Diet-induced obesity and cardiovascular regulation in C57BL/6J mice.

1. In the present study, we determined the effect of diet-induced obesity on cardiovascular and metabolic regulation in mice at standard laboratory temperatures (ambient temperature (Ta) = 22 degrees C) and during exposure to thermoneutrality (Ta = 30 degrees C). 2. Male C57BL/6J (B6) mice fed a high-fat diet (HFF; n = 17) or chow (CHW; n = 14) for 15 weeks were surgically instrumented with telemetry devices, housed in metabolic chambers and assigned to either control or atenolol treatment (25 mg/kg per day in drinking water) to determine the effects of obesity on baseline cardiovascular function and on the responses to thermoneutrality and 24 h fasting. Mean arterial pressure (MAP), heart rate (HR), arterial pressure and HR variability (time and frequency domain), oxygen consumption (VO2) and locomotor activity were determined. 3. The HFF mice exhibited increased bodyweight (+10.6 +/- 4.1 g), moderate light period hypertension (+8.6 +/- 2.6 mmHg), no difference in HR and increased HR variability at standard laboratory temperature compared with CHW controls. Atenolol produced less of a decrease in HR in HFF mice (-42 +/- 10 b.p.m.) compared with CHW controls (-73 +/- 15 b.p.m.). Acute exposure to thermoneutrality (Ta = 30 degrees C) reduced HR similarly in both HFF and CHW mice (approximately 175 b.p.m.), but reduced MAP less in HFF than in CHW mice (-7.3 +/- 2.5 and -15.2 +/- 1.0 mmHg), respectively. Atenolol treatment had only minor effects on the HR response to thermonuetrality (-114 +/- 13 and -129 +/- 8 b.p.m. in HFF and CHW mice, respectively). The HFF mice displayed greater fasting-induced reductions in light period MAP than did CHW mice (-10.0 +/- 1.1 vs-3.1 +/- 3.5 mmHg, respectively), whereas HR was decreased equally in both groups. Fasting-induced increases in HR variability were attenuated in HFF mice. 4. We conclude that diet-induced obesity produced generally minor changes in cardiovascular regulation in B6 mice at baseline, some of which are distinct from the effects of diet-induced obesity in larger animal models. In contrast, acute variations in Ta or caloric availability produce pronounced alterations in cardiovascular function in either lean or obese mice, which are generally evident after atenolol and, thus, presumably not due exclusively to variation in cardiac sympathetic activity. Interestingly, the degree of obesity induced hypertension was augmented when mice were studied at thermonuetrality. The results suggest an important unrecognized role for vagal tone in the regulation of cardiovascular function in mice and support the need for considerable caution when using mouse models of obesity to examine regulation of cardiovascular function. We argue that mouse physiology studies should be performed in thermoneutral conditions.

Cardiovascular and metabolic responses to fasting and thermoneutrality in Ay mice.

Several lines of evidence support a role for reduced melanocortin signaling in the regulation of metabolic rate and cardiovascular function during negative energy balance. We tested the hypothesis that agouti yellow (B6.Cg-A(y)) mice would exhibit blunted physiologic responses to fasting and thermoneutrality. Male B6.Cg-A(y) mice (A(y); n=11, 34+/-2 g) and lean B6 littermates (B6; n=7, 26+/-2 g) were implanted with telemetry devices and housed in metabolic chambers (T(a)=23 degrees C) to determine the effects of a 24-h fasting and exposure to thermoneutrality (T(a)=30 degrees C) on mean arterial pressure (MAP), heart rate (HR), AP and HR variability (time and frequency domain), oxygen consumption (VO(2)), and locomotor activity. A(y) mice exhibited elevated baseline light-period MAP (A(y): 113+/-4; B6: 99+/-3 mm Hg) and VO(2) (A(y): 1.82+/-0.08 vs. B6: 1.45+/-0.13 ml/min) with no difference in HR (A(y): 530+/-12 vs. B6: 548+/-19 bpm). At 12-24 h after food removal, A(y) mice displayed normal fasting-induced bradycardia (A(y): -106+/-12; B6: -117+/-19 bpm) and reduction in VO(2) (A(y): -0.19+/-0.04 vs. B6: -0.28+/-0.05 ml/min), but with augmented hypotension (A(y): -9+/-2 vs. B6: -0.5+/-2 mm Hg) and blunted hyperactivity (A(y): 27+/-23 vs. B6: 122+/-42 m/11 h). Fasting was associated with increased HR variability in both time and frequency domain in B6 but not A(y) mice. Exposure to thermoneutrality produced comparable reductions in MAP, HR, and VO(2) in both strains. We conclude that inhibition of melanocortin signaling is not requisite for, but participates in, the metabolic and cardiovascular responses to negative energy balance.

Cardiovascular responses to caloric restriction and thermoneutrality in C57BL/6J mice.

We utilized variations in caloric availability and ambient temperature (T(a)) to examine interrelationships between energy expenditure and cardiovascular function in mice. Male C57BL/6J mice (n = 6) were implanted with telemetry devices and housed in metabolic chambers for measurement of mean arterial pressure (MAP), heart rate (HR), O(2) consumption (VO(2)), and locomotor activity. Fasting (T(a) = 23 degrees C), initiated at the onset of the dark phase, resulted in large and transient depressions in MAP, HR, VO(2), and locomotor activity that occurred during hours 6-17, which suggests torporlike episodes. Food restriction (14 days, 60% of baseline intake) at T(a) = 23 degrees C resulted in progressive reductions in MAP and HR across days that were coupled with an increasing occurrence of episodic torporlike reductions in HR (<300 beats/min) and VO(2) (<1.0 ml/min). Exposure to thermoneutrality (T(a) = 30 degrees C, n = 6) reduced baseline light-period MAP (-14 +/- 2 mmHg) and HR (-184 +/- 12 beats/min). Caloric restriction at thermoneutrality produced further reductions in MAP and HR, but indications of torporlike episodes were absent. The results reveal that mice exhibit robust cardiovascular responses to both acute and chronic negative energy balance. Furthermore, we conclude that T(a) is a very important consideration when assessing cardiovascular function in mice.

Cardiovascular and metabolic responses of hypertensive and normotensive rats to one week of cold exposure.

Challenges to energy homeostasis, such as cold exposure, can have consequences for both metabolic and cardiovascular functioning. We hypothesized that 1-wk cold exposure (4 degrees C) would produce concurrent increases in metabolic rate (VO(2); indirect calorimetry), heart rate (HR), and mean arterial blood pressure (MAP) measured by telemetry. In the initial hours of change in ambient temperature (T(a)), both spontaneously hypertensive rats (SHRs) and normotensive Sprague-Dawley rats showed rapid increases (in cold) or decreases (in rewarming) of VO(2), HR, and MAP, although the initial changes in MAP and HR were more exaggerated in SHRs. Throughout cold exposure, HR, VO(2), food intake, and locomotor activity remained elevated but MAP decreased in both strains, particularly in the SHR. During rewarming, all measures normalized quickly in both strains except MAP, which fell below baseline (hypotension) for the first few days. The results indicate that variations of T(a) produce rapid changes in a suite of cardiovascular and behavioral responses that have many similarities in hypertensive and normotensive strains of rats. The findings are consistent with the general concept that the cardiovascular responses to cold exposure in rats are closely related to and perhaps a secondary consequence of the mechanisms responsible for increasing heat production.

Concurrent reductions in blood pressure and metabolic rate during fasting in the unrestrained SHR.

Fasting produces multiple cardiovascular, metabolic, and behavioral responses. To examine the interrelationship between these responses, male spontaneously hypertensive rats (SHR; n = 8) implanted with cardiovascular telemetry devices were housed in metabolic chambers at 23 degrees C for 22-h daily measurements of physiological variables. The experimental apparatus was designed so that ingestive behavior was detected by photobeams and locomotion was detected by a load sensor. Cardiovascular and metabolic status were determined as both a function of the circadian cycle (12-h dark and 10-h light), as well as during periods of inactivity (no ingestion and minimal locomotion) within the dark and light phases. Data were obtained during baseline, 48-h of caloric deprivation, and 6 days of refeeding. Fasting produced significant reductions in mean arterial pressure (dark: -9.2+/-1.3 from 143.7+/-3.7 mm Hg; light: -8.6+/-1.8 from 140.1+/-3.7 mm Hg), heart rate (dark: -43.4+/-5.2 from 330.0+/-5.2 beats/min; light: -27.4+/-5.2 from 294.0+/-5.2 beats/min), and oxygen consumption (dark: -5.0+/-0.6 from 20.6+/-0.3 ml x min(-1) x kg (0.75); light: -2.7+/-0.2 from 14.9 +/-0.2 ml x min(-1) x kg(0.75)). Analysis of inactive periods during both light and dark phases revealed that these reductions were not dependent on behavioral effects. We conclude that fasting produces concurrent and interrelated reductions in cardiovascular and metabolic function in the SHR. The merging of cardiovascular telemetry, indirect calorimetry, and behavioral monitoring provides a powerful approach for investigation of the integrative physiological responses to energetic challenges.

Shivering and digestion-related thermogenesis in pigeons during dark phase.

The pigeon's main source of regulated heat production, shivering, is especially likely to be used for thermoregulation during the dark phase of the day when there is little heat from locomotor activity. However, food stored in the pigeon's crop is digested during the night, and digestion-related thermogenesis (DRT) will provide heat that should decrease the need for shivering to maintain body temperature (Tb). We investigated the conditions under which DRT alters the occurrence of nocturnal shivering thermogenesis in pigeons. In fasting experiments, in which DRT was minimal, variations in pectoral shivering were closely related to the kinetics of nocturnal Tb when the ambient temperature (Ta) was moderate (21 degrees C). In that case, shivering was low while Tb fell at the beginning of the night, moderate during the nocturnal plateau in Tb, and strong during the prelight increase in Tb. Similar kinetics of nocturnal Tb occurred when Ta = 28 degrees C, but shivering was negligible throughout the dark phase. In restricted feeding experiments, nocturnal DRT was varied by providing different amounts of food late in the light phase. When Ta = 21 degrees C, 11 degrees C, and 1 degrees C, nocturnal Tb and O2 consumption were directly related to the amount of food ingested. However, nocturnal shivering tended to decrease as the food load increased and was significantly reduced at the higher loads. Because nocturnal shivering did not become more efficient in producing heat as the size of the food load increased, we conclude that nocturnal DRT decreased the need for shivering thermogenesis.

Reciprocity between light intensity and rhodopsin concentration across the rat retina.

1. If a purpose of photostasis - absorption of a constant number of photons by the retina, regardless of incident light levels - is to maintain rods at saturation during the light period, then in retinal regions where light intensity is low, rhodopsin concentration should be high, and vice versa. 2. Our ocular transmission photometric measurements revealed that the distribution of light intensity across the rat retina was not as simple as had been thought and, furthermore, that the local concentration of rhodopsin had a high negative correlation with the light intensity. 3. The reciprocity between these two parameters leads to nearly uniform rates of photon absorption in rods across the retina.

Shivering thermogenesis in the pigeon: the effects of activity, diurnal factors, and feeding state.

Shivering (electromyographic activity of the pectoral muscle), oxygen consumption, and body temperature were measured from undisturbed pigeons for periods of several weeks, and segments from the midparts of each phase of the light-dark cycle were compared at various ambient temperatures and feeding regimes. Behavior was recorded with a video camera. None of the observed types of behavior (e.g., walking, preening, feeding, drinking, pecking, defecation) induced spurious electrical activity in the pectoral muscle. On the other hand, none of these behaviors directly inhibited ongoing shivering. There was no difference in the mean level of shivering between the light (L) and dark (D) phases of the day in any of the conditions, although body temperature was 2 degreesC higher during L. Measurements of integrated electromyogram (EMG) with high temporal resolution (28 samples/s) showed that, at 1 degreesC, shivering in the pectoral muscle was present for more than 98% of the time. Plots of oxygen consumption against root mean square EMG were obtained in each condition by a filtering procedure that excludes data points in which oxygen consumption is affected by motor activity. These plots showed that the increase in heat production induced by a unit increase in pectoral EMG was lower in D than in L and that it was further lowered by fasting. The amplitude spectra of raw EMG signals were similar in all conditions. Spectra of demodulated (rectified, low-pass filtered) EMG showed a distinct rhythmicity around 8 Hz at 21 degreesC that was further enhanced by fasting but absent at 1 degreesC. This suggests that the degree of synchronization and pattern of recruitment of motor units are specific for various temperatures and feeding regimes, and may partly explain the variable relation between heat production and muscle electrical activity. The results emphasize the advantages of long-term measurements for understanding the control of thermogenesis in birds.

Vigilance states and body temperature during the circadian cycle in fed and fasted pigeons (Columba livia).

Fasting induces nocturnal hypothermia in pigeons. Slow-wave sleep (SWS) and paradoxical sleep (PS) are associated with reduced heat production in pigeons. The possibility that fasting-induced nocturnal hypothermia is related to increased SWS and PS was examined by comparing body temperature (Tb) and vigilance states when pigeons were fed and fasted. The results showed that when Tb is decreasing near the beginning of the dark phase, the percentage of total recording time (%TRT) spent in SWS and PS was elevated in fasting due to increased frequency of episodes and increased duration of PS episodes. When Tb was low during the middle segment of the dark phase, SWS was elevated in fasting due to increased episode duration. However, fasting did not alter PS, which increased in %TRT across the segment due to increased episode frequency. When Tb was rising during the final hours of dark, SWS remained elevated in fasting and %TRT in SWS and PS was relatively high. SWS and PS may promote the fasting pigeon's entry into, and maintenance of, nocturnal hypothermia.

Distribution of photon absorption rates across the rat retina.

1. An investigation into the distribution of light intensity across the rat retina was carried out on excised, intact rat eyes exposed to Ganzfeld illumination from a helium-neon laser (543 nm). 2. Some of the light entering the eyes exits through the sclera where its intensity can be monitored with an optical 'pick-up' that samples the intensity coming from a small region of external sclera and underlying retina. The spatial resolution of the pick-up is such that it samples light that has passed through ca 2 % of the rods in the rat eye. 3. Some of the laser light is absorbed by the rod pigment, rhodopsin, which gradually bleaches. Bleaching in the retina, in turn, causes an exponential increase in intensity emanating from the sclera. By monitoring this intensity increase, we are able to measure two important parameters in a single bleaching run: the local rhodopsin concentration and the local intensity falling on the rods. 4. With an ocular transmission photometer, we have measured both the local intensity and the local rhodopsin concentration across wide regions of rat retina. Both pigmented and albino rats were studied. 5. The distributions of rhodopsin and intensity were both nearly uniform; consequently, the product, (rhodopsin concentration) x (intensity), was similarly nearly equal across the retina. This means that the initial rate of photon absorption is about the same at all retinal locations. 6. Interpreted in terms of photostasis (the regulation of daily photon catch), this means that the rate of photon absorption is about the same in each rod, viz. 14 400 photons absorbed per rod per second. Since this rate of absorption is sufficient to saturate the rod, one possible purpose of photostasis is to maintain the rod system in a saturated state during daylight hours.

Prolongation of the postprandial intermeal interval by gastrin-releasing peptide1-27 in spontaneously feeding rats.

The present study explored the effects of intravenous gastrin-releasing peptide1-27 (GRP) on the postprandial intermeal interval (IMI) when delivered shortly after termination of the first spontaneous nocturnal meal in freely-feeding rats. Undisturbed, ad lib-fed (milk), male rats (n = 11) with chronic inferior vena caval catheters were infused with saline and each of three doses (2.5, 5 and 10 nmol/kg) of GRP in counterbalanced order. Infusions began 5 min after the last lick of the first nocturnal meal and continued for 2 min (60 microliters/min), with delivery of the peptide during the first minute. Infusions and recording of meal data (licks) were fully automated and computer-controlled. Both 5 and 10 nmol/kg of GRP significantly prolonged the IMI by over 50%, but had no effect on the size of the following meal. This is the first demonstration of the prolongation of the IMI by intravenous GRP in undisturbed, freely-feeding rats, and the result suggests that endogenous GRP may play a role in the control of the postprandial intermeal interval.

High lick rate is maintained throughout spontaneous liquid meals in freely feeding rats.

To investigate the microstructure of spontaneous meals in freely feeding rats, 16 adult male Sprague Dawley rats were housed individually in custom-designed lickometer cages and maintained on a milk diet. Licks were recorded over 23 h at millisecond accuracy via a computer-controlled lickometer. Analysis of lick data revealed an average of about 12 discrete meals/day occurring mainly during the dark phase. The most striking feature of both dark and light meals was the maintenance of a high initial rate of licking until an abrupt decline at the end of the meal. This pattern of licking is very different from the exponential decay of lick rate reported in scheduled test meals of palatable solutions. Thus, the microstructure of licking for meals is affected in an apparently fundamental way by whether a meal is scheduled or spontaneous, suggesting a basic difference in the underlying physiologic controls.

Nocturnal digestion, cloacal excretion, and digestion-related thermogenesis in pigeons (Columba livia).

Two indicators of nocturnal digestive activity were identified in pigeons. Experiment 1 showed that a sizable amount of food empties from the crop while pigeons are inactive during the night. Experiment 2 showed that the number and volume of cloacal droppings during the night were directly related to the volume of food consumed during the day. The temporal pattern of cloacal droppings in the night was systematically related to features of the nocturnal body temperature (Tb) curves, suggesting that excretory activity is thermogenic in its own right and/or that it is a marker for a thermogenic process in the upper digestive tract. Questions about the relationship between digestion-related thermogenesis and shivering thermogenesis during the night in birds are highlighted by these findings.

Daily cycles in body temperature, metabolic rate, and substrate utilization in pigeons: influence of amount and timing of food consumption.

Pigeons lived in individual chambers where instantaneous metabolic rate (MR; indirect calorimetry), body temperature (Tb), and substrate utilization (RQ) were measured 24 times each hour throughout the 12h:12h light:dark cycle. The amount of food consumed influenced the amplitude of the MR and Tb cycles, primarily by affecting the dark-phase segment of the cycle: when food was consumed ad lib, low-amplitude daily cycles in MR and Tb occurred in which levels in the dark phase were lower than in the light; during reduced food intake in restricted feeding or in fasting, high-amplitude cycles occurred primarily because nocturnal hypometabolism and hypothermia developed; in restricted feeding, the level of MR and Tb during the dark-phase segment of the cycle was directly related to short-term variation in amount consumed. The timing of food consumption primarily affected the light-phase segment of the MR and Tb cycles: when feeding was restricted to a time late in the light phase, these measures became depressed early in the light phase, and then greatly elevated near the scheduled time of feeding. This distinctive light-phase pattern developed quickly after the restricted feeding schedule began and may reflect the influence of a circadian food-entrainable oscillator. RQ indicated carbohydrate utilization for most of the 24-h cycle during ad lib feeding and in restricted feeding. However, approximately 2 h before the first feeding bout of the day, the RQ cycle indicated a sizable shift towards lipid utilization, which terminated after the bout was completed. There was a smaller, more transient, decrease in RQ near the time of the light-dark transition, which may imply cessation of digestive activity in preparation for the nocturnal decrease in Tb. During fasting, RQ indicated lipid utilization throughout the entire cycle. Whole-day energy expenditure by pigeons in these laboratory circumstances was shown to be closely related to the changes in within-day cycles associated with variations in the amount and timing of food intake.

Ingestive behavior and body temperature of pigeons during long-term cold exposure.

Pigeons well adapted to living in 21 degrees C ambient temperature (T(a)) were continuously exposed to cold T(a) for 6 days (Experiment 1) or 30 days (Experiment 2). The pigeons lived on a 12:12 L:D cycle where they could obtain short access to food and water at any time in the light phase by making 12 keypeck responses. Across the two experiments, the levels of cold exposure were 6 degrees C, 1 degrees C, and 0.6 degrees C. In cold, total daily food intake increased gradually across the first few days of the exposure, and the new level of food intake was inversely related to T(a). The daily ratio of total water drunk to total food eaten averaged approximately 1.3 in 21 degrees C, fell to approximately 1.0 on day 1 of cold exposure, and remained at that level for as long as 30 days. The day-night cycle in core body temperature was not changed by cold exposure. The bimodal temporal pattern of feeding in the light phase, which is characteristic of pigeons in moderate T(a), was preserved in the cold although the absolute level of feeding activity was enhanced. An analysis of the ways individual pigeons achieved total daily food intake by combining a number of feeding episodes with an average amount eaten per episode revealed considerable variation between birds, but a relatively constant feeding style within bird over time. In the colder temperatures used, however, the pigeons all increased the number of feeding episodes per day. The results provide the first detailed analysis of cold-induced changes in ingestive behavior in the pigeon and raise several questions about the behavioral expression of cold-sensitive regulatory processes.

Energetic responses of pigeons during food deprivation and restricted feeding.

Pigeons ate food ad lib, then fasted for several days, and finally ate a controlled amount of food once a day for several months to maintain body weight at 80% of the ad lib value. Whole-body dry heat loss (HL) and core body temperature (Tb) were measured continuously for each pigeon. Thermal conductance (C) was calculated from HL and Tb. Relative to ad lib feeding, 24-h HL was reduced by approximately 50% during fasting and controlled feeding. The majority of energy savings was achieved by lowering C, which appeared to maintain a saturated low value throughout most of the light and dark phases. Therefore, the pigeon's characteristic high Tb in the daily light phase during fasting and food restriction does not necessarily imply high energy expenditure. In the dark phase, the fasting pigeon's characteristically low Tb enhances energy savings already being achieved through lowered C. The daily cycle in Tb, and to a lesser extent in HL and C, was strongly altered by occasional probe variations in the amount of food given at the single daily feeding and by a shift in the time of the daily feeding.

Overview of infectious diseases, Part 2.

The choice of drug therapy in the treatment of infectious disease depends on many factors, including the probable sensitivity of likely pathogens and previous efficacy of the agent. Pharmacokinetic factors are crucial since good in vitro results cannot predict in vivo success unless the drug is present in effective concentration at the infection site. Combination therapy can be used in certain situations to improve the chances of clinical success. Part 3 of this series will conclude the discussion of drug therapy. The general manifestations of and recovery process from infectious diseases will also be discussed.