Presystemic signals in the control of thirst, salt appetite, and vasopressin secretion

Presystemic signals play an important role in the control of ingestive behavior by allowing animals to anticipate imminent physiological changes. The significance of such signals in the control of food intake has been amply demonstrated and is widely appreciated. Our recent experiments have revealed that presystemic signals also provide important early feedback when rats drink water or NaCl solution, before the ingested fluids are absorbed and influence cerebral osmoreceptors or cardiovascular baroreceptors. These early signals clearly affect vasopressin (VP) secretion and thirst. They relate either to the distension of the stomach and proximal small intestine (presumably mediated by local stretch receptors) or to the concentration of fluid that empties from the stomach into the small intestine (presumably mediated by visceral osmo- or Na(+)-receptors). Dehydrated dogs use functionally comparable signals from the oropharynx while drinking in order to inhibit both VP secretion and thirst. However, that system differs in several respects from the system in rats aside from the fact that the presystemic signals in rats are not oropharyngeal: in rodents, (a) separate early signals influence VP secretion and thirst, (b) early signals can provide both stimulation and inhibition of VP secretion and thirst, and (c) the early signals are associated with both the volume and concentration of ingested fluid. These presystemic signals also inhibit the intake of NaCl solution by rats with salt appetite.     

Early osmoregulatory signals in the control of water intake and neurohypophyseal hormone secretion

Dehydrated dogs inhibit secretion of vasopressin (VP) within minutes after drinking water, before plasma osmolality (pOsm) diminishes. In recent studies, we found that water ingestion by rats similarly inhibits VP and oxytocin (OT) secretion rapidly, before pOsm is diluted. Adult male rats were infused with 1 M NaCl (2 ml/h iv) for 240 min to stimulate VP and OT secretion. After 220 min of infusion, rats were given water or isotonic saline (IS) to drink for 5 min, and blood samples were taken 5 and 15 min later. Plasma levels of VP (pVP) and OT (pOT) were much lower when rats ingested water instead of IS, even though rats drank comparable amounts of both fluids ( approximately 5.5 ml) and pOsm was not significantly affected in either case. In another study, rats were infused with 1 M NaCl (2 ml/h iv) for 120 min before receiving 4-ml gastric loads of either 0.5 M NaCl (HS) or IS; blood samples taken 25 min later showed that pVP and pOT were much higher when rats were given gastric loads of HS instead of IS, even though pOsm was not significantly altered. Comparable results were obtained when gastric loads of HS or IS were given to rats that had been deprived of drinking water overnight. Other dehydrated rats treated similarly but given access to drinking water consumed much more when they had been given gastric loads of HS instead of IS. Collectively, these and other findings suggest the importance of early signals, perhaps from hepatoportal osmoreceptors or Na(+) receptors, in the control of VP and OT secretion and water intake in rats.     

Hypotension and thirst in rats after phentolamine treatment

Administration of the α-adrenergic receptor blocker phentolamine is known to lower arterial blood pressure and to increase renin secretion and water intake in rats. In the present experiments, drinking by rats after subcutaneous administration of the drug was found to be inversely related to arterial blood pressure within the range of 60–90 mm Hg. Drug treatment in nephrectomized rats led to such severe hypotension that drinking was precluded. Pretreatment with propranolol moderated the hypotension in nephrectomized rats and drinking was not attenuated. These results parallel those of previous experiments using the β-adrenergic receptor agonist isoproterenol. Collectively, they suggest that after treatment with these hypotensive agents, a stimulus for thirst can arise from some factor other than angiotensin. This stimulus, perhaps mediated by arterial baroreceptors, is additive in its effects on thirst with the stimulus induced by hypertonic NaCl or subcutaneous colloid treatments.     

Neurohypophysial responses to emotional stress after deafferentation of sino-aortic baroreceptors in rats.

In an attempt to test the possibility that sino-aortic baroreceptors may mediate the previously reported stress response in hypothalamic magnocellular neurosecretory cell activity in rats, effects of deafferentation of sino-aortic baroreceptors on plasma levels of vasopressin and oxytocin after fear-related emotional stress were studied in male rats 28-33 days after the surgery. An alpha 1-adrenergic receptor agonist, phenylephrine (1 mg/kg) injected i.p. under anesthesia increased arterial blood pressure in the rats that had received surgical operation of sino-aortic denervation (SAD) and in the rats of sham-operation control (SHAM). Reflex bradycardia after phenylephrine occurred in the SHAM but not in the SAD group. These results indicate that afferent signals originating from sino-aortic baroreceptors were effectively blocked by the SAD surgery. In the similarly prepared SAD group, plasma level of vasopressin was decreased and plasma level of oxytocin was increased significantly to the same extent as in the SHAM group after low-frequency shocks (0.05 Hz, 5 min) or environmental cue signals previously paired with shocks. It is therefore suggested that afferent neural signals originating from sino-aortic baroreceptors are not primarily involved in the suppressive vasopressin or the facilitatory oxytocin response to fear-related emotional stress in rats.     

Hypovolemia stimulates intraoral intake of water and NaCl solution in intact rats but not in chronic decerebrate rats

This experiment tested the hypothesis that afferent signals from cardiac baroreceptors to the caudal brain stem are integrated by hindbrain systems to control ingestive behavior in response to plasma volume deficits in rats. A supracollicular transection was made which should not interfere with the neural signal of volume depletion to the hindbrain. Decerebrate (n=5) and control rats (n=7) were given subcutaneous injections of 30% polyethylene glycol (PEG) to induce hypovolemia or of isotonic saline as a control. Four hours after the injection, either water or 0.1 M NaCl was administered through an intraoral cannula, and intakes were measured. Decerebrate rats did not ingest significantly more water or saline after PEG treatment than after the control treatment, whereas control rats ingested both fluids in significantly larger volumes after PEG treatment. In another test using the same animals, heart rate was monitored after intravenous injections of phenylephrine (to raise blood pressure) and nitroprusside (to lower it). Similar reflexive changes in heart rate were observed in control and decerebrate rats, showing that baroreflex function was not impaired by decerebration. These results indicate that baroafferent signals are processed at multiple levels of the neuraxis, with hindbrain systems mediating autonomic cardiovascular reflexes in response to changes in blood pressure, and midbrain or forebrain systems mediating behavioral responses associated with thirst.     

Galanin and vasopressin response to hyperosmotic stimulation: in vitro study

Galanin (Gal)--a neuropeptide present in the nervous system and peripheral tissues--may be involved in the regulation of hypothalamo-neurohypophysial system function. It was shown that centrally injected galanin inhibits osmotically stimulated vasopressin (VP) secretion into the blood and reduces VP mRNA level in the supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei. The aim of the present study in vitro was to investigate the influence of Gal on vasopressin release from isolated rat hypothalamus (Hth), neurohypophysis (NH) or hypothalamo-neurohypophysial explants (Hth-NH). The effect of Gal on VP secretion was studied under conditions of direct osmotic (i.e., Na?-evoked) (series 1) as well as nonosmotic (i.e., K?-evoked) (series 2) stimulation. In series 3, vasopressin response to Gal was studied using the neural tissues obtained from animals drinking 2 percent NaCl solution for eight days (indirect osmotic stimulation). Gal in a concentration of 10?1? M and 10?? M inhibited basal VP release from Hth, NH and Hth-NH explants isolated from euhydrated rats as well as from Hth-NH complex of osmotically challenged animals. When the neural tissues obtained from previously salt-loaded rats were incubated in K?-enriched medium the inhibitory effect of Gal was completely blocked. It may be concluded that the effect of Gal is depending on the current functional status of the hypothalamo-neurohypophysial system.     

Afferent electrical stimulation of mesenteric nerves inhibits duodenal HCO3- secretion via a spinal reflex activation of the splanchnic nerves in the rat

The experiments were performed on male Sprague-Dawley rats anaesthetized with chloralose. Duodenal HCO3- secretion was measured in situ by pH-stat titration. The nerves bundles surrounding two randomly chosen mesenteric vessels were electrically stimulated (3 Hz, supramaximal intensity) in the afferent direction. This was done in order to mimic the intestino-intestinal spinal reflex activation of the splanchnic sympathetic fibres. The procedure reduced duodenal HCO3- secretion by 20% together with an increase in mean arterial pressure and heart rate. Duodenal HCO3- secretion decreased similarly in control rats and in those subjected to a cervical cord transection, whereas animals with bilaterally cut splanchnic nerves did not respond to such mesenteric nerve stimulation. Pharmacological pretreatment with guanethidine or yohimbine, but not prazosin, inhibited the reduction in duodenal HCO3- secretion. Thus the data suggest that electrical stimulation of mesenteric afferent nerves inhibits duodenal HCO3- secretion via a spinal reflex activation of splanchnic sympathetic nerve fibres to the duodenum, and that the response is mediated via alpha 2 adrenoceptors.     

Differential effects of estradiol on drinking by ovariectomized rats in response to hypertonic NaCl or isoproterenol: Implications for hyper- vs. hypo-osmotic stimuli for water intake

We examined the effects of estradiol on behavioral responses to osmotic challenges in ovariectomized (OVX) rats to test the hypothesis that estradiol enhances sensitivity to gradual changes in plasma osmolality (pOsm) in stimulating water intake. Despite comparably elevated pOsm after a slow infusion of 2 M NaCl, the latency to begin water intake was significantly less in estradiol-treated OVX rats compared to that in oil vehicle-treated rats. Other groups of OVX rats were injected with isoproterenol, which increases circulating angiotensin II. These rats then were given 0.15 M NaCl to drink instead of water, to prevent decreased pOsm associated with water ingestion. Isoproterenol stimulated 0.15 M NaCl intake by both groups; however, estradiol-treated rats consumed less 0.15 M NaCl than did oil-treated rats, findings that are similar to those reported when estradiol-treated rats consumed water. The estradiol enhancement of sensitivity to increased, but not to decreased, pOsm suggests that estradiol has directionally-specific effects on osmoregulatory drinking. Moreover, the estradiol attenuation of 0.15 M NaCl intake after isoproterenol suggests that estradiol effects on osmoregulatory drinking are independent of those on volume regulatory drinking.     

Vasopressinergic neurons of the supraoptic nucleus in perinatal rats: reaction to osmotic stimulation and its regulation

Osmotic stimulation (OS) of vasopressin (VP) neurons of the supraoptic nucleus (SON) promotes VP secretion and tyrosine hydroxylase (TH) expression in adult mammals. VP secretion is under a noradrenaline control, whereas the regulation of TH expression remains uncertain. This study was aimed to determine at what period of ontogenesis: (1) VP neurons begin to react to OS by modifying simultaneously VP and TH gene expression and synthesis, (2) the noradrenergic control of VP neurons is established. Rats on the 21st embryonic day (E), third postnatal day (P), P13 were salt loaded or salt loaded and treated with an antagonist (prazosin) or agonist (phenylephrine) of α1-adrenoreceptors. According to our immunocytochemical and in situ hybridization data, OS resulted in an increased amount of VP mRNA in each age group and of VP on E21 and P3. TH gene and synthesis was initially expressed under OS on P3. The number of TH-expressing neurons diminished by threefold in salt loaded rats from P3 to P13. OS combined with prazosin administration resulted in an increased level of VP mRNA on P3 and P13, but not on E21 suggesting the onset of the noradrenaline inhibitory control after birth. OS together with prazosin treatment stimulated TH expression on P3 and P13, whereas phenylephrine provided an opposite effect. Thus, VP neurons begin to react to OS by an increased VP synthesis at the end of fetal life and by the onset of TH expression shortly after birth; the expression of both substances appears to be under the inhibitory control of noradrenaline.     

Interactions between emotional stress due to fear and hypovolemic stimuli in the control of vasopressin secretion in rats

Interactions between emotional stress due to fear and hypovolemic stimuli on vasopressin secretion were studied in rats. Intraperitoneally injected dextran did not significantly change plasma osmolality and arterial blood pressure but increased blood hemoglobin and plasma vasopressin level. An i.v. infused physiological solution reversed these changes. Emotional stress due to fear acquired by learning suppressed plasma vasopressin level in dextran-injected rats. Emotional stress due to fear produced by low-frequency footshocks also suppressed the increased plasma vasopressin level. These results suggest that emotional stress due to fear interacts with afferent neural signals originating from cardio-vascular volume receptors in the control of vasopressin secretion.     

Barosensory cells in the nucleus tractus solitarius receive convergent input from group III muscle afferents and central command

Some neural mechanism must prevent the full expression of the baroreceptor reflex during static exercise because arterial blood pressure increases even though the baroreceptors are functioning. Two likely candidates are central command and input from the thin fiber muscle afferents evoking the exercise pressor reflex. Recently, activation of the mesencephalic locomotor region, an anatomical locus for central command, was found to inhibit the discharge of nucleus tractus solitarius cells that were stimulated by arterial baroreceptors in decerebrated cats. In contrast, the effect of thin fiber muscle afferent input on the discharge of nucleus tractus solitarius cells stimulated by baroreceptors is not known. Consequently in decerebrated unanesthetized cats, we examined the responses of barosensory nucleus tractus solitarius cells to stimulation of thin fiber muscle afferents and to stimulation of the mesencephalic locomotor region, a maneuver which evoked fictive locomotion. We found that electrical stimulation of either the mesencephalic locomotor region or the gastrocnemius nerve at current intensities that recruited group III afferents inhibited the discharge of nucleus tractus solitarius cells receiving baroreceptor input. We also found that the inhibitory effects of both gastrocnemius nerve stimulation and mesencephalic locomotor region stimulation converged onto the same barosensory nucleus tractus solitarius cells. We conclude that the nucleus tractus solitarius is probably the site whereby input from both central command and thin fiber muscle afferents function to reset the baroreceptor reflex during exercise.     

Bimodal effects of noxious stimuli on vasopressin secretion in rats

Effects of electric foot shocks (FSs) on plasma immunoreactive vasopressin (ir-VP) were studied in relation to FS frequency in conscious and anesthetized rats under a hypertonic condition. The plasma ir-VP level was significantly lower after low-frequency FSs (0.05-0.2 Hz) but significantly higher after high-frequency FSs (5 Hz) in conscious rats. Plasma ir-VP was significantly higher in anesthetized rats after FSs at frequencies of either 0.2 or 50 Hz compared to unshocked control rats. These data support the hypothesis that FSs activate two distinct neural mechanisms: one potentiates and the other suppresses VP secretion.     

Afferent electrical stimulation of mesenteric nerves inhibits duodenal HCO3 secretion via a spinal reflex activation of the splanchnic nerves in the rat

The experiments were performed on male Sprague-Dawley rats anaesthetized with chloralose. Duodenal HGO^- ₃ secretion was measured in situ by pH-stat titration. The nerves bundles surrounding two randomly chosen mesenteric vessels were electrically stimulated (3 Hz, supramaximal intensity) in the afferent direction. This was done in order to mimic the intestino-intestinal spinal reflex activation of the splanchnic sympathetic fibres. The procedure reduced duodenal HCO^- ₃ secretion by 20% together with an increase in mean arterial pressure and heart rate. Duodenal HGO^- ₃ secretion decreased similarly in control rats and in those subjected to a cervical cord transection, whereas animals with bilaterally cut splanchnic nerves did not respond to such mesenteric nerve stimulation. Pharmacological pretreatment with guanethidine or yohimbine, but not prazosin, inhibited the reduction in duodenal HCO^- ₃ secretion. Thus the data suggest that electrical stimulation of mesenteric afferent nerves inhibits duodenal HCO^- ₃ secretion via a spinal reflex activation of splanchnic sympathetic nerve fibres to the duodenum, and that the response is mediated via alpha₂ adrenoceptors.     

The effects of somatostatin and metiamide on tachyphylaxis of pentagastrin stimulated gastric acid and pepsin secretion in the conscious cat.

1. Pentagastrin stimulated gastric acid and pepsin secretions show parallel rates of tachyphylaxis in the conscious cat. The responses to histamine show only slight tachyphylaxis. 2. Somatostatin 10 microng.kg(-1).hr(-1) inhibits pentagastrin but not histamine stimulated acid secretion and inhibits pentagastrin stimulated pepsin secretion. 3. The inhibition of pentagastrin stimulated acid and pepsin secretion by Somatostatin delays the tachyphylaxis of these responses, but the rates of tachyphylaxis when they do subsequently occur are identical. 4. Metiamide 10 mg-kg(-1)-hr(-1) equally inhibits histamine and pentagastrin stimulated acid secretion but does not inhibit pentagastrin stimulated pepsin secretion. 5. Inhibiton of acid secretion during metiamide infusion neither prevents nor delays acid nor pepsin tachyphylaxis. 6. It is suggested that tachyphylaxis of acid and pepsin secretion is a gastrin receptor phenomenon and that Somatostatin occupies or modifies the behaviour of these receptors, preventing tachyphylaxis. Metiamide, however, exerts its action only on the histmine H2-receptor and not the gastrin receptor mechanism, and this apparently does not prevent or delay acid tachyphylaxis.     

Systolic inhibition of nociceptive responding is moderated by arousal

Inhibition of the nociceptive flexion reflex during systole could be due to activation of the arterial baroreceptors. Physiological arousal, characterized by raised blood pressure, increases afferent activity from the arterial baroreceptors but attenuates the baroreflex. This study examined the effects of arousal on systolic inhibition of the nociceptive flexion reflex in 38 adults. The threshold current to elicit the reflex in the leg was determined, and participants were stimulated at threshold intensity for 12 trials in two conditions: rest (low arousal) and mental arithmetic (high arousal). In each trial, stimulation was delivered 0 ms, 300 ms, or 600 ms after the R-wave of the electrocardiogram. Nociceptive responding was inhibited for stimulation at 300 ms after the R-wave during rest but not mental arithmetic. This moderation of systolic inhibition of nociception could be due to attenuation of the baroreflex with increased arousal.     

Physiological and pathophysiological influences on thirst

Thirst motivates animals to seek fluid and drink it. It is regulated by the central nervous system and arises from neural and chemical signals from the periphery interacting in the brain to stimulate a drive to drink. Our research has focussed on the lamina terminalis and the manner in which osmotic and hormonal stimuli from the circulation are detected by neurons in this region and how that information is integrated with other neural signals to generate thirst. Our studies of osmoregulatory drinking in the sheep and rat have produced evidence that osmoreceptors for thirst exist in the dorsal cap of the organum vasculosum of the lamina terminalis (OVLT) and in the periphery of the subfornical organ, and possibly also in the median preoptic nucleus. In the rat, the hormones angiotensin II and relaxin act on neurons in the periphery of the subfornical organ to stimulate drinking. Studies of human thirst using functional magnetic resonance imaging (fMRI) techniques show that systemic hypertonicity activates the lamina terminalis and the anterior cingulate cortex, but the neural circuitry that connects sensors in the lamina terminalis to cortical regions subserving thirst remains to be determined. Regarding pathophysiological influences on thirst mechanisms, both excessive (polydipsia) and inadequate (hypodisia) water intake may have dire consequences. One of the most common primary polydipsias is that observed in some cases of schizophrenia. The neural mechanisms causing the excessive water intake in this disorder are unknown, so too are the factors that result in impaired thirst and inadequate fluid intake in some elderly humans.     

The human nociceptive flexion reflex threshold is higher during systole than diastole

A baroreflex mechanism may explain hypertensive hypoalgesia. At rest, arterial baroreceptors are stimulated during the systolic upstroke of the pressure pulse wave. This study examined the effects of naturally occurring variations in baroreceptor activity during the cardiac cycle on an objective measure of pain, the nociceptive flexion reflex (NFR). Two interleaved up-down staircase procedures determined separate NFR thresholds during systole and diastole in 36 healthy, normotensive young adults. On odd-numbered trials, the sural nerve was stimulated electrocutaneously at R + 300 ms whereas on even-numbered trials, stimulation was delivered at R + 600 ms. The NFR threshold was higher at R + 300 ms than R + 600 ms. In contrast, stimulus intensity ratings did not differ between R + 300 ms and R + 600 ms. Stimulation of baroreceptors by natural increases in blood pressure during the systolic phase of the cardiac cycle was associated with dampened nociception.     

Effects of arterial and cardiopulmonary baroreceptor activation on simple and choice reaction times

Variations in simple reaction time over the cardiac cycle could be due to cortical inhibition associated with activation of the arterial baroreceptors. It has been proposed that higher order cognitive processing may also be modulated and, moreover, that cardiopulmonary baroreceptors may have similar inhibitory effects. This study examined arterial and cardiopulmonary baroreceptor effects on simple and choice reaction times by presenting visual stimuli at one of six intervals after the R-wave of the electrocardiogram (0, 150, 300, 450, 600, 750 ms) while participants lay supine with their legs raised or lowered. Reaction times were slower early in the cardiac cycle compared to later whereas reaction time slopes were not different. No cardiopulmonary baroreceptor effects were found. Cardiac cycle effects on reaction time are consistent with the arterial baroreceptor hypothesis and appear to be confined to lower order sensory-motor processing.     

Cardiopulmonary baroreceptors affect reflexive startle eye blink

Baroafferent signals originating from the ‘high pressure’ arterial vascular system are known to impact reflexive startle eye blink responding. However, it is not known whether baroafferent feedback of the ‘low pressure’ cardiopulmonary system loading status exerts a similar effect.Lower Body Negative Pressure (LBNP) at gradients of 0, − 10, − 20, and − 30 mm Hg was applied to unload cardiopulmonary baroreceptors. Acoustic startle noise bursts were delivered 230 and 530 ms after spontaneous R-waves, when arterial baroreceptors are either loaded or unloaded. Eye blink responses were measured by EMG, and psychomotor reaction time by button pushes to startle stimuli. The new finding of this study was that unloading of cardiopulmonary baroreceptors increases startle eye blink responsiveness. Furthermore, we replicated the effect of relative loading/unloading of arterial baroreceptors on startle eye blink responsiveness. Effects of either arterial or cardiopulmonary baroreceptor manipulations were not present for psychomotor reaction times. These results demonstrate that the loading status of cardiopulmonary baroreceptors has an impact on brainstem-based CNS processes.     

Vasopressin secretion: suppression after light and tone stimuli previously paired with footshocks in rats

Summary In an attempt to find whether vasopressin (VP) secretion is suppressed by learned emotional stress, we have given rats under a hypertonic condition simultaneously applied light and tone that had been paired previously with footshocks and have quantified immunoreactive VP (ir-VP) in the plasma. In a training session light (60 watt) and tone (2 kz) of 3-s duration which were paired with electric footshocks (50 Hz, 1-s duration) were given to rats 11 times at an interval of 30 s. Various lengths of time after the training, the rats were tested with light and tone, which were unpaired with footshocks and repeatedly applied every 15 s for 3 min in the box used for training. Hypertonic NaCl (0.5 M, 2 ml/ 100 g b. w.) was injected s. c. 30 min before testing to increase the basal level of plasma VP. After testing, plasma ir-VP was significantly less in the experimental group than in the 0-mA control group of rats that were trained without FS. The values for the experimental group were also significantly less than those of untested control rats that had been trained with FS but were not tested. Plasma osmolality and blood haemoglobin concentration were not significantly different between control and experimental groups. Plasma immunoreactive adrenocorticotrophic hormone (ir-ACTH) level was higher and motor activity as expressed by cumulative time period of body movement during testing was lower in the experimental group than in either of the control groups. The difference in plasma ir-VP between experimental and control groups was statistically significant two days but not seven days after training, whereas ACTH and motor activity in experimental groups were still significantly different from those in control groups seven days after training. The suppressive VP and augmentative ACTH responses to testing disappeared in the rats that had received light and tone repeatedly during the intervening period between training and testing. These data support the hypothesis that emotional stimuli suppress VP secretion.Supported by grants from the Ministry of Education, Science and Culture, Japan