intensity dependent. After an initial drop T(tail) increased significantly during exercise. Both NA and DA levels in the PO/AH increased significantly during exercise. There was no effect on serotonin release. T(core), V˙O2 and T(tail) were positively correlated with the levels of NA and DA. Our data suggest that thermoregulatory responses are dependent on the intensity of the exercise and that these responses are associated with changes in NA and DA release, but not in 5-HT release in the PO/AH.

Serotonin release in the preoptic area and anterior hypothalamus is not involved in thermoregulation during low-intensity exercise in a warm environment.

The aim of the present study was to assess the response of serotonin (5-hydroxytryptamine; 5-HT) in the preoptic area and anterior hypothalamus (PO/AH) to thermoregulation during exercise in a warm environment. In addition, it was investigated whether a rapid increase in extracellular 5-HT levels in the PO/AH modifies the thermoregulatory response under the same exercise conditions. Rats were made to run for 120 min at 10 m min(-1) on a treadmill at the ambient temperature of 30 degrees C. Body core temperature (T(core)) was monitored using a biotelemetry system, and tail skin temperature (T(tail)) was simultaneously measured as an index of heat loss response. Microdialysis in combination with HPLC was used to measure concentrations of monoamines in the PO/AH. Both T(core) and T(tail) increased during the first 20 min of exercise and remained stable until the end of the exercise period. Low-intensity exercise did not induce any changes in 5-HT release in the PO/AH, although the levels of norepinephrine and dopamine were increased. Moreover, increased extracellular 5-HT by local perfusion of 1 microM citalopram (selective 5-HT reuptake inhibitor; SSRI) in the PO/AH had no effect on the thermoregulatory response during acute low-intensity exercise in a warm environment. These results suggest that enhanced release of only 5-HT in the PO/AH may not intervene thermoregulation during exercise in a warm environment.

Effect of chronic cold exposure on noradrenergic modulation in the preoptic area of thermoregulation in freely moving rats.

For this study, we compared the thermoregulatory involvement of noradrenaline (NA) in the medial preoptic area (mPOA) of non-cold acclimated rats to that of cold-acclimated rats. We quantified the release of NA in the mPOA during 3 h cold (5 degrees C) exposure in room-temperature-acclimated rats (RA group, kept at 23 degrees C for 2 weeks) and cold-acclimated rats (CA group, kept at 5 degrees C for 2 weeks). We concurrently monitored the core body temperature (Tc), heart rate (HR), and tail skin temperature (Tt). Cold exposure significantly increased Tc and HR, and decreased Tt in both groups. However, the cold-induced increase of the extracellular NA levels in mPOA was observed only in the RA group: not in the CA group. To elucidate these different results in NA levels further, and to evaluate participation of the mPOA in thermoregulation in the cold, we measured Tc, HR, and Tt during perfusion of alpha-adrenoceptor antagonist phenoxybenzamine during cold exposure (5 degrees C). This pharmacological procedure induced marked hypothermia, with decreases in HR only in the RA group; no changes were observed in Tc or any thermoregulatory parameter in the CA group. These results suggest that NA in the mPOA modulates heat production in response to acute cold stress in the RA group. However, this thermoregulatory action of NA in the mPOA was attenuated in the CA group.

Influence of brain catecholamines on the development of fatigue in exercising rats in the heat.

The purpose of the present study was to identify the effects of an acute injection of a dual dopamine (DA)/noradrenaline (NA) reuptake inhibitor (bupropion) on exercise performance, thermoregulation and neurotransmitters in the preoptic area and anterior hypothalamus (PO/AH) of the rat during exercise in the heat. Body core temperature (T(core)), brain temperature (T(brain)) and tail skin temperature (T(tail)) were measured. A microdialysis probe was inserted in the PO/AH, and samples for measurement of extracellular DA, NA and serotonin (5-HT) levels were collected. Rats received either bupropion (17 mg kg(-1); hot-BUP) or saline (1 ml kg(-1); hot) 20 min before the start of exercise and ran at a speed of 26 m min(-1) until exhaustion in a warm environment (30 degrees C). Rats also ran until exhaustion in a cool environment (18 degrees C; cool). Running time to exhaustion was significantly influenced by the ambient temperature, and it was increased by bupropion in the heat (cool, 143.6 +/- 21 min; hot, 65.8 +/- 13 min; hot-BUP, 86.3 +/- 7.2 min). T(core) and T(brain) at exhaustion were significantly higher in the bupropion group compared to the cool and hot groups, respectively. T(tail) measured at exhaustion was not significantly different between the two hot conditions. Extracellular concentrations of DA and NA in the PO/AH increased during exercise, and was significantly higher in the bupropion than in cool and hot groups (P < 0.05). No differences were observed between groups for 5-HT levels. These results suggest that DA and NA in the PO/AH might be responsible for the increase in exercise performance and T(core) and T(brain) in the bupropion group in hyperthermia. Moreover, these results support previous findings in humans that acute bupropion ingestion increases T(core) during exercise in the heat, indicating the possibility of an important role for DA and NA in thermoregulation.

Influence on human sleep patterns of lowering and delaying the minimum core body temperature by slow changes in the thermal environment.

STUDY OBJECTIVES: We hypothesized that appropriate changes in thermal environment would enhance the quality of sleep. DESIGN/SETTING: Controlled laboratory study. PARTICIPANTS: Healthy young men (n = 7, mean age 26 years). INTERVENTIONS: Nocturnal sleep structures in semi-nude subjects were compared between a condition where an ambient temperature (Ta) of 29.5 degree C was maintained throughout the night (constant Ta), and a second condition (dynamic Ta) where Ta changed slowly within the thermoneutral range (from 27.5 C to 29.5 degree C). MEASUREMENTS AND RESULTS: Statistically significant (P < 0.05) results included a lower and a later occurrence of minimum core body temperature (Tc), and a longer duration of slow-wave (stages 3+4) sleep in dynamic versus constant T. However, total sleep time, sleep efficiency, the total durations of light (stages 1+2) and rapid eye movement sleep, and the latencies to sleep onset, slow-wave sleep, and rapid eye movement sleep did not differ between conditions. CONCLUSIONS: Lowering the minimum and delaying the nadir of nocturnal Tc increases slow-wave sleep (probably by an increase of dry heat loss); use of this tactic might improve the overall quality of sleep.

Changes of body temperature and thermoregulatory responses of freely moving rats during GABAergic pharmacological stimulation to the preoptic area and anterior hypothalamus in several ambient temperatures.

Action of gamma-aminobutyric acid (GABA) in the preoptic area and anterior hypothalamus (PO/AH) has been implicated to regulate body temperature (T(b)). However, its precise role in thermoregulation remains unclear. Moreover, little is known about its release pattern in the PO/AH during active thermoregulation. Using microdialysis and telemetry techniques, we measured several parameters related to thermoregulation of freely moving rats during pharmacological stimulation of GABA in normal (23 degrees C), cold (5 degrees C), and hot (35 degrees C) ambient temperatures. We also measured extracellular GABA levels in the PO/AH during cold (5 degrees C) and heat (35 degrees C) exposure combined with microdialysis and high performance liquid chromatography (HPLC). Perfusion of GABA(A) agonist muscimol into the PO/AH increased T(b), which is associated with increased heart rate (HR), as an index of heat production in all ambient temperatures. Although tail skin temperature (T(tail)) as an index of heat loss increased only under normal ambient temperatures, its response was relatively delayed in comparison with HR and T(b), suggesting that the increase in T(tail) was a secondary response to increased HR and T(b). Locomotor activity also increased in all ambient temperatures, but its response was not extraordinary. Interestingly, thermoregulatory responses were different after perfusion of GABA(A) antagonist bicuculline at each ambient temperature. In normal ambient temperature conditions, perfusion of bicuculline had no effect on any parameter. However, under cold ambient temperature, the procedure induced significant hypothermia concomitant with a decrease in HR in spite of hyperactivity and increase of T(tail). It induced hyperthermia with the increase of HR but no additional change of T(tail) in hot ambient temperature conditions. Furthermore, the extracellular GABA level increased significantly during cold exposure. Its release was lower during heat exposure than in a normal environment. These results indicate that GABA in the PO/AH is an important neurotransmitter for disinhibition of heat production and inhibition of heat loss under cold ambient temperature. It is a neurotransmitter for inhibition of heat production under hot ambient temperature.

Inhibition of the preoptic area and anterior hypothalamus by tetrodotoxin alters thermoregulatory functions in exercising rats.

We have previously demonstrated a functional role of the preoptic area and anterior hypothalamus (PO/AH) in thermoregulation in freely moving rats at various temperature conditions by using microdialysis and biotelemetry methods. In the present study, we perfused tetrodotoxin (TTX) solution into the PO/AH to investigate whether this manipulation can modify thermoregulation in exercising rats. Male Wistar rats were trained for 3 wk by treadmill running. Body core temperature (Tb), heart rate (HR), and tail skin temperature (Ttail) were measured. Rats ran for 120 min at speed of 10 m/min, with TTX (5 microM) perfused into the left PO/AH during the last 60 min of exercise through a microdialysis probe (control, n=12; TTX, n=12). Tb, HR, and Ttail increased during the first 20 min of exercise. Thereafter, Tb, HR, and Ttail were stable in both groups. Perfusion of TTX into the PO/AH evoked an additional rise in Tb (control: 38.2 +/- 0.1 degrees C, TTX: 39.3 +/- 0.2 degrees C; P <0.001) with a significant decrease in Ttail (control: 31.2 +/- 0.5 degrees C, TTX: 28.3 +/- 0.7 degrees C; P <0.01) and a significant increase in HR (control: 425.2 +/- 12 beats/min, TTX: 502.1 +/- 13 beats/min; P <0.01). These results suggest that the TTX-induced hyperthermia was the result of both an impairment of heat loss and an elevation of heat production during exercise. We therefore propose the PO/AH as an important thermoregulatory site in the brain during exercise.

Changes of body temperature and extracellular serotonin level in the preoptic area and anterior hypothalamus after thermal or serotonergic pharmacological stimulation of freely moving rats.

Although many studies has been shown that serotonin (5-HT) in the preoptic area and anterior hypothalamus (PO/AH) is important for regulating body temperature (Tb), the exact role is not established yet due to conflicting results probably related to experimental techniques or conditions such as the use of anesthesia. The purpose of present study was to clarify the role of 5-HT in the PO/AH using the combined methods of telemetry, microdialysis and high performance liquid chromatography (HPLC), with a special emphasis on the regulation of Tb in freely moving rats. Firstly, we measured changes in Tb and levels of extracellular 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) in the PO/AH during cold (5 degrees C) and heat (35 degrees C) exposure. We also perfused fluoxetine (5-HT re-uptake inhibitor) and 8-hydroxy-2-(Di-n-propylamino)tetralin (8-OH-DPAT: 5-HT1A agonist) into the PO/AH. During both exposures, although Tb changed significantly, no significant changes were noted in extracellular levels of 5-HT and 5-HIAA in the PO/AH. In addition, although perfusion of fluoxetine or 8-OH-DPAT into the PO/AH increased or decreased extracellular 5-HT and 5-HIAA levels in the PO/AH respectively, but Tb did not change at all. Our results suggest that 5-HT in the PO/AH may not mediate acute changes in thermoregulation.