Development of hypertension in rats during chronic exposure to cold.

Resting systolic, diastolic, and mean blood pressures (MBP), as well as heart rates, of unanesthetized, unrestrained, cold-acclimated (CA, 4 wk, 6 degrees C) rats were measured by direct arterial cannula and compared with those of controls maintained at 25 degrees C. Exposure to cold increased all these measurements significantly. Mean heart weight of CA rats was also increased significantly above that of controls. The responsiveness of MBP and heart rate to administration of the beta-adrenergic agonist, isoproterenol (3, 5, and 8 micrograms/kg ip), to unanesthetized, unrestrained, CA rats during exposure to air at 6 degrees C was similar to, and possibly less than, that of warm-acclimated (WA) rats measured at 25 degrees C. Acute administration of the alpha-adrenergic agonist, phenylephrine (100 micrograms/kg ip), to CA rats while in air at 6 degrees C induced less of a change in MBP from pretreatment level than was observed in WA rats. However, no differences were observed between groups when changes in heart rate from pretreatment level were compared. A similar statement may be made for a higher dose of phenylephrine (150 micrograms/kg ip), although MBP were elevated to higher levels in both groups with the higher dose. Abrupt exposure of WA rats to cold (6 degrees C) resulted in a sharp increase in heart rate and a more gradual increase in MBP over a period of 1 h. Removal of CA rats from 6 to 25 degrees C resulted in a gradual decrease in heart rate with no significant change in MBP during the ensuing hour.(ABSTRACT TRUNCATED AT 250 WORDS)

Cephalic reflexes: their role in digestion and possible roles in absorption and metabolism.

Stimulation of the oral cavity immediately elicits salivation, gastric acid secretion and pancreatic exocrine and endocrine secretions that serve to prepare the alimentary canal for digestion, transport and utilization of ingested nutrients. Oropharyngeal-stimulated responses are reliably initiated by the taste and smell of food. These gastrointestinal reflexes, often referred to as anticipatory or cephalic phase responses, are mediated by the autonomic nervous system and are believed to be independent of the postabsorptive effects of ingested nutrients. A common pathway used by cephalic phase responses to trigger gastrointestinal secretions is the vagus. Several studies have also demonstrated that cephalic stimulation activates both the sympathetic and parasympathetic nervous systems and thus, many cephalic-metabolic reflexes may arise indirectly from more general physiological changes that accompany oropharyngeal stimulation. The present studies suggest that oral stimulation results in alterations in intestinal function. Specifically, oropharyngeal stimulation of conscious, unrestrained rats with sucrose increases the uptake of radioactive glucose from the small intestine into the hepatic portal blood.

Effects of dietary salt, fat, and carbohydrate on fluid and salt consumption of rats.

Studies were conducted to investigate the influence of dietary sodium chloride (NaCl), fat, and carbohydrate on NaCl preferences of rats. Both long-term (48 hr) and short-term (20 min) two-bottle preference tests were used. Rats were fed either a basal salt (1.17%) diet or high salt (8.37 to 8.59%) diets in which the level of sucrose was modified by substitution with either corn oil or cornstarch. Water and NaCl consumption was measured in 48-hr two-bottle preference tests using various concentrations (0.005 to 0.5 M) of NaCl paired with water. The results of these studies suggest that changes in fluid consumption were accounted for by an increase in water consumption which was inversely related to the sucrose to starch ratio and directly related to the NaCl content of the diets. A reduction in NaCl consumption was observed only during the short-term (20 min) tests.

Response to thermal stress in the rat following acute administration of ethanol.

Several thermoregulatory responses (i.e., colonic temperature, tail-skin temperature, rate of oxygen consumption) were measured in the rat following acute administration of either saline or 3 g ethanol/kg BW i.p. at ambient temperatures (Ta) of 17, 25, and 32 degrees C. The magnitude of the ethanol-induced, hypothermic response was inversely related to Ta, with the decrease in colonic temperature (Tco) at 120 min postinjection ranging from 0.1 degree C at Ta 32 degrees C to 3.2 degrees C at Ta 17 degrees C. Depression of rate of oxygen consumption (heat production) was a major factor contributing to ethanol-induced hypothermia, with no observed differences in heat loss as assessed by differences in the responses of tail skin temperature. Thermoregulatory responses were also measured following acute administration of ethanol at Ta 25 degrees C and immediate exposure to either Ta 32 or 17 degrees C. The increase in Tco of ethanol-treated rats was delayed compared to controls following exposure to 32 degrees C. In addition, tail skin temperature and rate of oxygen consumption of treated rats were significantly lower. The delayed rise in Tco is most likely the result of a reduction in rate of oxygen consumption. When administered ethanol at Ta 25 degrees C and then exposed to Ta 17 degrees C, the rats exhibited a significant recovery from the metabolic depression that characterized the administration of ethanol during exposure to Ta 17 degrees C. This was most likely related to differences in the clearance of ethanol from blood. The results of this study are consistent with the suggestion that the physiologically significant inhibitory effect of acute administration of 3 g ethanol/kg BW on thermoregulatory responses of rats is manifested at the level of heat production rather than heat loss and that the effect is exacerbated by a reduction in Ta.

On the mechanism of serotonin-induced dipsogenesis in the rat.

Subcutaneous administration of l-5-hydroxytryptophan (5-HTP), the precursor of serotonin, to female rats induces copious drinking accompanied by activation of the renin-angiotensin system. Neither a reduction in blood pressure nor body temperature accompanied administration of 5-HTP. The objective of the present study was to determine whether serotonin-induced dipsogenesis, like that of 5-HTP, is mediated via the renin-angiotensin system. Serotonin (2 mg/kg, SC)-induced drinking was inhibited by the dopaminergic antagonist, haloperidol (150 micrograms/kg, IP), which also inhibits angiotensin II-induced drinking. Both captopril (35 mg/kg, IP), an angiotensin converting enzyme inhibitor, and propranolol (6 mg/kg, IP), a beta-adrenergic antagonist, blocked serotonin-induced dipsogenesis. The alpha 2-adrenergic agonist, clonidine (6.25 micrograms/kg, SC), which suppresses renin release from the kidney, attenuated serotonin-induced water intake. The dipsogenic responses to submaximal concentrations of both serotonin (1 mg/kg, SC) and isoproterenol (8 micrograms/kg, SC) were additive rather than interactive suggesting that similar pathways mediate both responses. The serotonergic receptor antagonist, methysergide (3 mg/kg, IP), inhibited serotonin-induced drinking but had no effect on isoproterenol (25 micrograms/kg, SC)-induced dipsogenesis. However, neither serotonin (2 mg/kg, SC) nor isoproterenol (25 micrograms/kg, SC)-induced drinking was inhibited by cinanserin (25 micrograms/kg, IP). These data indicate that serotonin induces drinking in rats via the renin-angiotensin system. However, the results of the studies using methysergide suggest that serotonin appears to act at a point prior to activation of beta-adrenoceptors in the pathway leading to release of renin from the kidneys.

Effect of acute administration of isoproterenol and angiotensin II, separately and in combination, on water intake and blood pressure of rats.

The effects of administration of isoproterenol, a beta-adrenergic agonist, and angiotensin II, a peptide, separately and in combination, on water intake and blood pressure of rats were studied. The results of 6 factorially designed studies in which 4 different doses of each compound were administered revealed that water intakes increased directly with the logarithm of increasing doses of each. The effect of simultaneous administration of the 2 compounds on water intake was additive at submaximal doses of each. No interactive effects on water intake were observed when the 2 compounds were administered simultaneously in any study. Reduction in urine output appears to be a more sensitive response to administration of isoproterenol than increase in water intake since it was virtually abolished at a dose (2.5 micrograms/kg SC) that had no effect on water intake. The lowest doses of angiotensin II (25 and 50 micrograms/kg SC) had no significant effect on either water intake or urine output. The effect of simultaneous administration of both compounds on urine output was essentially the same as that accompanying administration of isoproterenol alone. Following administration of angiotensin II (150 micrograms/kg, SC) mean systemic blood pressure of unanesthetized, chronically cannulated rats reached maximal levels within 5 min and returned to pretreatment control level by 60 min. Following administration of isoproterenol (25 micrograms/kg, SC), mean systemic blood pressure decreased within 5 min, was maximally depressed by 30 min and had returned halfway to the pretreatment control level by 60 min. Simultaneous administration of isoproterenol and angiotensin II failed to induce a significant change in blood pressure. These results are of particular interest since they show that neither the pressor effect of angiotensin II nor the depressor effect of isoproterenol is essential for the induction of drinking by these 2 compounds.

Dependence of beta-adrenergic responsiveness on thyroid state of male rats.

The dose of thyroxine (0-150 micrograms/kg body weight) administered subcutaneously daily to surgically thyroidectomized male rats was correlated significantly with their metabolic activity as assessed by rate of oxygen consumption and colonic temperature. There was a significant dose-dependent increase in total serum thyroxine, triiodothyronine and reverse-triiodothyronine concentrations with increasing doses of thyroxine administered. The cardiostimulatory response to administration of isoprenaline (8 micrograms/kg body weight s.c.) was also correlated directly with the dose of thyroxine administered. The concentration of cardiac beta-adrenoceptors in these animals was correlated significantly with total serum thyroxine and triiodothyronine concentrations, basal heart rate, and the chronotropic response to administration of isoprenaline.

Water deprivation-induced drinking in rats: role of angiotensin II.

The role of angiotensin II in the control of water intake following deprivation of water for varying lengths of time was studied. Male rats were deprived of water for 0, 12, 24, 36, or 48 h. Water intakes were measured with and without pretreatment with the angiotensin I-converting enzyme inhibitor, captopril (50 mg/kg, ip). Captopril had no significant effect on water intake following either 0 or 12 h of water deprivation. However, captopril significantly attenuated water intake following 24-48 h of water deprivation with the magnitude of the attenuation increasing as the length of the period of water deprivation increased. Plasma renin activity was significantly increased over control levels after 24-48 h of water deprivation but not after 12 h of water deprivation. Plasma renin activity tended to increase as the length of the water-deprivation period increased. Serum osmolality and sodium concentration were significantly increased over control levels following 12-48 h of water deprivation. Serum osmolality and sodium concentration failed to show any further increases with increasing length of water deprivation beyond the increases following 12 h of water deprivation. The data indicate that the water intake of water-deprived rats can be divided into an angiotensin II-dependent component and angiotensin II-independent component. The angiotensin II-independent component appears to be more important in the early stages of water deprivation whereas the angiotensin II-dependent component becomes more important as the length of the water-deprivation period increases.

Renal-thyroid interrelationship in normotensive and hypertensive rats.

An increase in the weight of the thyroid gland, accompanied by reductions in uptake and release of 1311, serum T3 and reverse T3 concentrations and an increase in serum TSH concentration, occurs during the development of renal hypertension in rats. Renal function, as assessed by BUN, serum and urinary creatinine concentrations, and ability to concentrate urine during dehydration, is depressed. The kidneys of both normotensive and renal hypertensive rats contain a thyroid depressing factor (TDF) which inhibits uptake of 131I both in vivo and in vitro as well as inhibiting the lactoperoxidase enzyme in vitro. The kidneys of renal hypertensive rats contain less, and the serum more, TDF than those of normotensive rats. Whether TDF is produced as a result of hypertension per se or as a result of renal damage accompanying hypertension remains for further study. TDF appears to be a peptide with a molecular weight in the range of 400 to 419 daltons.

Interrelationships among blood pressure, renal function, thyroid activity and renal thyroid depressing factor in renal hypertensive rats.

Renal hypertension in rats was accompanied by both a decrease in thyroid activity (i.e., decreased total serum triiodothyronine concentration, increased serum thyroid-stimulating hormone concentration and increased thyroid weight) and renal function (i.e., reduced renal concentrating ability, increased blood urea nitrogen and serum creatinine concentrations). Systolic blood pressure, thyroid activity and level of renal function of individual hypertensive rats correlated significantly with the content of a thyroid-depressing factor in their kidneys. The results suggest, but do not prove, that a factor released by the kidneys during elevation of blood pressure and/or reduction in renal function may reduce thyroid function in the rat.

Peripheral conversion of L-5-hydroxytryptophan to serotonin induces drinking in rats.

Female rats administered serotonin (0.25 to 4.0 mg/kg, s.c.) showed a dose-dependent increase in water intake. The dipsogenic response was nearly maximal when 2 mg/lg was administered s.c. and plateaued by 2 hr after treatment. l-5-Hydroxytryptophan (5-HTP), the precursor of serotonin, is also a potent dipsogen which induces drinking by way of the renin-angiotensin system. The possibility that the dipsogenic activity of 5-HTP is dependent on decarboxylation to serotonin was the objective of these studies. Either benserazide (30 mg/kg. s.c.), a central and peripheral decarboxylase inhibitor, or carbidopa (6.5 mg/kg, s.c.), a peripheral decarboxylase inhibitor, was administered 15 min prior to the dipsogen. Both decarboxylase inhibitors attenuated the dipsogenic response to 5-HTP (25 mg/kg, s.c.) but not to serotonin (2 mg/kg, s.c.). The peripheral serotonergic receptor antagonist, methysergide (3 mg/kg, i.p.), blocked the dipsogenic responses to both 5-HTP (25 mg/kg, s.c.) and serotonin (2 mg/kg, s.c.). There was no interaction between 5-HTP (18 mg/kg, s.c.) and serotonin (1 mg/kg, s.c.) when administered simultaneously with respect to their dipsogenic effects. Thus, the drinking response accompanying administration of 5-HTP occurs following peripheral conversion to serotonin which, in turn, activates peripheral serotonergic receptors. The mechanisms(s) by which activation of peripheral serotonergic receptors increases water intake is not known, but appears to involve release of renin from the kidney.

Effects of serotonin and L-5-hydroxytryptophan on plasma renin activity in rats.

The effects of dipsogenic doses of l-5-hydroxytryptophan (5-HTP) and serotonin on plasma renin activity (PRA), blood pressure, and body temperature were determined in unanesthetized female rats. Both serotonin (2 mg/kg, s.c.) and 5-HTP (25 mg/kg, s.c.) induced six-fold increases in PRA measured 1 hr after drug administration. The central and peripheral decarboxylase inhibitor, benserazide (30 mg/kg, s.c.), as well as the peripheral decarboxylase inhibitor, carbidopa (6.5 mg/kg s.c.), prevented the increase in PRA associated with administration of 5-HTP. This suggests that 5-HTP must be converted to serotonin peripherally to increase PRA. At the doses used, serotonin decreased mean blood pressure and colonic temperature of unanesthetized rats while 5-HTP was without effect. The increase in PRA induced by 5-HTP does not appear, therefore, to be a response to either hypotension or a decrease in colonic temperature. Since 5-HTP must be converted to serotonin to initiate both a drinking response and an increase in PRA, the results suggest that the decrease in blood pressure and colonic temperature following administration of serotonin may not be important in induction of the drinking response and the increase in PRA. The mechanism by which activation of the renin-angiotensin system occurs following peripheral administration of either 5-HTP or serotonin remains for further study.

L-5-Hydroxytryptophan-induced drinking in rats: possible mechanisms for induction.

Administration of L-5-hydroxytryptophan (25 mg/kg body weight, SC) to female rats resulted in copious drinking. The dipsogenic response to administration of L-5-hydroxytryptophan (5-HTP) was blocked by propranolol (6 mg/kg body weight, IP), a beta-adrenergic antagonist, and captopril (35 mg/kg body weight, IP), an angiotensin converting enzyme inhibitor. In addition, clonidine (12.5 and 25 microgram/kg body weight, IP), a central alpha-adrenergic agonist known to inhibit renin release, attenuated drinking during 1, 2 and 3 hours after 5-HTP was administered. These results suggest that 5-HTP-induced drinking is mediated by way of the renin-angiotensin system. Haloperidol (150 microgram/kg body weight, IP), a dopaminergic antagonist, also attenuated the dipsogenic response to administration of 5-HTP. In addition, incremental reductions in 5-HTP-induced drinking with increasing doses of spiperone (37.5 to 150 microgram/kg body weight, IP), a more potent dopaminergic antagonist, were demonstrated. Thus, the dipsogenic response to administration of 5-HTP to rats is dependent on both the renin-angiotensin system and an intact dopaminergic pathway.

Use of lactoperoxidase-catalyzed iodination of L-tyrosine to assess antithyroid activity of a factor present in certain tissues of mammals.

A factor, present in the kidneys of rats, has been shown to inhibit thyroid activity both in vivo (rat) and in vitro (porcine and bovine thyroid slices). The aim of the present study was to quantitate the inhibitory activity of this factor in renal and other tissues from rats by means of the lactoperoxidase-catalyzed iodination of L-tyrosine. Supernatants of renal homogenates (10,000, 27,000 and 35,000 g) obtained from rats inhibited this in vitro iodination system as well as the lactoperoxidase-catalyzed oxidation of guaiacol. The 35,000-g supernatant from both renal and hepatic tissues contained the greatest inhibitory activity per milligram of protein. Supernatants of homogenates (35,000-g fraction) from testes contained one eighth to one third the activities of liver and kidney, respectively. Kidneys of other species including fetal goat, mature goat, fetal lamb, mature monkey and mature dog, were also shown to contain the thyroid-depressing factor. These studies suggest that the thyroid-depressing factor is found mainly in liver and kidneys of the rat and that it is not unique to the rat but is present in the kidneys of other species as well.

Normal excretion of m-hydroxymandelic acid in hypertensive patients.

o-Hydroxymandelic acid (OHMA), m-hydroxymandelic acid (MHMA) and p-hydroxymandelic acid (PHMA) were measured in the urine of 42 normotensive and 54 hypertensive patients. Patients having high urinary MHMA levels were all found to be ingesting medications containing m-synephrine (phenylephrine). These patients also had high levels of urinary m-synephrine which was excreted as the glucuronide. When patients ingesting m-synephrine were excluded from the analysis, no significant differences were observed between the two groups for the urinary excretion of OHMA, MHMA and PHMA.

Dipsogenic effect of L-5-hydroxytryptophan in rats.

The precursor of serotonin, l-hydroxytryptophan (5HTP), is a potent dipsogen in the rat. Peripheral administration of increasing doses of this compound increased water intake in a dose-dependent fashion. Peripheral administration of other analogs of tryptophan, including d-tryptophan, l-tryptophan and acetyltryptophan, failed to affect water intake at a dose at which 5HTP induced maximal drinking (25 mg/kg, SC). The dipsogenic effect of melatonin, one of the metabolites of serotonin, was also tested. At any of 6 different doses (0.5 to 50 mg/kg, SC), melatonin failed to affect water intake in the rat. The mechanism by which 5HTP induces drinking is not known with certainty but could involve its conversion to serotonin, a known dipsogenic agent.

Use of an iodide-specific electrode to study lactoperoxidase-catalyzed iodination of l-tyrosine.

An in vitro method employing an iodide-specific electrode for monitoring lactoperoxidase-catalyzed iodination is described. The method utilized lactoperoxidase, potassium iodide, and a glucose--glucose oxidase system for the generation of hydrogen peroxide and l-tyrosine. As iodination of l-tyrosine proceeded, the free iodide concentration in solution decreased and was monitored by an iodide-specific electrode. The iodide electrode was reliable when compared to a 131I-method for measuring free iodide changes in solution. Increasing concentrations of resorcinol, a well-known inhibitor of thyroid peroxidase-catalyzed iodination, in the reaction mixture resulted in graded inhibition of the initial rate of lactoperoxidase-catalyzed l-tyrosine iodination. This in vitro system can be used to assess inhibitory activity of various antithyroid substances.