Blood pressure and endocrine responses of healthy subjects in cold pressor test after acutely increased dietary sodium intake.

The objective of the study was to compare blood pressure and endocrine responses in a cold pressure test in young healthy subjects who had shown increased blood pressure during an acutely increased sodium intake. Subjects (n = 53) added 121 mmol sodium into their normal diet for one week. If the mean arterial pressure had increased by a minimum of 5 mmHg compared to the control measure, they were selected for the experiments. The selected subjects (n = 8) were given 121 mmol supplemental sodium d-1 for 14 days after which they immersed the right hand into a cold (+10 degrees C) water bath for 5 min. The blood pressure increased (P < 0.05) during the test and was independent of the sodium intake. The plasma noradrenaline increased from 2.41 +/- 0.38 nmol l-1 to 2.82 +/- 0.42 nmol l-1 (P < 0.05) with normal diet and from 1.85 +/- 0.29 nmol l-1 to 2.40 +/- 0.37 nmol l-1 (P < 0.05) with high sodium diet. The starting concentrations and the endpoint concentrations were statistically similar. The plasma levels of natriuretic peptides (NT-proANP, ANP and BNP) did not change during the test, and the concentrations were independent of the sodium diet. To conclude, acutely increased sodium intake does not change blood pressure or hormonal responses in a cold pressor test in young healthy subjects.

Are the blood pressure and endocrine responses of healthy subjects exposed to cold stress altered by an acutely increased sodium intake?

In the study reported here, we examined blood pressure and endocrine responses in cold conditions during salt load in young healthy subjects who had previously shown increased resting blood pressure during acutely increased sodium intake. Subjects (n = 53) added 121 mmol sodium into their normal diet for 1 week. If their mean arterial pressure had increased by a minimum of 5 mmHg compared to the previous measure they were selected for subsequent experiments. The subjects (n = 8) were given 121 mmol supplemental sodium.day-1 for 14 days. They were then put into a wind tunnel for 15 min (temperature--15 degrees C, wind speed 3.5.ms-1). Their blood pressure increased (P < 0.05) during the cold exposure, independent of the sodium intake. Their mean (SEM) plasma noradrenaline increased from 3.58 (0.62) nmol.l-1 to 5.61 (0.79) nmol.l-1 (P < 0.05) when the subjects were given a normal diet, and from 2.45 (0.57) nmol.l-1 to 5.06 (0.56) nmol.l-1 (P < 0.05) when the subjects were given an elevated sodium diet. The starting concentrations and the endpoint concentrations were statistically similar. The plasma levels of the N-terminal fragment of pro-atrial natriuretic peptide decreased during the whole-body cold exposure: with the sodium load the change was from 256.6 (25.5) nmol.l-1 to 208.0 (25.3) nmol.l-1, and with the normal diet, from 205.8 (16.4) nmol.l-1 to 175.1 (16.1) nmol.l-1. The haematocrit and red blood cell count increased (P < 0.05) with normal and elevated sodium diet in cold conditions, but haemoglobin increased (P < 0.05) only with high salt in cold conditions. To conclude, acutely increased sodium intake does not change the blood pressure response or hormonal responses to exposure to acute cold stress in healthy subjects.

Localization of salmon cardiac peptide (sCP) in the heart of salmon (Salmo salar L.).

We have previously cloned and characterized a novel cardiac hormone from the salmon (Salmo salar) which has a uniquely heart-specific distribution and a low structural similarity with any other known natriuretic peptides. Specific antibodies were raised in goat against the salmon cardiac peptide. For localization and quantification, four different methods were applied: immunohistochemistry (avidin-biotin peroxidase), transmission electron microscopy, cryoimmunoelectron microscopy (protein A-gold), and a specific radioimmunoassay. Both atrial and ventricular myocytes stained immunohistochemically. The staining was similar in all myocytes and no specific myoendocrine cells were found. Within a single myocyte, both atrial and ventricular, the staining was stronger near the nucleus. Transmission electron microscopy revealed that both the atrium and the ventricle contained small sarcoplasmic granules of similar type with a diameter of 100 to 200 nm and an electron-dense core with a clear halo. The granules were typical vesicles which can be found in secretory cells utilizing the regulatory pathway. The highest number of granules was found near the nucleus, but granules were located also near the Golgi apparatus, between myofilament bundles, and in subsarcolemmal positions. Gold particles, conjugated to antibodies raised against the salmon cardiac peptide, were deposited on similar sarcoplasmic granules found in transmission electron microscopy. Among the sarcoplasmic granules with gold particles there were granules which did not show any cardiac peptide immunoreactivity. A significantly (Student's t test, P < 0.05) higher concentration of cardiac peptide was found in the heart atrium than in the ventricle, 16.2 +/- 3.5 pmol/mg tissue (n = 8) and 4.5 +/- 1.7 pmol/mg tissue (n = 8), respectively. The findings show that the salmon cardiac peptide is localized in secretory granules in both compartments of the heart. The morphology of the granules suggests that both the atrium and the ventricle utilize the regulatory pathway to release salmon cardiac peptide.

Novel salmon cardiac peptide hormone is released from the ventricle by regulated secretory pathway.

We used the secretion of the novel salmon cardiac peptide (sCP) as a model to examine the mechanisms of ventricular hormone release. Mechanical load increased dose dependently the secretion of immunoreactive sCP from isolated perfused salmon ventricle, with 3. 3-fold increase when a load of 13 cmH(2)O was applied. Endothelin-1 (5 nmol/l) was also able to rapidly increase the secretion of sCP. The released peptide corresponded to the biologically active sCP-29, whereas the large ventricular storage consisted of pro-sCP-sized material. With the use of immunoelectron microscopy, a large number of granules containing immunoreactive sCP could be detected in salmon ventricle. As judged by RNA blot analysis, there was very active basal expression of the sCP gene in the ventricle, which was not increased by mechanical load of up to 2-h duration. Our results show that the ventricle actively expresses the gene of sCP, stores the prohormone in secretory granules, and releases the peptide in response to mechanical load and endothelin-1. Thus the salmon ventricle uses the regulated pathway to produce and release a hormone structurally related to the mammalian natriuretic peptides.

Blood pressure and hormonal responses to short whole body cold exposure in subjects with high dietary salt intake.

The objective of the present study was to test a hypothesis that a high dietary salt intake potentiates a cold induced increase in blood pressure in normotensive men. Male subjects (n = 12) were given 7 g day-1 sodium chloride during the cold months of the year, divided in 3-4 doses per day and dissolved in water, for 14 days additional to their normal diet which contained on the average 9.7 g sodium chloride per day. The same subjects, having their normal diet, served as controls. The resting blood pressure was measured on the fourteenth day seven times at the intervals of five minutes in a climatic chamber in thermoneutral conditions. Then the subjects, wearing a three-layer winter clothing, moved into a wind tunnel (-15 degrees C, air velocity 3.5 ms-1) in which they stayed for fifteen minutes and the blood pressure was recorded at the intervals of three minutes. After the cold exposure, the subjects moved back into the climatic chamber for 30 min and the blood pressure was measured as before the cold exposure. Blood samples were drawn before and after the experiment for ion and hormone measurements. A 12 h urine sample was collected just prior to the cold exposure. A significant difference both in systolic (7 mmHg) and in diastolic (7 mmHg) blood pressure was found between a salt load group and control group under thermoneutral conditions, repeatedly measured over 30 min (paired Student's t-test; p < 0.05). During the whole body cold exposure, blood pressure significantly increased both with and without the extra salt load (repeated measures ANOVA, Student-Newman-Keuls; p < 0.05). The level to which the mean arterial pressure increased during the exposure was independent of the salt intake and the profile of the mean arterial pressure curve was similar in both groups. The systolic pressure increased by a 25 mmHg in both groups during the cold exposure. The increase in the diastolic pressure was significantly (paired Student's t-test, p < 0.05) higher in the high salt group (18 +/- 4 mmHg) than in the control group (12 +/- 3 mmHg) thus supporting partly our hypothesis. After the two-week high salt intake, serum Na+, K+, Cl-, Hct, and plasma Hb were at the similar level as before the extra salt intake. Plasma renin activity, NT-proANP, ANP, and serum aldosterone were not different between the groups, both before and after the cold exposure. The main findings are: 1) the mean arterial pressure increases to the same level and in the same manner independent of the salt load during a short whole body cold exposure and 2) in cold the diastolic blood pressure increases significantly more in people under a very high salt diet.

A novel cardiac hormone related to A-, B- and C-type natriuretic peptides.

Mechanisms acting against accumulation of volume are important in pathophysiological situations with volume and salt overload, such as congestive heart failure. Osmoregulating animals that migrate between environments with high and low salinity are ideal models for studying the defence mechanisms against volume gain. We have now cloned and sequenced from salmon (Salmo salar) a cDNA encoding a novel vasorelaxant cardiac hormone of 29 amino acids which is produced by proteolytic processing of a 148-residue preprohormone. Structural and biological results, as well as its distribution indicate that it belongs to an unrecognized family related to natriuretic peptides, perhaps representing an ancestor of ANP and BNP. We have synthesized the 29-amino acid hormone and set up a specific radioimmunoassay. The distribution of the mRNA and peptide is strictly restricted to the heart, with high levels both in the atrium and ventricle in various fish species. The hormone relaxes aortic smooth muscle derived from salmon at nanomolar concentrations. Its release from isolated perfused salmon ventricle is very sensitive to mechanical load: a 10 mmHg load induces a rapid 5-fold increase in hormone release. Our results indicate that the novel cardiac hormone has an important role in fish volume regulation. They also demonstrate that mechanical stimuli have been central to volume regulation since early evolution.

Brain natriuretic peptide (BNP) gene expression in the heart of vasopressin-deficient Brattleboro rats.

Brain natriuretic peptide (BNP) messenger RNA was measured with a semiquantitative method from heart auricles and ventricles of vasopressin-deficient Brattleboro rats (DI) and from desmopressin treated Brattleboro rats (DI + DDAVP). Desmopressin had been injected peripherally and Long-Evans rats (LE) served as controls. The 3-day substitution treatment had shifted the fluid balance of DI almost to that of LE. In the present study, the amount of BNP mRNA, normalized to the glyceraldehyde-3-phosphate dehydrogenase mRNA content, was constant in all three groups in the right auricle. No changes were when the right auricular and left auricular mRNA levels were compared within each group. In the left auricle, desmopressin treatment increased significantly (P < 0.05) the amount of BNP mRNA compared with that of LE rats (from 1.09 +/- 0.21, n = 7 to 1.72 +/- 0.17, n = 8, arbitrary units). In all groups, the left ventricle had significantly (P < 0.05) higher mRNA content than the right ventricle (LE: 2.24 +/- 0.23 vs. 0.67 +/- 0.13, n = 6; DI: 2.30 +/- 0.60 vs. 0.33 +/- 0.05, n = 8; DI + DDAVP: 2.36 +/- 0.29 vs. 0.37 +/- 0.07, n = 10). In the right ventricle, both DI and DI + DDAVP rats had significantly (P < 0.05) lower mRNA content than LE rats (0.33 +/- 0.5 vs. 0.67 +/- 0.13 and 0.37 +/- 0.07 vs. 0.67 +/- 0.13, respectively). To conclude, these findings suggest that brain natriuretic peptide gene expression dissociates from, or rapidly adapts to, the chronic effects of peripheral desmopressin treatment which have shifted the fluid balance to almost normal in Brattleboro rats. The left ventricular pressure appears to regulate the brain natriuretic peptide gene expression.

Atrial plasma ANP and NH2-terminal proANP during right atrial pressure increase in humans.

To compare plasma NT-proANP, a stable and biologically inactive N-terminal portion of ANP prohormone, with the known plasma ANP response to increased right atrial pressure a Swan-Ganz catheter was inserted into the right atrium of five normal healthy male volunteers. The elevation of right atrial pressure was produced by a head-down tilt after a hypertonic saline infusion. Blood samples were drawn from the lumen of the right atrium. After 5 min of starting the tilt the right atrial pressure had increased from 7.0 +/- 1.0 to 11.6 +/- 0.9 mmHg (P < 0.05) and then began to normalize in spite of the constant tile. Atrial plasma ANP increased in relation to the pressure increase and peaked at 15 min after the start of the tilt. The change was from 27.9 +/- 6.5 to 53.9 +/- 9.7 pmol L-1 (P < 0.05). Atrial plasma NT-proANP increased significantly from 357 +/- 91.2 to 529.1 +/- 116.0 pmol L-1 (P < 0.05) at 10 min and remained high throughout the experiment. The molar ratio of NT-proANP to ANP varied in atrial plasma from 9.5 +/- 1.2 to 13.9 +/- 2.7 showing that the plasma clearance of ANP from plasma was much higher than that of NT-proANP.

Hemodynamic control of atrial natriuretic peptide plasma levels in neonatal respiratory distress syndrome.

To evaluate the contribution of the pulmonary and ductal hemodynamics on the cardiac atrial natriuretic peptide (ANP) synthesis and release in neonatal respiratory distress syndrome, serial blood samples for plasma C-terminal end, and the more stable N-terminal end (NT-proANP) of the propeptide were obtained. Simultaneous evaluation of the systolic pulmonary artery pressure (PAP) and magnitude of ductal shunting by the Doppler method were made of 37 distressed infants during the first 4 days of life. Both plasma ANP and NT-proANP rose after birth, peaked at 48 hours of age, and correlated significantly (r = 0.66; p < 0.001; n = 78) with each other. The initially high systolic PAP and, since the systemic arterial pressure (SAP) did not change, the PAP/SAP ratio declined slowly during the study period, as did the magnitude of ductal left-to-right shunting after an initial increase during the first hours after birth. Plasma NT-proANP had a positive correlation to the magnitude of ductal left-to-right shunting both during the first 2 and 4 days of life, but did not correlate with PAP, SAP, or PAP/SAP ratio during the same time periods. Eight infants with delayed closure of the ductus maintained elevated plasma NT-proANP values after the second day of life.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrial natriuretic peptide and its mRNA in heart and brain of vasopressin-deficient Brattleboro rats.

To understand the secretion and synthesis of atrial natriuretic peptide we measured immunoreactive atrial natriuretic peptide from plasma, heart tissues and brain areas, and ANP mRNA was determined from heart auricles and ventricles of vasopressin-deficient Brattleboro rats (DI) and from desmopressin treated Brattleboro rats (DI+DDAVP). Long-Evans rats (LE) served as controls. DI+DDAVP rats were given for 3 days sc. injections of 0.5 micrograms 1-desamino-8-D-arginine vasopressin in 1 mL saline twice a day. The rats were housed in single metabolic cages and urinary output and water intake were measured daily. All the body and organ weight parameters were similar in the three groups when the rats were killed. No change was seen in the plasma ANP level between the groups. The right ventricle of DI+DDAVP rats had significantly (P < 0.05) higher concentration of ANP than LE rats (15.8 +/- 4.4 vs. 3.4 +/- 0.6 ng mg-1 tissue). The left ventricle of DI and DI+DDAVP had significantly (P < 0.05) lower amounts of ANP mRNA than LE rats (0.5 +/- 0.2 vs. 1.3 +/- 0.2 and 0.5 +/- 0.1 vs. 1.3 +/- 0.2 arbitrary units). In the hypothalamus, the ANP concentration was significantly (P < 0.05) lower both in DI and in DI+DDAVP rats than in LE rats (9.3 +/- 1.3 vs. 14.5 +/- 1.6 and 6.1 +/- 0.6 vs. 14.5 +/- 1.6 pg mg-1 tissue). To conclude, although the water intake and urinary output of DI rats were changed towards normal with desmopressin treatment, the heart ventricular and hypothalamic ANP did not parallel the change.(ABSTRACT TRUNCATED AT 250 WORDS)

Block of stretch-activated atrial natriuretic peptide secretion by gadolinium in isolated rat atrium.

1. Isolated superfused rat atrial preparations were used to study the mechanism of stretch-induced atrial natriuretic peptide (ANP) secretion. The stretch of the atrial myocytes was induced by raising the intra-atrial pressure. The secretion rates were analysed by measuring ANP concentrations from the superfusate fractions by radioimmunoassay. 2. The effect of gadolinium, a blocker of stretch-activated ion channels, on stretch-induced and basal ANP secretion was investigated by superfusing the atrial preparation with 5, 20 or 80 microM GdCl3. Gadolinium decreased stretch-induced ANP secretion in a dose-dependent manner, but did not affect basal secretion. 3. Because high concentrations of gadolinium may block voltage-gated calcium channels, we tested whether the selective blockers of L-type (diltiazem) and T-type (NiCl2) calcium channels affect the stretch-stimulated ANP release. Neither diltiazem at 3 microM nor NiCl2 at 50 microM affected stretch-induced ANP release in paced atrial preparation. 4. Gadolinium, but not diltiazem, also inhibited stretch-stimulated ANP secretion in non-paced, quiescent atria. 5. The findings that ANP release is inhibited by Gd3+, but not by diltiazem or NiCl2, and that the stretch-induced secretion in quiescent atria is also inhibited by Gd3+, suggest that stretch-activated ion channels are involved in the regulation of stretch-induced ANP release.

Effect of ryanodine on atrial natriuretic peptide secretion by contracting and quiescent rat atrium.

To elucidate the mechanism involved in the release of atrial natriuretic peptide (ANP), we studied the importance of ryanodine-sensitive Ca2+ release in stretch-secretion coupling. The experiments were made with a left atrial preparation, where the stretch of myocytes was induced by changing the intra-atrial pressure. When external pacing was not applied, the atrial preparation was not spontaneously contracting, and it was therefore possible to investigate the secretory mechanism in the quiescent atrium. The superfusate was collected in 2-min fractions and assayed for ANP immunoreactivity. Filtration analysis revealed that the major fraction in the superfusate in all experimental situations had a similar molecular weight as the ANP 1-28. Ryanodine (1.0 microM and 0.1 microM) inhibited stretch-stimulated ANP secretion dose dependently both in paced and non-paced atrium, but did not have any effect on basal secretion. The present results support the notion that intracellular Ca2+ transients from the intracellular stores are essential for stretch-stimulated ANP secretion, independently from excitation and contraction. Basal ANP secretion is not inhibited by blocking ryanodine-sensitive Ca2+ channels, either in contracting or in non-contracting atria. In addition our results confirm that the principal stimulus for ANP secretion in response to atrial distension is the stretch of myocytes. Length shortening of myocytes is not essential for ANP release.

Plasma endothelin-1 in the neonatal respiratory distress syndrome.

In order to study the contribution of the vasoconstrictory peptide endothelin-1 (ET-1) to the elevation of the pulmonary vascular resistance in the neonatal respiratory distress syndrome (RDS) seven preterm infants were studied at 2, 24 and 48 hours of age for plasma ET-1 concentrations and systolic pulmonary artery pressure (PAP), measured by Doppler sonography. Plasma ET-1 levels were high during the first day after birth, but declined soon to normal levels thereafter. During the study period, the systolic PAP also decreased, whereas the systemic pressure remained unchanged. There were no correlations between plasma ET-1 and vascular pressures, but there was a significant association between ET-1 and the requirement of supplemental oxygen and the arterial-alveolar oxygen tension ratio. Our results thus suggest that high plasma ET-1 levels in the acute phase of RDS reflect the severity of the pulmonary disease, but may not significantly contribute to the elevation of the pulmonary vascular resistance in the RDS.

Atrial natriuretic peptide in the preterm infant. Lack of correlation with natriuresis and diuresis.

We assessed the relation of atrial natriuretic peptide (ANP) to renal function on postnatal day 2 and day 5 in preterm infants. Plasma ANP concentration was measured by radioimmunoassay in two groups of preterm infants: group 1, gestational age less than 30 weeks, n = 10; and group 2, gestational age 30-34 weeks, n = 11. The identity of the immunoreactivity as ANP-28 was confirmed by HPLC. Plasma ANP was significantly higher in group 1 than in group 2 on day 2 and day 5 (p < 0.01) and ANP concentration decreased by day 5 in both groups (group 1, p < 0.01; group 2, p < 0.02). The results showed no correlation between plasma ANP concentration and urinary sodium excretion or creatinine clearance, which may be due to a blunted renal response to ANP, but other factors may be involved also. We conclude that preterm infants are able to release large amounts of ANP, but a high plasma ANP concentration does not correlate directly with renal regulation of sodium and water balance.

Plasma ANP during hypertonic NaCl infusion in man.

To determine the relationship between hyperosmolality and immunoreactive atrial natriuretic peptide of heart atrial plasma six healthy men were given 0.06 ml kg-1 min-1 855 mmol l-1 NaCl, i.v., for 2 h. The right atrial pressure and atrial plasma atrial natriuretic peptide were measured. During the infusion, right atrial pressure was kept constant by lowering the legs of the subject in a supine position downwards if any increase in the pressure was seen. There was a significant and linear increase in atrial serum osmolality, from 288 +/- 3.3 to 307 +/- 3.2 mOsm kg-1 (P less than 0.001). No statistically significant changes in right atrial pressure were seen. Regression analysis revealed that there was a statistically significant correlation between serum osmolality and plasma ANP in three subjects (responders) (r2: 0.5241, 0.8965, 0.6695). In three other subjects (nonresponders), there was no correlation between osmolality and ANP. The mean basal osmolality of responders was 280 mOsm kg-1 and the mean basal osmolality of nonresponders was 295 mOsm kg-1. In contrast, all subjects responded with an increase in plasma ANP (P less than 0.05) after RAP had been increased by tilting the legs of the subject upwards for 30 min. We conclude that the right atrial pressure regulates the release of atrial natriuretic peptide. Serum hyperosmolality may also contribute to the regulation of atrial natriuretic peptide independently of the right atrial pressure in man.

Plasma immunoreactive atrial natriuretic peptide and vasopressin after ethanol intake in man.

To study the mechanisms of alcohol-induced diuresis, the plasma concentration of immunoreactive atrial natriuretic peptide and arginine vasopressin, serum sodium and osmolality, plasma renin activity and aldosterone, urinary sodium and volume, free water clearance, blood pressure and heart rate were measured in seven healthy men after oral intake of ethanol (1.5 g kg-1 in 6 h). Serum ethanol levels increased to 27 +/- 4 mmol l-1 (mean +/- SD) in 30 min and remained detectable for 14 h. Serum osmolality rose from 280 +/- 10 to 340 +/- 4 mosm kg-1 in 2 hours (P less than 0.01) and was 300 +/- 4 at 14 h (P less than 0.01). Formation of hypotonic urine began after the alcohol intake and resulted in a net loss of 0.9 +/- 0.1 kg water in 2 h. Free water clearance increased from -3.4 +/- 1.4 to 2.8 +/- 1.5 ml min-1 in 2 h (P less than 0.01). Plasma immunoreactive arginine vasopressin decreased from 5.7 +/- 2.1 to 3.3 +/- 1.3 ng l-1 (P = 0.05) in 30 min and increased to 17 +/- 25 and 12 +/- 10 ng l-1 at 6 and 12 h, respectively (P less than 0.05 for both). Plasma immunoreactive atrial natriuretic peptide levels decreased from 17 +/- 9 to the minimum of 11 +/- 3 ng l-1 in 2 h (P less than 0.01) and returned to the initial levels in 6 h. Serum sodium, plasma renin activity and plasma aldosterone increased maximally by 4 +/- 2, 165 +/- 153 and 143 +/- 101% (P less than 0.01 each) during 1-6 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Passive heat exposure leads to delayed increase in plasma levels of atrial natriuretic peptide in humans.

The effects of passive heat exposure on atrial natriuretic peptide (ANP) were studied in six healthy men staying in a Finnish sauna at +92 degrees C for 20 min. Their rectal temperature increased by 0.4 degrees C, and evaporative water loss was 0.92 +/- 0.14 (SD) kg. Heart rate and systolic blood pressure increased significantly during the 20-min exposure. Serum osmolality and plasma arginine vasopressin levels increased during the exposure, then declined, and increased significantly again at 90-120 min. Plasma renin activity and aldosterone increased by two- to fourfold in 20 min. Plasma ANP levels rose from 13 +/- 7 to 39 +/- 15 ng/l at 60 min and to 41 +/- 13 ng/l at 120 min (P less than 0.01 for both). We conclude that transient increases in heart rate and systolic blood pressure or changes in blood volume as inferred from the weight loss do not contribute to the increased plasma ANP levels observed after the heat exposure. Instead, increased secretions of pressor hormones could explain the elevated plasma ANP levels observed after the thermal stress.

Daily patterns of plasma atrial natriuretic peptide, serum osmolality and haematocrit in the rat.

The present study documents daily rhythms of plasma atrial natriuretic peptide, serum osmolality and haematocrit in the rat. One-hundred and twenty-five Sprague-Dawley rats were used. They were bred under a cycle of 12 h light/12 h dark starting at 07.00 h. Fifty-three rats were decapitated between 09.00 and 16.00 h (study I) and 72 rats in groups of six were decapitated at 2-h intervals for a period of 24 h (study II). In study I, plasma atrial natriuretic peptide was 156 +/- 11 pg mg-1 (mean +/- SEM). In study II, atrial natriuretic peptide was at a control level from 08.00 to 18.00 h and then began to increase. At 22.00 h, atrial natriuretic peptide was 420 +/- 105 pg ml-1, which was significantly higher than at 08.00 h (P less than 0.05). The serum osmolality was over 300 mosmol kg-1 during the day. The highest mean osmolalities (315, 317, 312 mosmol kg-1) were found from 18.00 to 22.00 h. These were significantly different (P less than 0.05) from other groups during the day. The haematocrit was highest at 14.00 h (49.5 +/- 0.7%) and lowest at 24.00 h (43.6 +/- 0.8%) (P less than 0.05). In conclusion, we have shown that there are significant daily rhythms of plasma atrial natriuretic peptide, serum osmolality and haematocrit during a 24-h period and 12 h light/12 h dark cycle in the rat.

Adrenal cortex contributes to the regulation of NaCl-stimulated ANP release in the rat.

The present study documents the effects of hypophysectomy and the effects of dexamethasone substitution on the NaCl-stimulated release and on the basal secretion rates of ANP from the rat atria in vitro. We also measured the concentration of mRNA in the atria after hypophysectomy. Rats (n = 12) were subjected to hypophysectomy by a parapharyngeal approach. One group of rats (n = 6) received dexamethasone 0.2 mg s.c. daily for 4 weeks, while the other group was left unsubstituted. After 4 weeks, the atrial block (n = 10) was excised, placed in an organ bath (field stimulation 4 s-1, 20 V, 1 ms; resting tension = 5 mN) and superfused (7 ml min-1) either with a physiological buffer solution (295 mosmol kg-1) or with a hyperosmotic NaCl solution (330 mosmol kg-1). The atria from the hypophysectomized rats did not respond to the stimulus: the concentration of ANP in the 1-min samples of the perfusate was under 100 pg ml-1. Dexamethasone treatment significantly (P less than 0.05) increased the ANP concentration to a maximum of 165 +/- 17 (mean +/- SEM) pg ml-1 during the superfusion while the control concentration was 110 +/- 19 pg ml-1. The ANP mRNA/18 S RNA ratios did not differ between the atria of hypophysectomized and control rats. In conclusion, glucocorticoids are required in the stimulus-induced release of ANP and the impaired release of ANP after hypophysectomy does not depend on an impaired synthesis of ANP.