Min Of Cold Exposure Research Articles

209-OR: Evidence that Agrp Neuron Activation Drives Cold-Induced Hyperphagia

Although hyperphagic feeding during cold exposure is widely viewed as a response to the negative energy state that results from increased thermogenesis, the underlying mechanisms are unknown. We investigated an alternative hypothesis, that during cold exposure, hyperphagia is mounted in anticipation of homeostatic need, rather than as a compensatory response. In support of this hypothesis, we report that in chow-fed mice acutely housed at 14°C, increases of food intake and thermogenesis occur both rapidly and simultaneously, implying hyperphagia is not mounted as a compensatory response. We next asked whether this cold-induced feeding response involves activation of agouti-related peptide (Agrp) neurons, which are implicated in other forms of hyperphagic feeding. Consistent with this hypothesis, we report that Agrp neurons are activated within 1-2 min of cold exposure and prior to the onset of hyperphagia, as judged by in vivo fiber photometry and confirmed by cFos induction (Agrp+/cFos+, 22°C: 5.3±4.5% vs. 14°C: 20.2±6.1%; p<0.01). To test whether Agrp neuron activation is required for cold-induced hyperphagia, we employed a chemogenetic strategy to silence Agrp neurons. As predicted, their silencing prevented hyperphagic feeding (2h food intake after housing at 14°C: 0.26±0.06 g vs. 0.10±0.04 g; p<0.05), but not the associated thermogenic response (0.63 vs. 0.62 kcal/hr; p=ns) during cold exposure. Interestingly, cold-induced hyperphagia is absent in mice with diet-induced obesity (DIO). Consequently, unlike chow-fed mice, these animals lose weight during cold exposure. These observations suggest that 1) Agrp neurons lie downstream of thermoregulatory neurocircuits, 2) during cold exposure, increased Agrp neuronal firing occurs rapidly as part of a homeostatic response that preserves body temperature and energy stores, and 3) this response appears to be impaired in mice with DIO. Future studies will map the neurocircuitry underlying these responses and clarify how DIO impairs their activation. Disclosure J.D. Deem: None. C.L. Faber: None. C. Pedersen: None. B.N. Phan: None. K. Ogimoto: None. S.A. Larsen: None. M.A. Tran: None. V. Damian: None. K. Kaiyala: None. J. Scarlett: None. M.R. Bruchas: None. M.W. Schwartz: Research Support; Self; Novo Nordisk A/S. G.J. Morton: Research Support; Self; Novo Nordisk A/S. Funding American Diabetes Association (1-19-PDF-103 to J.D.D.); Dick and Julia McAbee Foundation; National Institutes of Health (T32-HL007028-39, R01DK089056, R01DK101997, R01DK124238, R01DK089056, P30DK017047, P30DK035816)

Thermal Sensations during a Partial-Body Cryostimulation Exposure in Elite Basketball Players.

Partial-body cryostimulation is used to improve recovery after exercise, especially during competitions or heavy training; however, a limited number of studies have been conducted with international-level athletes in situ during competitions. This study was undertaken to assess the thermal sensation ratings during 3 min of cold exposure (at –130°C) in 24 international-level athletes during the European Basketball Championship. The mean thermal sensation score, measured using a perceptive scale, increased significantly (p < 0.05) during partial-body cryostimulation exposure in athletes from 3.0 ± 1.7 at 30 s to 5.7 ± 2.3 at 3 min (maximal observed value = 10.0). The mean value of 5.7 is considered a “cold” sensation on the scale (ranging from 0 = neutral sensation to 10 = very cold). However, we observed a large inter-individual variation in the perceived thermal sensations. The body mass index was significantly and negatively correlated with the thermal sensation value after 2 min 30 s and 3 min of exposure in females (r = –0.61, n = 13, p < 0.05; r = –0.56, n = 13, p = 0.054, respectively). Three participants reported high perceived thermal sensation after 30 s of exposure and their cold-induced discomfort worsened as the exposure continued. In conclusion, a 3-min exposure is globally well tolerated by athletes and can be used during a heavy competition period and/or during a training period. However, special attention should be given to female athletes with a low body mass index as they seem to be much more sensitive to cold.

Infrared thermal imaging as a method to study thermogenesis in the neonatal lamb

The combination of heat generation and reducing heat loss from the skin surface is important for maintaining core body temperature in a neonate. Thermogenesis studies traditionally focus on measurement of core body temperature but not the contribution of radiated heat loss at the skin surface. This study aimed to evaluate the utility of using thermal imaging to measure radiated heat loss in newborn lambs. Continuous thermal images of newborn lambs were captured for 30 min each during the baseline (11−18°C), cold-exposure (0°C) and recovery (11−18°C) periods by using an infrared camera. Core body temperature measured by rectal thermometer was also recorded at the end of each period. In all, 7 of the 10 lambs evaluated had reduced rectal temperatures (0.4−1°C) between the baseline and recovery periods, while three maintained body temperature despite cold exposure. During the baseline period, infrared heat loss was relatively stable, followed by a rapid decrease of 5°C within 5 min of cold exposure. Heat loss continued to decrease linearly in the cold-exposure period by a further 10°C, but increased rapidly to baseline levels during the recovery period. A temperature change of between 20°C and 35°C was observed during the study, which was likely to be due to changes in vasoconstriction in the skin to conserve heat. The present study has highlighted the sensitivity of infrared thermal imaging to estimate heat loss from the skin in the newborn lamb and shown that rapid changes in heat loss occur in response to cold exposure.

Regulation and characterization of four CBF transcription factors from Brassica napus.

Four orthologues of the Arabidopsis CBF/Dreb transcriptional activator genes were isolated from the winter Brassica napus, cv. Jet neuf. All four BNCBF clones encode a putative DRE/CRT (LTRE)-binding protein with an AP2 DNA-binding domain, a putative nuclear localization signal and a possible acidic activation domain. Deduced amino acid sequences suggested that BNCBFs 5, 7and 16 are very similar to the Arabidopsis CBFI whereas BNCBF17 is different in that it contains two extra regions of 16 and 21 amino acids in the acidic domain. Transcripts hybridizing specifically to BNCBF17 and to one or more of the other BNCBFs accumulated in leaves within 30 min of cold exposure of the Brassica seedlings and preceded transcript accumulation of the cold-inducible BN28 gene, a Brassica orthologue of the cor6.6 or KIN gene from Arabidopsis. Cold-induced accumulation of BNCBF17 mRNA was rapid but was short-lived compared to transcripts hybridizing to BNCBF5/7/16. Transcripts hybridizing to one or more of BNCBF5/7/16 accumulated at low levels after the plants were subjected to prolonged exposure to salt stress. BNCBF17 was not responsive to salt stress. BNCBF transcript accumulation was similar in both spring and winter Brassica but the persistence of the transcripts in the cold were generally shorter in the spring than in the winter type. BNCBF5 and 17 proteins bind in vitro to the LTRE domains of the cold-inducible BN115 (cor15a orthologue) or BN28 promoters. Differential binding preferences, however, to LTREs between BNI 15 and BN28 were observed. Mutation of the core CCGAC sequence of the LTRE indicated that BNCBF17 had a lower sequence binding specificity than BNCBF5. Furthermore, experiments indicated that the LTREs were able to drive BNCBF5 and 17 trans-activation of the Lac-Z reporter gene in yeast. We conclude that the BNCBFs reported here could function as trans-acting factors in low-temperature responses in Brassica, controlling the expression of cold-induced genes through an ABA-independent pathway.

Training reduces autonomic cardiovascular responses to both exercise-dependent and -independent stimuli in humans

Training attenuates the sympathetic pressor response to dynamic exercise. However, it is uncertain how training alters other patterns of cardiovascular autonomic activation. Therefore, we have quantified circulatory responses to a series of standard autonomic tests in highly fit and unfit subjects and examined the effects of a short-term training programme on these responses. Subjects were defined as either unfit ( n=8) or fit ( n=8) on the basis of training history and a maximal fitness test (VO 2peak 54±2.3 cf. 68±2.8 (ml min −1) kg −1, means±S.E.M., P<0.05). On a separate day, the blood pressure, heart rate and forearm vascular conductance responses to a sustained handgrip to fatigue, 2 min mental arithmetic and 2 min of cold exposure were measured. All stimuli were associated with elevated blood pressures and heart rates, but these responses were significantly attenuated in the trained group. In the untrained subjects, forearm vascular conductance increased during exercise (from 0.032±0.004 to 0.05±0.007 (ml min −1) 100 ml −1 mm Hg −1, P<0.05) and during mental arithmetic (from 0.028±0.003 to 0.04±0.006 (ml min −1) 100 ml −1 mm Hg −1, P<0.05), but trained subjects showed no rise in conductance during either test. All untrained subjects undertook a moderate intensity 5-week training programme, which significantly increased VO 2peak (54±2.3 to 57±2 (ml min −1) kg −1, P<0.05). Qualitatively similar blunting of pressor, tachycardic and vasodilator responses were seen in this group post-training. These results demonstrate that the blunting of sympathetic vasomotor activation that follows training is not restricted to reflexes associated with exercise, and does not depend on training being prolonged or intense.

Activation of bulbospinal serotonergic neurons during cold exposure

In a four-part study, we expand on our previous report that bulbospinal serotonin (5HT) neuronal activation occurs with 24 h of cold exposure. To characterize temporal aspects, rats were exposed to 3 degrees C or were maintained at 22 degrees C for 2, 8, 48, or 96 h (experiment 1) or for 15, 30, or 60 min (experiment 2). To ensure that cold-induced changes in 5HT activity were not due to disturbances in diurnal pattern, rats in experiment 3 were exposed to cold (8 h) during the dark cycle. To explore the hypothesis that cold-induced 5HT activation is part of a broad metabolic response that includes activation of the sympathetic nervous system, metabolically impaired (hypothyroid) rats were exposed to 8 degrees C in experiment 4. Significant increments in 5-hydroxyindoleacetic acid (SHIAA) concentration were evident by 60 min of cold exposure and existed at all later time points measured. These findings were most robust in spinal cord and rostral brainstem. Activation in spinal cord was also found when rats were exposed to 8 h of cold during the dark cycle, the active period for rats. In experiment 4, hypothyroid rats exhibited significantly greater norepinephrine excretion compared with control rats exposed to the same cold stimulus; this finding was accompanied by significantly greater increments in 5HIAA concentration in rostral brainstem and spinal cord of hypothyroid rats. In addition, significant elevations in tryptophan concentration were noted throughout the brainstem and spinal cord of cold-exposed, hypothyroid rats relative to room temperature, hypothyroid rats. This finding suggested that elevations in 5HIAA concentration in these rats were due to increases in precursor availability. The implications of these findings relative to autonomic and metabolic control are discussed.

Interaction between cold and hypoxia on pulmonary circulation in COPD.

This study was designed to investigate the interaction of mild and localized cold exposure and hypoxia on pulmonary hemodynamics in chronic obstructive pulmonary disease (COPD). Nineteen patients with COPD were studied at sea level and seven at an altitude of 2,640 m. For all patients, pulmonary hemodynamic measurements were performed 10 min after insertion of a catheter in a femoral vein and following 10 min of cold exposure. Cold exposure was restricted to the forehead, and subjects breathed air at ambient temperature. Flow and temperature of air (1.5 L.s(-1), 5 degrees C) to the forehead were chosen to cool down the forehead skin to approximately 20 degrees C without discomfort for the subject. For the seven patients studied at high altitude, the same measurements were also performed after 5 min of oxygen supplementation with and without cold exposure. At sea level, an increase in pulmonary vascular resistance (PVR) during cold exposure was inversely related to the initial PaO2. In six severe hypoxic subjects (PaO2 < 50 mm Hg), PVR increased by 24%. At high altitude, PVR was significantly increased by 15%. After O2 supplementation, cold exposure did not induce an increase in PVR. We concluded that mild and localized cold exposure to the forehead only induced an increase in PVR in COPD patients with severe hypoxia. Moreover, in cold exposure responders, O2 supplementation negated the effect of cold exposure on pulmonary hemodynamics.

Rapid Changes in Somatostatin and TRH mRNA in Whole Rat Hypothalamus in Response to Acute Cold Exposure

Acute cold stimulus induces activation of the thyreotropic axis characterized by a rapid increase in plasma thyrotropin (TSH). Since pituitary TSH release is mainly regulated by two hypothalamic hormones: thyrotropin-releasing hormone (TRH) and somatostatin, the aim of this study was to analyse whether changes in the steady state mRNA levels and peptide content of these neurohormones occur under acute cold stimulation in rats. Northern blot analysis of hypothalamic somatostatin mRNA levels after 15, 30, 60 or 180 min of cold exposure revealed a 2.0-fold increase after 15 min at 4 degrees C. This augmentation was followed by a return to control values at 30 min. However, the hypothalamic content of somatostatin was not significantly modified at any cold exposure time. TRH mRNA showed a similar pattern to somatostatin, with a 2.5-fold increase after 15 min at 4 degrees C. In contrast, hypothalamic TRH content was significantly decreased after 15 min cold exposure, returning to control values at 30 min. The increase in mRNA levels was specific for the two hypothalamic hormones, since there was no concomitant variation in GAPDH mRNA used as negative control. These results suggest that the organism is quickly aroused by cold stimulus, triggering rapid activation in transcription of the two neurohormones involved in the regulation of the thyreotrope axis. Since the peptide contents did not show the same pattern, a quantitative change in transcription or in mRNA stability does not appear to be a prerequisite for increased peptide expression, suggesting that somatostatin and TRH gene expressions could be regulated at translational or post-translational steps.

Hind leg muscle amino acid balances in cold-exposed rats.

The changes in hind leg tissue (muscle and skin) amino acid pool size and arteriovenous balance were measured in rats subjected to 0-90 min of cold exposure (4 degrees C). Tissue free amino acid pools presented a different composition pattern from protein amino acids. Muscle rapidly reacted to cold exposure by releasing small amounts of some amino acids (alanine, aspartate), with only small changes in pool size during the first 30 min. Amino acid oxidation was very limited during the whole period of cold exposure, since at all times tested there was either nil ammonia efflux or net absorption of ammonia and glutamine; i.e. the muscle was in positive nitrogen balance throughout the period studied. Thus most of the amino acid nitrogen taken up from the blood and not found in the free amino acid pools must have been incorporated into protein, since it was not oxidized, as shown by the glutamine and ammonia balance. The data on amino acid incorporation into proteins indicate that hind leg protein turnover is rapidly and widely modulated from a low initial setting upon cold exposure to a higher protein synthesis rate immediately afterwards, suggesting that protein turnover may have an important role in short-term events in cold-exposed muscle, in addition to its influence in long-term adaptation.

Seasonal variations in the responses of glycolytic intermediates of human erythrocytes to acute cold exposure.

Seven male students were studied to observe the effects of acute cold exposure (at 10°C for 60 min) on erythrocyte concentrations of glycolytic intermediates in summer and in winter. The subjects shivered slightly but frankly in both experiments. Significant decreases were observed in the concentrations of pyruvate and lactate during body cooling in summer, but not in winter. The lactate concentration remained significantly reduced 15 min after cold exposure. After 60 min of cold exposure in summer, a negative crossover point appeared to exist between phosphoenolpyruvate and pyruvate and erythrocyte pyruvate kinase activity showed a significant decrease. No seasonal difference was observed in the initial control values of the intermediates measured. From these results and the fact that glucose, pyruvate and lactate are evenly distributed between erythrocytes and plasma, it is likely that erythrocytes and skeletal muscles need less fuel substrate, glucose during cold exposure in winter than in summer, suggesting that an increased economy of energy for homeostasis is achieved.