Counteraction of spaceflight-induced changes in the rat central serotonergic system by adrenalectomy and corticosteroid replacement.

The effects of a 17-day spaceflight duration on serotonergic measures in various parts of rat brain have been studied (flight-SHAM group). The contribution of the activation of the hypothalamo-pituitary-adrenal axis (HPA) related to the response of the central serotonin system was evaluated in adrenalectomized with chronic corticosterone replacement rats (flight-ADX+CORT group). These two groups of rats were compared to their respective ground-based controls. Physiological parameters (body, adrenal and thymus weights) and corticosterone levels were measured. In flight-SHAM group as compared to controls, adrenal hypertrophy and elevation in plasma corticosterone levels (174%) were observed, without change in thymus mass. In most brain areas studied, significant decreases in TRP, 5-HTP and 5-HIAA were found associated with lower levels of 5-HT in cortex, thalamus and striatum. Conversely, there were elevations in TRP, 5-HTP levels in striatum and increases in 5-HIAA/5-HT ratios, an index of 5-HT turnover, in cortex, striatum and olfactory bulb while the hypothalamus was the sole region where a fall was observed. In ADX rats with chronic corticosterone replacement these effects were not observed in the majority of brain areas. It is concluded that a 17-day spaceflight exerted an inhibitory effect on serotonin metabolism, probably by activation of the HPA axis. The results could not distinguish between the effects of microgravity and the stress associated with landing.

Central and peripheral sympathetic activities in rats during recovery from simulated weightlessness.

Rats were tail suspended, keeping their forelimbs weight bearing for 14 days, and then allowed to recover for a short (6-h) or a long (24-h) period to assess the behavior of the sympathetic nervous system after weightless simulation. Sympathetic activity was determined by measuring norepinephrine (NE) turnover in the brain stem cell groups involved in central blood pressure control and in organs playing a key role in the cardiovascular regulation (heart and kidneys). The NE turnover was greatly reduced in the rostral (-56%; P < 0.001) and caudal (-73%; P < 0.001) A2 nucleus of suspended rats but was unchanged in the A1, A5, and A6 cell groups compared with attached rats. The NE turnover in the cardiac atria (-34%; P < 0.001) and ventricles (-35%; P < 0.001) and kidneys (-31%; P < 0.001) was decreased after suspension. The central and peripheral sympathetic activities returned to normal within 24 h of release from suspension, but there was hyperactivity after 6 h of recovery. This raises the problem of interpreting the results obtained in animals killed a few hours after return from spaceflight.

Restraint vs. hindlimb suspension on fluid and electrolyte balance in rats.

To determine the effect of hindlimb suspension on body fluid volume, salt and water balance, and relevant hormones, two series of experiments were performed in an experimental protocol including periods of isolation (7 days), horizontal attachment (7 days), and suspension (14 days). 1) During the first experiment, water and electrolyte balance, arginine vasopressin (AVP), and guanosine 3',5'- cyclic monophosphate (cGMP) were determined in urine, atrial natriuretic peptide in plasma and atria, and renin concentration and AVP in plasma in 30 rats. 2) During the second experiment, blood volume and extracellular fluid volume were measured by a dilution technique (Evans blue and sodium thiocyanate) in another 30 rats. We observed a pronounced and early effect of horizontal attachment on the renal variables. After 48 h, diuresis (49%), natriuresis (44%), kaliuresis (36%), osmotic load (39%), creatinine (28%), and AVP excretion (155%) were significantly increased in attached rats (P < 0.05). There was no short-term (24-h) effect of suspension on urine flow and Na+, K+, creatinine, and AVP excretion, but the urine cGMP decreased significantly (45%; P < 0.05). Significant decreases in natriuresis, kaliuresis, urine creatinine, and osmotic load occurred in the suspension group 7 days after suspension. After the 14-day tail suspension, plasma volume and extracellular fluid volume measured in suspended rats were not different from isolated rat values, whereas plasma volume increased by 15% (P < 0.05) in the attached rats. Plasma immunoreactive plasma atrial natriuretic levels of suspended rats were significantly reduced by 35% vs. isolated rats (P < 0.001) and by 18% vs. attached rats (P < 0.05). By using this experimental protocol, the physiological alterations revealed that suspension produced some acute and long-term effects, but the fixation to the suspension device, restraint, and confinement have their own influence on fluid distribution and renal function.

Recording heart rate and blood pressure in rats during parabolic flight.

The way in which the cardiovascular system adapts to weightlessness is still under discussion. No data are yet available on the responses of rats during space flight, although this animal is commonly used in simulation studies. We have designed and tested a protocol to study the short term responses of the cardiovascular system to weightlessness during parabolic flight. A telemetry system was used to measure heart rate (HR) and blood pressure. It was possible to collect and record radio-signals without any interference. Microgravity caused a reduction in HR, an increase in mean arterial pressure (MAP, 7%), and a non-significant decrease in central venous pressure (CVP, 13%). The change in CVP was similar to the decrease observed in human space flight. This type of study may also be feasible for longer exposure of rats to microgravity (space flight).

Tissue norepinephrine turnover and cardiovascular responses during intermittent dehydration in the rat.

We investigated the central and peripheral sympathetic responses to intermittent dehydration in rats. The norepinephrine (NE) turnover, a biochemical index correlated with noradrenergic neuronal activity, was measured. The modification of blood pressure was also determined by telemetry during the different cycles of dehydration. Dehydration caused a decrease of NE turnover in A2, A5 and A6 nuclei and in peripheral organs. The vasopressinergic level of dehydrated rats decreased in hypophysis and hypothalamus, and increased in plasma. A repeated gradual increase of arterial blood pressure during the first three days of dehydration, followed by a sudden drop when the rats were rehydrated on the fourth day was observed. In conclusion, our study revealed an increase in blood pressure and in central sympathetic activity during dehydration.

Effect of acute atenolol on short-term blood pressure variability and baroreflex sensitivity in rats.

Beta-blocker therapy for hypertension or coronary artery disease is common, but there are a lot of controversies about its effects on short-term blood pressure variability and arterial baroreceptor reflexes. The aim of this study was to evaluate the effects of acute atenolol on baroreflex sensitivity (BRS) and on the spontaneous variability of systolic blood pressure (SBP) and RR intervals in conscious rats. Ten Wistar rats equipped with telemetry system were evaluated: 1) under control conditions; 2) after injection of saline; and 3) during beta1-adrenergic blockade by atenolol. Fast Fourier transform analysis was applied to RR intervals and SBP. Atenolol increased RR intervals significantly by 14% and the variation coefficient of the RR intervals by 31%. SBP was reduced significantly by 9%. In frequency domain, beta1-blockade in RR intervals increased very low frequency by 33% and the total power by 22% and decreased low frequency by 25%. The ratio of low to high frequency power decreased by 60%. Frequency domain variables in SBP were not significantly changed after beta1-adrenergic receptor blockade. BRS (gain alpha) was not significantly altered by beta-blockers. Acute atenolol decreased SBP and increased RR intervals with no change in BRS, indicating 'resetting' of baroreflex function.

Smoothed pseudo Wigner-Ville distribution as an alternative to Fourier transform in rats.

Techniques for examining signals in the time and frequency domains are well-established tools. These tools have their limitations; they tell us in a broad sense where the signal component exists in the frequency domain, but they do not tell us how its frequency characteristics change over time. The time-frequency has become a powerful alternative for the analysis of signals. Among various time-frequency distribution methods, one of the most studied is the Wigner-Ville distribution. The aim of this study was to evaluate in conscious rats smoothed pseudo Wigner--Ville distribution (SPWVD) as an alternative to the fast Fourier transform (FFT) in RR intervals and in systolic blood pressure (SBP), before and after adrenergic and cholinergic receptor blockade. Fourteen Wistar rats equipped with telemetry probe were evaluated: (1) under control conditions; (2) after injection of saline (100 microl kg(-1) i.v.); (3) after atenolol (1 mg kg(-1) i.v.); (4) after atropine methyl nitrate (0.5 mg kg(-1) i.v.); and (5) after phentolamine (5 mg kg(-1) i.v.). FFT and SPWVD were applied to RR intervals and SBP time series. Six-minute time series of RR intervals, systolic and diastolic pressures were analysed. The bias and distribution of differences between FFT and SPWVD methods in RR intervals under base conditions were 1.4+/-0.4% (r2=0.94; P<0.01) in LF/LF+HF: 1.5+/-0.5% ( r2=0.92; P<0.01) in HF/LF+HF and 4.8+/-1.9% (r2=0.92; P<0.01) in LF/HF. In SBP the bias and distribution were 1.5+/-0.8% (r2=0.90) P<0.05) in LF/LF+HF and 1.7+/-0.6% (r2=-0.92; P<0.01) in HF/LF+HF. In the frequency domain analysis of RR intervals and SBP there was no difference between FFT and SPWVD. The agreement between the methods demonstrates that in stationary signals both methods can be used interchangeably. SPWVD may be an interesting tool to analyse biomedical signals; it provides a good resolution at high frequency and a good frequency resolution at low frequencies independently if signals remain stationary.

Effect of a 14-day hindlimb suspension on beta-adrenoreceptors in rats.

Exposure to long-term simulated microgravity exhibits reduced sympathetic nervous system activity. This study tested the hypothesis that the hypersensitivity of adrenoreceptors would explain partly many other features of the hemodynamic consequences of return from space. The biochemical properties of the beta adrenoreceptors (betaAR) were determined using 125I-cyanopindolol (125I-CYP) binding in three rat groups: (1) The first experimental group consisted of 24 h-restrained orthostatic rats in the horizontal position, to test the early effect of the attachment to the suspension device; (2) the second experimental group consisted of 24 h-restrained antiorthostatic rats, to test the early effect of the suspension; (3) the third experimental group consisted of 14 day-restrained antiorthostatic rats, to test the long term effect of the suspension. The study was performed in two organs involved in blood pressure regulation, i.e. the heart (atria and ventricles were separated) and kidneys. The Scatchard analysis of 125I-cyanopindolol binding in both organs indicated no significant alterations in the dissociation constant (Kd) and the maximum binding capacity (Bmax) in the three experimental groups. These results do not allow the conclusion about the SNS adaptation pattern to simulated microgravity. Thus, the hypothesis that betaAR are involved in the cardiovascular adaptation to simulated microgravity is not verified in this model where, as a matter of fact, cardiovascular deconditioning is not verified even if this model is widely used.

Central and peripheral noradrenergic responses to 14 days of spaceflight (SLS-2) or hindlimb suspension in rats.

INTRODUCTION: The purposes of this work were to assess the influence of microgravity on the central and peripheral noradrenergic activity to reevaluate SLS-1 mission findings and to compare it with that of simulated microgravity in rats. METHODS: The norepinephrine (NE) contents of the brainstem cell groups (A1, A2, A5, and A6) and organs (heart and kidneys) involved in blood pressure regulation were determined in rats after a 14-d spaceflight (SLS-2 with animals sacrificed 6 h after landing) and after a 14-d hindlimb suspension followed with 6 h of recovery. RESULTS: After SLS-2 spaceflight, NE contents were not significantly different between flight and ground-based rats either in A1 (5.2 +/- 0.5 vs. 5.7 +/- 0.4 pmol/structure), rostral A2 (12.1 +/- 0.5 vs. 11.1 +/- 0.9 pmol/structure), caudal A2 (3.2 +/- 0.6 vs. 4.3 +/- 0.5 pmol/structure) and A5 (4.4 +/- 0.4 vs. 4.3 +/- 0.5 pmol/structure) nuclei or in cardiac atria (98.6 +/- 7.5 vs. 83.4 +/- 8.9 pmol.mg-1 protein), ventricles (38.3 +/- 2.2 vs. 44.1 +/- 2.8 pmol.mg-1 protein) and kidneys (13.4 +/- 0.8 vs. 17.7 +/- 1.5 pmol.mg-1 protein). NE content was unchanged in A6 nucleus after SLS-2 comparing with control rats (respectively 4.1 +/- 0.3 vs. 4.5 +/- 0.5 pmol/structure), while it was depleted after SLS-1 mission (2.9 +/- 0.3 vs. 8.8 +/- 0.7 pmol/structure, p < 0.001) probably in relation with the stressful conditions on return to Earth. Similarly, no alterations between suspended and control rats were noted in central and peripheral NE contents after 14 d of suspension and after 6 h of recovery, whereas NE turnover studies evidenced large changes in the activities on structures on suspension and on recovery. CONCLUSION: These results suggest that only NE turnover determination will provide information about the role of the sympathetic system in the cardiovascular deconditioning. This raises the problem of the necessity to experiment inflight (injections, sacrifice) in order to avoid the recovery effects of the few hours following the landing.

Biochemical characteristics of beta-adrenoceptors in rats after an 18-day spaceflight (LMS-STS78).

BACKGROUND: To ascertain whether there was autonomic adaptation with the development of adrenoceptor hypersensitivity under microgravity, the biochemical properties of the beta-adrenoceptors were determined using (125I)iodocyanopindolol (ICYP) binding in rats flown for 18 d onboard the space shuttle. METHODS: This study was performed on heart and kidneys of 3 groups of 12 animals: the flight and 2 ground control (vivarium and AEM) groups. To distinguish the possible role of the corticosteroids, half of each animal group was bilaterally adrenalectomized (ADX rats) with an aldosterone and corticosterone supplementation while the other half was SHAM operated. RESULTS: The Scatchard analysis of the ICYP-binding in both organs revealed no significant alterations in the dissociation constant (Kd) and in the maximal binding capacity (Bmax) between SHAM flight and control groups. The Kd of the beta-adrenoceptors in the cardiac atria of the SHAM flight rats (74 +/- 5 pm) was significantly higher (p < 0.05) than in those of the ADX flight rats (60 +/- 3 pm) while the Bmax was nonsignificantly higher (1925 +/- 370 in SHAM flight rats vs. 1482 +/- 283 fmol x mg(-1) protein in ADX flight rats). No significant change was determined for the Bmax and Kd values in the kidneys of the ADX and SHAM flight rats. CONCLUSIONS: This work performed on animals did not show any obvious effect of microgravity on the beta-adrenergic function in the heart and kidneys. Inflight rodent sacrifice protocols should definitely ensure assessment of the influence of microgravity on the animals.

Are energy metabolism alterations involved in cardiovascular deconditioning after weightlessness? An hypothesis.

The physiopathogenesis of the cardiovascular deconditioning syndrome observed after actual and simulated microgravity is still under debate, despite numerous studies conducted on the role of blood volume, hormones involved in its regulation, sympathetic nervous system, baroreflexes and venous compliance. Orthostatic intolerance, a reduced exercise capacity and an increased heart rate at rest characterize this syndrome. Recent data suggest, first, the presence of a complex loop between the sympathetic nervous system, carbohydrate metabolism (insulin) and leptin hormone and, second, that this loop, an overall reflection of energy metabolism, participates in cardiovascular regulation. After a resume of studies conducted on fuel homeostasis during actual and simulated microgravity, the possible implications of energy metabolism in the development of the cardiovascular deconditioning syndrome will be discussed.

Assessment of autonomic cardiovascular indices in non-stationary data in rats.

The analysis of heart rate in the frequency domain has become increasingly important in physiological studies, and supports the use of heart rate variability as an index of autonomic cardiovascular control. A new index, the instant centre frequency (ICF) has been proposed as a global index of the instantaneous relationship between sympathetic and vagal modulation. The aim of this study was to assess ICF, RR intervals, and heart rate variability measures as indices of sympathovagal balance during a pharmacological blockade of the autonomic nervous system in normotensive rats. RR intervals and arterial blood pressure of 10 conscious Wistar rats equipped with telemetry probes, were evaluated before, during, and after injection of: (1) saline (100 microl kg(-1) i.v.); (2) phentolamine (5 mg kg(-1) i.v.); (3) atropine methyl nitrate (0.5 mg kg(-1) i.v.); and (4) atenolol (1 mg kg(-1) i.v.). RR interval series were analysed by the smoothed pseudo-Wigner-Ville distribution. A general linearised model was used to evaluate the parameters. ICF was calculated in the same way as the peak power frequency by use of the first moment of instant spectrum. We calculated the ICF of the whole spectrum (ICF(T)), ICF in high frequency (ICF(H)) and ICF in low frequency (ICF(L)). The RR intervals and ICF indexes varied similarly and presented the lowest coefficient of variation among animals exposed to the same autonomic conditions. ICF(T)-ICF(L) and ICF(H)-ICF(T) were strongly correlated with normalised HF and normalised LF. In normotensive rats, RR intervals and ICF indices may reliably capture the effects of the sympathetic and parasympathetic nervous system on the sinus node.

Vasopressin and A1 noradrenaline turnover during food or water deprivation in the rat.

In the present study, we have examined in Wistar rats the effects of food or water deprivation of 3 days on the hypophyso-adrenal axis, vasopressinergic system and activity of A1 noradrenergic brain stem cell group, which is involved in the control of the hypothalamic neuro-endocrine activity. Levels of adrenocorticotropic hormone (ACTH) and vasopressin (AVP) were determined by radio-immunoassay, and corticosterone level was determined by fluorimetric method. Plasma levels of ACTH and corticosterone were greatly increased in both groups of rats. In water-deprived rats, plasma AVP (13.83 +/- 1.63 vs. 3.03 +/- 0.23 pg/ml) and osmolality levels were significantly elevated with a marked decrease of AVP hypophysis content (272 +/- 65 vs. 1098 +/- 75 ng/mg protein), but not in food-deprived rats in which osmolality did not change and AVP remained stocked (2082 +/- 216 ng/mg protein) in the hypophysis without release in the plasma (1.11 +/- 0.23 pg/ml). These observations indicated that both food-deprivation and water-deprivation stimulated the pituitary adrenal axis thereby suggesting a stress state. AVP production is stimulated both by fluid and food restriction but is secreted with differential effects: during food restriction AVP secretion is limited to supporting the hypothalamic pituitary-adrenal system.

Autonomic control of cardiovascular dynamics during weightlessness.

Measuring cardiovascular dynamics is a new method of assessing the autonomic regulation of the cardiovascular system, it provides an easily-implemented non-invasive way of monitoring the effects of weightlessness on this regulatory function. The major findings of studies on cardiovascular dynamics during actual or simulated weightlessness are presented, taking into account the recent consensus on this approach. Future improvements of these studies are discussed.

Blood pressure and heart rate determination by radiotelemetry in mice.

Mice are not commonly used in cardiovascular physiology, especially in space physiology because of methodological problems. The initial studies that have monitored arterial pressure and heart rate in mice used the tail cuff plethysmography method or classical catheterization techniques with a catheter liable to an external pressure transducer. But for long term arterial pressure measurements the studies have been facilitated by the development of radiotelemetry. This is a less constraining method as classical one allowing to monitor in continuous in freely moving animal blood pressure, heart rate and temperature. This technique allows to record these parameters thank to an implanted transmitter without physical connection with the monitoring system. The aim of this work was to valid the use of the radiotelemetry in mice to evaluate arterial blood pressure and heart rate during different stress conditions (but only control data are described in this paper).

Effects of acute and chronic starvation on central and peripheral noradrenaline turnover, blood pressure and heart rate in the rat.

When faced with stress, an organism calls upon several mechanisms to maintain biological homeostasis. The cardiovascular system is the first to respond usually with an increase in arterial pressure and tachycardia. Therefore we investigated the central and peripheral sympathetic responses to acute and chronic starvation in Wistar rats. The noradrenaline (NA) turnover rate was determined in different catecholaminergic nuclei (A1, A2, A5, A6) as well as the arterial blood pressure and heart rate modifications. During acute starvation (3 days of starvation), the NA turnover was increased in the A1 and rostral A2 nuclei as well as in ventricles and kidneys and decreased in the A6 nucleus. During chronic starvation (4 consecutive cycles of 3 days of starvation plus 1 day of feeding), the NA turnover was increased in the A5 and caudal A2 nuclei as well as in ventricles and atria and decreased in the A1 nucleus and kidneys. The arterial blood pressure revealed a gradual decrease during the first 3 days of fasting but the heart rate was not modified. We conclude that starvation should be considered as an unusual state of stress because of the absence of locus coeruleus response (A6 nucleus) despite its well-defined role in stress reactions. One of the manifestations of these central and peripheral noradrenergic changes is the change in blood pressure during the starvation-feeding cycles.

Regional blood flow in conscious rats after head-down suspension.

Exposure to microgravity in humans causes cardiovascular deconditioning affecting blood pressure, heart rate and vascular responsiveness. This study investigated cardiac output, arterial blood pressure and regional blood flows [radioactive microspheres: 57Co, 15.5 (SEM 0.1) microm in diameter] in conscious and freely moving rats subjected to 14 days of simulated microgravity (head-down suspension, HDS) in male Wistar rats: control (horizontally attached, n = 7), suspended for 14 days (n = 8) and suspended/allowed to recover for 10 min (R10min, n = 5) or 24 h (n = 9). Compared to the control group, 14 days of HDS resulted in reduced total peripheral resistance (37%); an increased cardiac index (65%) was associated with no significant change in the mean arterial pressure BPa. There were elevated brain (63%), visceral (> 20%), hindlimb (> 80%) and forelimb (> 215%) muscle blood flows. In the R10min group, the BPa decreased (18%) and the regional blood flows returned to control values. Within 24 h the BPa as well as cardiac index and total peripheral resistance were restored. In conclusion, 14 days of HDS engendered local circulatory changes resulting in transient blood pressure instability during recovery.

Simulated weightlessness alters the nycthemeral distribution of energy expenditure in rats.

The energy metabolism adaptations to simulated weightlessness in rats by hindlimb tail suspension are unknown. 12 male rats were assigned to 7 days of isolation, 7 days of habituation to the suspension device, 10 days of simulated weightlessness, and 3 days of recovery. The 24-hour energy expenditure was measured by continuous indirect calorimetry. We calculated the 12-hour energy expenditure during the active (night) and inactive (day) periods, the minimal observed metabolic rates with the day values taken as an index of the basal metabolic rate, and the non-basal energy expenditure representing the cost of physical activity plus the diet-induced thermogenesis. Suspension did not change the mean 24-hour energy expenditure (360.8+/-15.3 J min(-1) kg(-0.67)), but reduced the night/day difference by 64 % (P<0.05) through a concomitant drop in night-energy expenditure and increase in day values. The difference between night and day minimal metabolic rates was reduced by 81 % (P<0.05), and the transient rise in day values suggests an early and moderate basal metabolic rate increase (9 %). An overall 19 % reduction in non-basal energy expenditure was observed during simulated weightlessness (P<0.05), which was mainly attributable to a reduction in the cost of physical activity. 3 days of recovery restored the night/day differences but increased the 24-hour energy expenditure by 10 % (P<0.05). In conclusion, hindlimb tail suspension in rats did not alter the 24-hour energy expenditure, but it transiently increased the basal metabolic rate, and altered both the energy expended on physical activity and the nycthemeral distribution of motor activity. These data suggest that the circadian rhythms of energy expenditure are affected during simulated weightlessness.

Cardiovascular adaptations to parabolic flight in rats: a radio-telemetry feasibility study.

This experiment was a feasibility study which consisted in investigating arterial blood pressure and heart rate to transient and repeated exposure to microgravity in eight unrestrained rats previously implanted with radio-telemetry transmitter. The aim was to perform such recordings throughout all the phases of a parabola during parabolic flights. This study revealed that it was possible to collect the radio-signal without any interference with electronic or magnetic environment. We observed in microgravity a significant reduction in heart rate (6%) and a significant increase in arterial blood pressure (7%). In conclusion, such a study seems to be feasible during longer exposure to microgravity (space flight) in order to study the cardiovascular adaptation in rat.