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.

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.

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.