Fasting-induced Torpor Research Articles

CLIC5 mutant mice are resistant to diet-induced obesity and exhibit gastric hemorrhaging and increased susceptibility to torpor

Chloride intracellular channel 5 (CLIC5) and other CLIC isoforms have been implicated in a number of biological processes, but their specific functions are poorly understood. The association of CLIC5 with ezrin and the actin cytoskeleton led us to test its possible involvement in gastric acid secretion. Clic5 mutant mice exhibited only a minor reduction in acid secretion, Clic5 mRNA was expressed at only low levels in stomach, and Clic5 mutant parietal cells were ultrastructurally normal, negating the hypothesis that CLIC5 plays a major role in acid secretion. However, the mutants exhibited gastric hemorrhaging in response to fasting, reduced monocytes and granulocytes suggestive of immune dysfunction, behavioral and social disorders suggestive of neurological dysfunction, and evidence of a previously unidentified metabolic defect. Wild-type and mutant mice were maintained on normal and high-fat diets; plasma levels of various hormones, glucose, and lipids were determined; and body composition was studied by quantitative magnetic resonance imaging. Clic5 mutants were lean, hyperphagic, and highly resistant to diet-induced obesity. Plasma insulin and glucose levels were reduced, and leptin levels were very low; however, plasma triglycerides, cholesterol, phospholipids, and fatty acids were normal. Indirect calorimetry revealed increased peripheral metabolism and greater reliance on carbohydrate metabolism. Because Clic5 mutants were unable to maintain energy reserves, they also exhibited increased susceptibility to fasting-induced torpor, as indicated by telemetric measurements showing episodes of reduced body temperature and heart rate. These data reveal a requirement for CLIC5 in the maintenance of normal systemic energy metabolism.

Cardiovascular changes during daily torpor in the laboratory mouse

The laboratory mouse is a facultative daily heterotherm in that it experiences bouts of torpor under caloric restriction. Mice are the most frequently studied laboratory mammal, and often, genetically modified mice are used to investigate many physiological functions related to weight loss and caloric intake. As such, research documenting the cardiovascular changes during fasting-induced torpor in mice is warranted. In the current study, C57BL/6 mice were implanted either with EKG/temperature telemeters or blood pressure telemeters. Upon fasting and exposure to an ambient temperature (T(a)) of 19 degrees C, mice entered torpor bouts as assessed by core body temperature (T(b)). Core T(b) fell from 36.6 +/- 0.2 degrees C to a minimum of 25.9 +/- 0.9 degrees C during the fast, with a concomitant fall in heart rate from 607 +/- 12 beats per minute (bpm) to a minimum of 158 +/- 20 bpm. Below a core T(b) of 31 degrees C, heart rate fell exponentially with T(b), and the Q(10) was 2.61 +/- 0.18. Further, mice implanted with blood pressure telemeters exhibited similar heart rate and activity profiles as those implanted with EKG/temperature telemeters, and the fall in heart rate and core T(b) during entrance into torpor was paralleled by a fall in blood pressure. The minimum systolic, mean, and diastolic blood pressures of torpid mice were 62.3 +/- 10.2, 51.9 +/- 9.2, 41.0 +/- 7.5 mmHg, respectively. Torpid mice had a significantly lower heart rate (25-35%) than when euthermic at mean arterial pressures from 75 to 100 mmHg, suggesting that total peripheral resistance is elevated during torpor. These data provide new and significant insight into the cardiovascular adjustments that occur in torpid mice.

The pharmacology and molecular mechanisms underlying temperature regulation and torpor

The ability to enter a hypometabolic state upon restriction of caloric intake is pivotal for animal survival: balancing the energy budget in endotherms can be a real struggle when food is not available and/or the demand for heat production to maintain homeothermy becomes excessive. Bouts of torpor, characterized by metabolic rates well below a basal metabolic rate and core body temperatures that may be just a few degrees above the ambient temperature, are utilized among many organisms across the animal kingdom, including those that could be described as typical laboratory animals, like the mouse or hamster. Daily heterotherms, which are the focus of this commentary, enter shallow torpor bouts and do so usually under acute food shortage conditions and a relatively cool environment. Due to their small size, the body temperature of these animals is very responsive to food deprivation, pharmacological inhibition of metabolic rate, and cardiovascular depressants. This commentary examines recent developments concerning the neuroendocrine mechanisms in place that may mediate fasting-induced torpor in daily heterotherms. Further this commentary highlights pharmacological induction of hypothermia in small mammals.

AMP does not induce torpor

Torpor, a state characterized by a well-orchestrated reduction of metabolic rate and body temperature (T(b)), is employed for energetic savings by organisms throughout the animal kingdom. The nucleotide AMP has recently been purported to be a primary regulator of torpor in mice, as circulating AMP is elevated in the fasted state, and administration of AMP causes severe hypothermia. However, we have found that the characteristics and parameters of the hypothermia induced by AMP were dissimilar to those of fasting-induced torpor bouts in mice. Although administration of AMP induced hypothermia (minimum T(b) = 25.2 +/- 0.6 degrees C) similar to the depth of fasting-induced torpor (24.9 +/- 1.5 degrees C), ADP and ATP were equally effective in lowering T(b) (minimum T(b): 24.8 +/- 0.9 degrees C and 24.0 +/- 0.5 degrees C, respectively). The maximum rate of T(b) fall into hypothermia was significantly faster with injection of adenine nucleotides (AMP: -0.24 +/- 0.03; ADP: -0.24 +/- 0.02; ATP: -0.25 +/- 0.03 degrees C/min) than during fasting-induced torpor (-0.13 +/- 0.02 degrees C/min). Heart rate decreased from 755 +/- 15 to 268 +/- 17 beats per minute (bpm) within 1 min of AMP administration, unlike that observed during torpor (from 646 +/- 21 to 294 +/- 19 bpm over 35 min). Finally, the hypothermic effect of AMP was blunted with preadministration of an adenosine receptor blocker, suggesting that AMP action on T(b) is mediated via the adenosine receptor. These data suggest that injection of adenine nucleotides into mice induces a reversible hypothermic state that is unrelated to fasting-induced torpor.

Fasting‐induced torpor requires β3 adrenergic receptor signaling

Torpor, a controlled rapid drop in metabolic rate and body temperature (Tb), is a hypometabolic adaptation to stressful environmental conditions, which occurs in many small mammals, marsupials, and birds. To date, signaling pathways required for torpor have not been identified. We examined the role of the sympathetic nervous system (SNS) in mediating the torpor adaptation to fasting by telemetrically monitoring the Tb of dopamine β-hydroxylase knockout (Dbh) −/− mice which lack the ability to produce the SNS transmitters, norepinephrine (NE) and epinephrine (Epi). Control (Dbh +/−) mice readily reduced serum leptin levels and entered torpor after a fast in a cool environment. In contrast, Dbh −/− mice failed to reduce serum leptin and enter torpor under fasting conditions, whereas restoration of peripheral but not central NE lowered serum leptin levels and rescued the torpor response. Torpor was expressed in fasted Dbh −/− mice immediately after administration of either the non-selective β adrenergic receptor agonist, isoproterenol, or the β3 AR specific agonist, CL316243, but not after administration of β1, β2, or α1 agonists. Importantly, a β3-specific antagonist severely blunted fasting-induced torpor in control mice, whereas other AR antagonists were ineffective. These results define a critical role of peripheral SNS activity at β3 AR-containing tissues in the torpor adaptation.

The Full Expression of Fasting-Induced Torpor Requires β3-Adrenergic Receptor Signaling

Torpor, a controlled rapid drop in metabolic rate and body temperature (Tb), is a hypometabolic adaptation to stressful environmental conditions, which occurs in many small mammals, marsupials, and birds. To date, signaling pathways required for torpor have not been identified. We examined the role of the sympathetic nervous system (SNS) in mediating the torpor adaptation to fasting by telemetrically monitoring the Tb of dopamine beta-hydroxylase knock-out (Dbh-/-) mice, which lack the ability to produce the SNS transmitters, norepinephrine (NE), and epinephrine. Control (Dbh+/-) mice readily reduced serum leptin levels and entered torpor after a fast in a cool environment. In contrast, Dbh-/- mice failed to reduce serum leptin and enter torpor under fasting conditions, whereas restoration of peripheral but not central NE lowered serum leptin levels and rescued the torpor response. Torpor was expressed in fasted Dbh-/- mice immediately after administration of either the nonselective beta-adrenergic receptor agonist isoproterenol or the beta3-adrenergic receptor (AR)-specific agonist CL 316243 [disodium (RR)-5-[2-[[2-(3-chlorophenyl)-2-hydroxyethyl]-amino]propyl]-1,3-benzodioxazole-2,2-dicarboxylate], but not after administration of beta1, beta2, or alpha1 agonists. Importantly, the beta3-specific antagonist SR 59230A [3-(2-ethylphenoxy)-1-[(1,S)-1,2,3,4-tetrahydronapth-1-ylamino]-2S-2-propanol oxalate] severely blunted fasting-induced torpor in control mice, whereas other AR antagonists were ineffective. These results define a critical role of peripheral SNS activity at beta3-AR-containing tissues in the torpor adaptation to limited energy availability and cool ambient temperature.

Torpor in mice is induced by both leptin-dependent and -independent mechanisms.

We tested the effect of chronic leptin treatment on fasting-induced torpor in leptin-deficient A-ZIP/F-1 and ob/ob mice. A-ZIP/F-1 mice have virtually no white adipose tissue and low leptin levels, whereas ob/ob mice have an abundance of fat but no leptin. These two models allowed us to examine the roles of adipose tissue and leptin in the regulation of entry into torpor. Torpor is a short-term hibernation-like state that allows conservation of metabolic fuels. We first characterized the A-ZIP/F-1 animals, which have a 10-fold reduction in total body triglyceride stores. Upon fasting, A-ZIP/F-1 mice develop a lower metabolic rate and decreased plasma glucose, insulin, and triglyceride levels, with no increase in free fatty acids or beta-hydroxybutyrate. Unlike control mice, by 24 hr of fasting, they have nearly exhausted their triglycerides and are catabolizing protein. To conserve energy supplies during fasting, A-ZIP/F-1 (but not control) mice entered deep torpor, with a minimum core body temperature of 24 degrees C, 2 degrees C above ambient. In ob/ob mice, fasting-induced torpor was completely reversed by leptin treatment. In contrast, neither leptin nor thyroid hormone prevented torpor in A-ZIP/F-1 mice. These data suggest that there are at least two signals for entry into torpor in mice, a low leptin level and another signal that is independent of leptin and thyroid hormone levels. Studying rodent torpor provides insight into human torpor-like states such as near drowning in cold water and induced hypothermia for surgery.

Fasting-induced torpor in Mus musculus and its implications in the use of murine models for human obesity studies

1. 1. Fasting at room temperature for 24–48 hr can induce shallow daily torpor in mice of several strains. 2. 2. Genetically-obese (ob/ob) C57B1/6 mice show torpor even in the presence of food. 3. 3. Chemically-obese (gold-thioglucose or bipiperidyl mustard) mice of C57B1/6 and T/O strains appear resistant to torpor. 4. 4. During cold acclimation (3–5°) fasting also induces torpor in lean mice, with gradually improving rates of arousal. 5. 5. In acute cold shock (3–5°), previously weight-reduced ob/ob mice show resistance to change in body temperature, while control ob/ob mice drift immediately towards hypothermia.

Seasonal spontaneous torpor in the white-footed mouse, Peromyscus leucopus

1. 1. In winter spontaneous torpor occured only in mice that had been acclimated to cold weather the preceding fall. 2. 2. Daily spontaneous torpor was very frequent from late fall to early spring. Minimum thoracic temperatures averaged 17·3°C and minimum heart rate was 60 beats/min. 3. 3. All mice became torpid when fasted in late spring and summer. Fasting-induced torpor differed from spontaneous torpor in a slower rate of arousal, higher minimum thoracic temperature and irregular heart rhythm.