Influence of the hypothalamus on the midbrain tonic inhibitory mechanism on metabolic heat production in rats

Influence of the hypothalamus on increased body temperature was examined in male rats. Body temperature was increased by removing the midbrain tonic inhibitory mechanism (TIM) on heat production from brown adipose tissue (BAT) by microinjections of a local anesthetic, procaine, into the midbrain. Procaine microinjections in unanesthetized rats increased rectal temperature that was followed by a strong tail skin temperature rise. Procaine microinjections in unanesthetized and decerebrated rats also increased rectal temperature but without skin temperature rise. These decerebrated animals fatally developed hyperthermia. In anesthetized rats, procaine microinjections increased temperature of the interscapular BAT (IBAT) higher with shorter onset for temperature rise than rectal temperature. Increased IBAT temperature by procaine microinjections in anesthetized rats was attenuated during hypothalamic warming, and enhanced during hypothalamic cooling when compared with that observed during thermoneutral hypothalamic temperature. These results suggest that the midbrain TIM is able to function in unanesthetized conscious rats, and that the integrity of the midbrain mechanism to tonically inhibit metabolic heat production does not require the presence of intact hypothalamus. These results also suggest that the hypothalamus modulates directly or indirectly IBAT heat production that was induced by removal of the midbrain TIM.     

Thermoresponsiveness of posterior hypothalamic (PH) neurons of rats to scrotal and abdominal thermal stimulation

The thermoresponsiveness of posterior hypothalamic (PH) neurons to localized, incremental thermal heating and cooling between 10–40°C of the abdomen or scrotum was determined in urethane anesthetized, male Sprague–Dawley rats whose core temperature was maintained at 37°C during testing. PH extracellular neuronal activity was recorded along with changes in gastrocnemius muscle EMG activity and temperature (T_ms, indicative of shivering thermogenesis) and intrascapular brown adipose tissue temperature (T_IBATs, indicative of non-shivering thermogenesis). Seventy-five PH neurons were recorded following both scrotal and abdominal trials of thermal stimulation. Nine percent of PH neurons were classified as warm responsive neurons (WRNs), 20% as cold responsive (CRNs), and 71% as temperature nonresponsive neurons (TNRNs), based on their thermal coefficients (TCs). Mean TC for warm PH neurons was significantly increased with scrotal warming between 30–40°C from the mean TC of the same PH WRNs following abdominal warming. Similarly, the thermal coefficient was increased (i.e., was more negative) for cold responsive PH neurons to scrotal cooling (20–10°C) as opposed to the TC of the same PH CRNs for abdominal cooling. No shivering thermogenesis (no change in temperature or EMG activity from gastrocnemius muscle) or non-shivering thermogenesis (no significant increase in IBAT temperatures) occurred with scrotal or abdominal cooling in these 21°C acclimatized rats. The results indicate that a small population of PH neurons are thermoresponsive to localized physiological changes in temperature of the scrotum and abdomen with greater thermoresponsiveness shown of both warm and cold PH neurons to scrotal vs. abdominal thermal stimulation.     

Response characteristics of specific thermoreceptive afferents innervating monkey facial skin and their relationship to human thermal sensitivity

The responses of thermoreceptive afferents to steady-state constant skin temperatures and temperature shifts were studied in rhesus monkeys by recording single fiber activity from fine dissected strands of the infraorbital nerve. Eighty-five warm and 134 cold fibers were identified by their specific sensitivity to warming and cooling of their receptive fields, which usually were restricted to single spots less than 1 mm in diameter. Warm fibers increased their firing rates with warming and decreased or ceased firing with cooling, while cold fibers showed an opposite behavior. Mean conduction velocity was 3.4 ± 1.7 m/s in warm fibers, and 9.0 ± 3.4 m/s in cold fibers.In response to constant skin temperatures in the innocuous temperature range below 43°C, warm fibers were active over the range of 30–42°C and showed maximum firing rates at 42°C. On the other hand, cold fibers were responsive to constant temperatures over the range of 20–42°C and their firing frequencies were maximal at 30°C and decreased as steady-state temperatures were raised or lowered. In addition, cold fibers exhibited periodic bursting activity when the temperature was below 35°C. The number of intraburst spikes in each burst increased as the constant temperature was     

Glutamate injection into the suprachiasmatic nucleus stimulates brown fat thermogenesis in the rat

Injection of glutamate (100 mM to 1 M, in 0.25 μl saline) into the hypothalamic suprachiasmatic nucleus (SCN) dose-dependently increased interscapular brown adipose tissue (IBAT) and core temperatures in the urethane-anaesthetized rat. This effect was more pronounced in rats tested during the light-off period than in animals tested during the light-on period. Prior injection of the local anaesthetic, procaine (5% in 0.5 μl saline), into the ipsilateral ventromedial hypothalamic nucleus (VMH) attenuated the increases in IBAT and core temperatures induced by intra-SCN glutamate. The VMH has previously been implicated in the central regulation of BAT thermogenesis; the present results suggest the pathway arising in the SCN exerts an excitatory influence on VMH neurons involved in the control of BAT function.     

N-Monomethyl-l-arginine co-injection attenuates the thermogenic and hyperthermic effects of E2 prostaglandin microinjection into the anterior hypothalamic preoptic area in rats

Prostaglandin E₂ (PGE₂) microinjection (25 ng, 250 nl) into the preoptic area of the anterior hypothalamus (POAH) stimulated heat production in brown adipose tissue (BAT) and increased core temperature in urethane-anesthetized rats. These thermogenic and hyperthermic effects were attenuated by co-injection of N^G-monomethyl-l-arginine (NMMA, 25 μg), a competitive inhibitor of nitric oxide (NO) production froml-arginine. Inclusion ofl-arginine (50 μg), though notd-arginine (50μg) reversed the inhibitory effect of NMMA (25μg) on intra-POAH PGE₂-induced increases in interscapular BAT (IBAT) and core temperatures. Intra-POAH injection of NMMA (25 μg) orl-arginine (50 μg) alone had no effect on IBAT and core temperatures. The results suggest that the effect on thermoregulation induced by action of PGE₂ in the POAH is modulated by a locall-arginine-dependent and NMMA-sensitive NO-generating system.     

Brown adipose tissue thermogenesis induced by low level electrical stimulation of hypothalamus in rats

Brown adipose tissue (BAT) is an energy dissipating form of adipose tissue implicated in non-shivering thermogenesis as well as diet-induced thermogenesis. In the present study, in vivo interscapular BAT (IBAT) temperature was recorded prior to and following low level electrical stimulation (a 30 sec train of 60 Hz, 100 microA 0.5 msec isolated pulses) of various hypothalamic regions in rats. Significant increases in IBAT temperature were observed after stimulation of the anterior, medial preoptic, paraventricular and dorsomedial hypothalamus but not after stimulation of either ventromedial or caudal hypothalamus. For positive sites, IBAT temperature typically increased at 3-4 minutes following stimulation, peaked at 7-8 minutes after stimulation and declined at 20 minutes after stimulation. Although alterations in diet-induced thermogenesis have been reported after ventromedial hypothalamic lesions, the increases in BAT temperature noted in the present study suggest that inhibitory fibers that course through the paraventricular hypothalamus may form part of the central nervous system control of brown adipose tissue thermogenesis induced by overfeeding.     

Injection of orexin A into the diagonal band of Broca induces sympathetic and hyperthermic reactions

This experiment tested the effect of an injection of orexin A into the diagonal band of Broca on the sympathetic activity and body temperature. Concentration of glycerol into white fat of lumbar region, firing rates of sympathetic nerves to interscapular brown adipose tissue (IBAT), IBAT and colonic temperatures, and heart rate were monitored in urethane-anesthetized male Sprague-Dawley rats for 30 min before and 150 min after injections of orexin A (0.4 and 0.7 nmol) into the diagonal band of Broca. The same variables were monitored in control rats with an injection of saline. The results show that orexin A increases glycerol concentration, sympathetic firing rate, IBAT and colonic temperatures, and heart rate. The saline injection did not induce any modification. These findings suggest that the diagonal band of Broca is a cerebral structure involved in the induction of the hyperthermia due to orexin A.     

Effects of variations in hippocampal slice preparation protocol on the electrophysiological stability, epileptogenicity and graded hypoxia responses of CA1 neurons

Evoked, extracellularly recorded field potentials and whole-cell current-clamp recordings were used to assay the effects of variations in dissection method and incubation temperature on the electrophysiology of CA1 neurons in hippocampal slices. Slices were cut with either a vibratome or a tissue chopper, and incubated at 28–30°C, room temperature (19–21°C), or in cool solution (13–15°C) which was allowed to passively warm to room temperature while the slices were incubating (‘cold-shock', CS). Although no effects of dissection method were observed, it was found that incubation temperature had profound effects on synaptically, but not non-synaptically evoked field potentials. Cold-shocked slices, cut with either a vibratome or a tissue chopper, exhibited epileptiform and spontaneously potentiating orthodromic field potentials. Slices incubated at warmer temperatures demonstrated responses that were larger in amplitude, more stable and much less epileptiform. In response to orthodromic stimulation, CS neurons fired more action potentials than did neurons in slices incubated at room temperature. Further, CS neurons generated smaller inhibitory postsynaptic potentials. Field potential changes resulting from graded hypoxia were not significantly affected by the temperature at which the slices were incubated. These data suggest that the electrophysiology of the hippocampal slice can be altered by the methods used to prepare the tissue. This finding may account for some of the discrepancies that exist between laboratories, and serves to underscore the importance of accurately reporting detailed protocols. Further, CS hippocampal tissue may represent a novel in vitro model of epileptiform activity.     

Inhibition of RVLM synaptic activation at peak hyperthermia reduces visceral sympathetic nerve discharge

Hyperthermia is an environmental stressor that produces marked increases in visceral sympathetic nerve discharge (SND) in young rats. The brainstem in rats contains the essential neural circuitry for mediating visceral sympathetic activation; however, specific brainstem sites involved remain virtually unknown. The rostral ventral lateral medulla (RVLM) is a key central nervous system region involved in the maintenance of basal SND and in mediating sympathetic nerve responses evoked from supraspinal sites. In the present study we tested the hypothesis that inhibition of RVLM synaptic activation at peak hyperthermia (internal body temperature, Tc, increased to 41.5 °C) would affect heating-induced visceral sympathetic activation. Experiments were completed in chloralose–urethane anesthetized, baroreceptor-intact and sinoaortic-denervated, 3–6 month-old Sprague–Dawley rats. Bilateral inhibition of RVLM synaptic activation produced by muscimol microinjections (400 and 800 pmol) at 41.5 °C resulted in immediate and significant reductions in peak heating-induced renal and splenic sympathoexcitation. Interruption of RVLM synaptic activation and axonal transmission by lidocaine microinjections (40 nmol) at 41.5 °C produced significant reductions in hyperthermia-induced sympathetic activation to similar levels produced by RVLM muscimol microinjections. The total amount of SND inhibited by RVLM muscimol and lidocaine microinjections was significantly more during hyperthermia (41.5 °C) than normothermia (38 °C). These findings demonstrate that maintenance of sympathetic activation at peak hyperthermia is dependent on the integrity of RVLM neural circuits.     

Intracerebroventricular injection of prostaglandin E(1) changes concentrations of biogenic amines in the posterior hypothalamus of the rat

Since the posterior hypothalamus (PH) plays a key role in the control of body temperature, the aim of this study was to evaluate the changes in adrenaline, noradrenaline and dopamine levels in the PH during the hyperthermia induced by prostaglandin E(1) (PGE(1)). The concentration of adrenaline, noradrenaline and dopamine in the PH, the firing rate of the sympathetic nerves innervating interscapular brown adipose tissue (IBAT), IBAT and colonic temperatures (T(IBAT) and T(C)) were monitored in 12 urethane-anaesthetized male Sprague-Dawley rats before and after an intracerebroventricular injection of 500 ng PGE(1) dissolved in 2 microl of 0.9% NaCl saline solution or only saline. The catecholamines were collected using a microdialysis probe and quantified by HPLC. The results showed that PGE(1) caused a significant increment in the concentration of adrenaline from 15. 83+/-2.69 to 34.95+/-3.9 ng ml(-1) and of dopamine from 35.15+/-4.48 to 55.68+/-6.21 ng ml(-1). A significant decrease in the level of noradrenaline from 18.75+/-2.05 to 8.56+/-2.26 ng ml(-1) was registered. The firing rate of sympathetic nerves to IBAT was increased from 100+/-0% to 204.83+/-15.22% by PGE(1). T(IBAT) and T(C) rose respectively from 36.91+/-0.15 degrees C to 38.88+/-0.29 degrees C, and from 36.7+/-0.15 degrees C to 38.13+/-0.36 degrees C after the injection of PGE(1). The changes in adrenaline and noradrenaline occurred during the first 20 min as did the changes in temperature and firing rate, while the change in dopamine was delayed until 21-60 min after the PGE(1) injection. No significant change of analyzed variables was found in the control rats. These findings suggest that these biogenic amines of the PH are involved in the control of the sympathetic and thermogenic changes induced by PGE(1).     

Hyperthermia complicates middle cerebral artery occlusion induced by an intraluminal filament

The present experiments were designed to study under what circumstances middle cerebral artery (MCA) occlusion by an intraluminal filament technique leads to hyperthermia and what the mechanisms are. We found that permanent MCA occlusion by this technique lead to a rise in body (core) temperature to 39.0–39.5°C during the first 2–4 h, and to sustained hyperthermia thereafter (38.5–39.0°C). After2 h of transient MCA occlusion hyperthermia could only be avoided if anesthesia (with control of temperature) was maintained for 2 h of ischemia and 1 h of recirculation or, in unanesthetized animals, if external cooling was maintained for 2 h of ischemia and 2 h of recirculation. Control of temperature only during ischemia did not prevent a postischemic rise in temperature. One hour of MCA occlusion had less effect on body temperature. Results are presented which suggest that the hyperthermia observed is due to an interference, by the intraluminal filament, of circulation to hypothalamic centers regulating body temperature. It is speculated that the hyperthermia induced may blunt or obliterate the effect of drugs, normally considered to ameliorate brain damage due to focal ischemia.     

Brown adipose tissue thermogenesis is activated by electrical and chemical (L-glutamate) stimulation of the ventromedial hypothalamic nucleus in cold-acclimated rats

Experiments were conducted to determine if both electrical and chemical stimulation of the ventromedial hypothalamic nucleus (VMH) could activate brown adipose tissue (BAT) thermogenesis. Age-matched, room-acclimated (21 degrees C) and cold-acclimated (4 degrees C for 3 weeks prior to testing) male Sprague-Dawley rats were given unilateral electrical or chemical stimulation to the VMH by way of a 'chemotrode apparatus'. The devised 'chemotrode' allowed both electrical stimulation (insulated piano wire stimulating electrode) and chemical stimulation (23 gauge stainless steel intracranial cannula of equal length) to be performed at the same VMH site using a common 19 gauge stainless steel outer guide tube. The first unilateral VMH electrical stimulation (0.5 ms pulse, 50 Hz and 120 microA for 30 s) caused no significant rise in interscapular brown adipose tissue temperature (TIBAT) colonic (Tc) or tail surface temperatures (Tt), compared to respective prestimulation control values in rats acclimated to 21 degrees C. In the 4 degrees C-acclimated group the first VMH electrical stimulation caused a significant rise in IBAT temperature. L-Glutamate administration to the same VMH site (60 nmol in 600 nl volume) also caused a significant increase in IBAT temperature in the 4 degrees C but not the 21 degrees C-acclimated rats. The rise in IBAT temperature following the L-glutamate injection to the 4 degrees C-acclimated group was similar to that found following the first electrical stimulation to this group. Interestingly, a second unilateral electrical stimulation of the VMH to 4 degrees C-acclimated rats could not evoke a similar increase in IBAT temperature suggesting that overall L-glutamate was acting in vivo as an excitotoxin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Targeting presynaptic norepinephrine transporter in brown adipose tissue: A novel imaging approach and potential treatment for diabetes and obesity

Brown adipose tissue (BAT) plays a significant role in metabolism. In this study, we report the use of atomoxetine (a clinically applicable norepinephrine reuptake inhibitor) for ¹⁸F‐FDG PET imaging of BAT and its effects on heat production and blood glucose concentration. Fasted‐male Sprague‐Dawley rats were administered with intravenous ¹⁸F‐FDG. The same rats were treated with atomoxetine (0.1 mg/kg, i.v.) 30 min before ¹⁸F‐FDG administration. To confirm the β‐adrenergic effects, propranolol (β‐adrenergic inhibitor) 5 mg/kg was given intraperitoneally 30 min prior to atomoxetine administration. The effect of atomoxetine on BAT metabolism was assessed in fasted and non‐fasted rats and on BAT temperature and blood glucose in fasted rats. In ¹⁸F‐FDG PET/CT images, interscapular BAT (IBAT) and other areas of BAT were clearly visualized. When rats were fasted, atomoxetine (0.1 mg/kg) increased the ¹⁸F‐FDG uptake of IBAT by factor of 24 within 30 min. Propranolol reduced the average ¹⁸F‐FDG uptake of IBAT significantly. Autoradiography of IBAT and white adipose tissue confirmed the data obtained by PET. When rats were not fasted, atomoxetine‐induced increase of ¹⁸F‐FDG uptake in IBAT was delayed and occurred in 120 min. For comparison, direct stimulation of β₃‐adrenreceptors in non‐fasted rats with CL‐316, 243 occurred within 30 min. Atomoxetine‐induced IBAT activation was associated with higher IBAT temperature and lower blood glucose. This was mediated by inhibition of norepinephrine reuptake transporters in IBAT leading to increased norepinephrine concentration in the synapse. Increased synaptic norepinephrine activates β₃‐adrenreceptors resulting in BAT hypermetabolism that is visible and quantifiable by ¹⁸F‐FDG PET/CT. Synapse, 2013. © 2012 Wiley Periodicals, Inc.     

Inhibition of interleukin-1β and prostaglandin E2 thermogenesis by glycyl-glutamine, a pro-opiomelanocortin-derived peptide

Interleukin-1β (IL-1β) and other cytokines produce fever by stimulating prostaglandin E₂ (PGE₂) synthesis in thermoregulatory regions of the preoptic area and anterior hypothalamus (POA/AH). Prostaglandin E₂ is thought to raise body temperature, at least in part, by stimulating β-endorphin release from pro-opiomelanocortin neurons that innervate the POA/AH. In this study, we investigated whether glycyl-glutamine (β-endorphin_30–31), an inhibitory dipeptide synthesized from β-endorphin post-translationally, inhibits IL-1β and PGE₂-induced hyperthermia. Hyperthermic sites were identified by microinjecting PGE₂ (3 fmol/1 μl) into the medial preoptic area (mPOA) of conscious, unrestrained rats. Interleukin-1β (1 U) injection into the same PGE₂ responsive thermogenic sites in the mPOA elicited a prolonged rise in colonic temperature (T_c) (+1.02±0.06°C) that persisted for at least 2 h. Glycyl-glutamine (3 nmol) co-injection into the mPOA inhibited IL-1β thermogenesis completely (T_c=−0.18±0.22°C). Glycyl-glutamine had no effect on body temperature when given alone to normothermic rats. Co-injection of individual amino acids, glycine and glutamine (3 nmol each amino acid), failed to influence IL-1β-induced thermogenesis, which indicates that Gly-Gln hydrolysis does not explain its inhibitory activity. Glycyl-glutamine (3 nmol) also prevented the rise in body temperature produced by PGE₂ (PGE₂=0.89±0.05°C; PGE₂ plus Gly-Gln=−0.16±0.14°C), consistent with evidence that PGE₂ mediates IL-1β-induced fever. These findings demonstrate that Gly-Gln inhibits the thermogenic response to endogenous pyrogens.     

Hyperthermic reactions induced by orexin A: role of the ventromedial hypothalamus

This experiment tested the involvement of the ventromedial hypothalamus (VMH) in the sympathetic and hyperthermic reactions induced by an intracerebroventricular (i.c.v.) injection of orexin A (1.5 nmol). In the first part of the experiment, the firing rate and cytochrome oxidase activity of the VMH neurons, and the colonic temperature were monitored in 12 urethane-anaesthetized rats before an i.c.v. injection of orexin and over a period of 2 h after the injection. Orexin induced an increase in the firing rate, colonic temperature and cytochrome oxidase activity. A group of 12 rats was used as a control: saline, but not orexin, was injected. No modifications in the firing rate, cytochrome oxidase reactivity and colonic temperature were noted. In the second part of the experiment, 12 rats were anaesthetized and lesioned bilaterally in the VMH with an injection of ibotenic acid. Sham lesions were carried out in 12 control rats. After 48 h, all animals were anaesthetized with ethyl-urethane. The firing rates of the sympathetic nerves to interscapular brown adipose tissue (IBAT), along with IBAT and colonic temperatures and heart rate were monitored before and over a period of 2 h after an i.c.v. injection of orexin or saline in the lesioned and sham-lesioned rats. Orexin increased the sympathetic firing rate, IBAT and colonic temperatures and heart rate in the sham-lesioned rats. These increases were reduced by lesion of VMH. Saline did not induce any modification. These findings indicate that the VMH is involved in the control of the orexin-induced hyperthermia.     

Effects of diazepam on body temperature of the aged squirrel monkey

Hypothermia was produced by IM administration of diazepam (0.125–0.5 mg/kg) to squirrel monkeys of various ages (2–16 years) in a thermoneutral (23 ± 0.5°C) environment with animals over eight years of age having slightly greater responses. Hypothermia caused by an intermediate dose (0.25 mg/kg) was augmented in a cold environment (15°C), especially in the older animals. There was no marked alteration in the temperature change/age regression after 0.25 mg/kg diazepam in a hot environment (30°C) compared with the control response. Injections of diazepam (1.25-5.0 /μg) into the lateral cerebral ventricle in a thermoneutral environment produced hyperthermia rather than hypothermia in all animals, and the magnitude of the induced hyperthermia was smaller in older monkeys. The results support previous case reports in man and suggest that this commonly used drug can induce hypothermia, especially in older primates exposed to cold. The drug action responsible for this temperature change appears to take place peripherally rather than within the brain.     

Insulin co-injection suppresses the thermogenic response to glutamate microinjection into the VMH in rats

Selective stimulation of ventromedial hypothalamic nucleus (VMH) neurons by microinjection of the excitatory amino acid glutamate sharply increased interscapular brown adipose tissue (IBAT) and core temperatures in urethane-anaesthetized rats. This effect was blocked by co-injection of insulin (0.1-1 microgram) though not an inactive insulin analog, TNB3 insulin. Injection of insulin (1 microgram) into the contralateral VMH or systemic administration of insulin (1 microgram) had no effect on the thermogenic response to intra-VMH glutamate. These results complement those showing that intra-VMH insulin suppresses the basal firing rate of sympathetic nerves to IBAT and diminishes cold-induced nonshivering thermogenesis in BAT and add support to the view that insulin functions as an inhibitory signal on VMH neurons controlling thermogenesis in BAT.     

ACh and 5-HT stimulated thermogenesis at different core temperatures in the He-Cold hypothermie hamster

Hamsters in deep experimentally induced hypothermia, at body temperatures between 7°C and 11.5°C, were microinjected with 5-HT and ACh at brain sites in the anterior-preoptic area of the hypothalamus (AH/POA). ACh or 5-HT was injected into an AH/POA site at different starting core temperatures in different groups of hypothermic hamsters. Colonic temperatures (Tc) were maintained, following He-Cold induction, in a temperature controlled environmental chamber and measured with a YSI thermister probe and YSI tele-thermometer. Injections of either 5-HT or ACh at Tc's between 7.0°C and 9.0°C elicited only modest increases in Tc i.e., 0.3°C–0.6°C, respectively. As Tc increased, however, to ranges between 9.1°C–10.0°C and in different animals to greater than 10°C both ACh and 5-HT at the same sites elicited significant increases in Tc, 1.5°C for 5-HT and 2.2°C for ACh compared to saline injections. These data suggest that at the lowest Tc's we are observing a “cold block” of temperature sensitive sites in the AH/POA. Increasing the starting Tc beyond 9.0°C however, evokes significant increases in heat-gain following AH/POA injection of either ACh or 5-HT. These data are consistent with Myers' observations concerning the organization of heat-gain mechanisms at AH/POA sites. In addition, they suggest that both the afferent limb of the heat-gain circuit (5-HT) and the efferent limb of the circuit (ACh) are functionally impaired when Tc is close to the physiological limit in the He-Cold hypothermic hamster.     

Disinhibition of lower midbrain neurons enhances non-shivering thermogenesis in anesthetized rats

The hypothesis that the lower midbrain, specifically in and around the retrorubral field (RRF) and/or rubrospinal tract (rs), contains a tonic inhibitory mechanism on non-shivering thermogenesis (NST) was examined in urethane-anesthetized rats. It has been proposed that removal of the tonic inhibitory mechanism in the lower midbrain causes body temperature increase through disinhibition-induced activation of the central sympathetic nervous system. The present experiments were carried out to examine whether and where the proposed midbrain region contains cell bodies that tonically inhibit the NST, and if so, whether they receive any influence from the hypothalamus. Male Wistar rats were anesthetized with urethane (1. 0-1.2 g/kg, i.p.), and various agents were microinjected into the RRF and rs areas of one side before and after knife-cut in the other side of the lower midbrain or isolation of the hypothalamus from the midbrain. Changes in interscapular brown adipose tissue (IBAT), rectum, and tail skin temperatures were monitored. RESULTS: (1) unilateral midbrain procaine increased IBAT and rectal temperatures only after un-injected side of the midbrain had been pre-transected. (2) Effective midbrain sites for procaine to increase IBAT and rectal temperatures was laterally extended. (3) Tetrodotoxin microinjected into the midbrain site where procaine increased IBAT and rectal temperatures also raised both temperatures. (4) l-glutamate decreased IBAT and rectal temperatures when microinjected into one of the most inner midbrain area of procaine-sensitive sites without affecting tail skin temperature. (5) Isolation of the hypothalamus from the lower midbrain did not affect midbrain procaine-induced IBAT and rectal temperature increases. These results suggest that neurons that tonically inhibit the NST are located in the area close to the midline adjacent to the RRF and rs, and that the integrity of the neurons to tonically inhibit the NST is not affected by disconnecting the hypothalamus from the midbrain.     

Temperature regulation and dopaminergic systems in the brain: Does the substantia nigra play a role?

Male Sprague-Dawley rats (250–300 g) were stereotaxically implanted above the substantia nigra (SN) and preoptic/anterior hypothalamus (PO/AH) with 23 gauge stainless-steel guide tubes. Microinjections of apomorphine (APO), pimozide, or 0.9% saline were made bilaterally in 0.5 or 1.0 μl vols. using a Harvard infusion pump. Oxygen consumption, and colonic (T_c), tail-skin, and ambient temperatures were monitored each minute. Microinjections of APO into the SN produced a dose-dependent hypothermia that was antagonized by the central (1.0 μg) or systemic (0.5 mg/kg i.p.) injection of pimozide. This hypothermia was associated with increased heat loss and decreased heat production and occurred at ambient temperatures of 15, 23 and 35°C indicating that APO did not produce a poikilothermic state. Injecting 20 μg APO into the PO/AH or SN produced similar hypothermic responses; T_c fell 0.87± 0.10°C and1.02 ± 0.08°C, respectively. Pimozide injected into the SN failed to alter thermoregulation during exercise or exogenous heating. Moreover, systemic injections of APO before and after electrolytic lesioning of the SN produced similar hypothermic responses. However, when the SN was lesioned, (a) resting T_c was significantly reduced, and (b) during exposure to a 35°C environment for 55 min, T_c lose to39.5 ± 0.68°C before the lesion compared with a rise to only38.5 ± 0.13°C after the lesion. We conclude that the pharmacological data implicate a thermoregulatory role for dopamine receptors in the SN of the rat, but the functional significance of this central location in temperature regulation remains to be elucidated.