Conscious Calves Research Articles

Nitric oxide-related pancreatic endocrine responses to hyperglycaemia in the conscious calf.

Mean plasma insulin concentration was reduced and mean plasma glucose concentration increased following the administration of N-nitro-L-arginine methyl ester (L-NAME; 100 mumol kg-1 i.a.) in conscious calves given continuous infusions of exogenous glucose (30-60 mumol min-1 kg-1 i.v.). It is concluded that the rise in plasma insulin concentration which occurs in these animals in response to glucose is mediated, at least in part, by a nitric oxide-related factor (NOx).

Adrenal responses to the peptide PACAP in conscious functionally hypophysectomized calves

Intra-aortic infusions of pituitary adenylate cyclase-activating peptide-(1-38) (PACAP) produced a dose-related fall in aortic blood pressure over the range of 4-40 pmol.min-1.kg-1 in the presence of exogenous adrenocorticotropic hormone-(1-24) (ACTH, 2 ng.min-1.kg-1 i.v.; P < 0.01). At the higher dose there was a significant fall in adrenal vascular resistance in the absence, but not in the presence, of ACTH. PACAP also produced a dose-related increase in right adrenal cortisol output over the same range, which was significantly greater in the absence of exogenous ACTH (P < 0.01). At the higher dose, PACAP produced small but significant increases in adrenal epinephrine and norepinephrine output (P < 0.01) both in the presence and the absence of ACTH. There was also a small rise in Met5-enkephalin output, and corticotrophin-releasing factor (CRF) was released in the presence, but not in the absence, of ACTH. It is concluded that PACAP is capable of exerting potent steroidogenic and vasodilator effects in the adrenal gland in the normal conscious calf and of releasing significant amounts of catecholamines, enkephalins, and CRF from the adrenal medulla. These findings identify PACAP as a candidate neuromodulator in the adrenal gland in this species.

Adrenal cortical and medullary responses to acetylcholine and vasoactive intestinal peptide in conscious calves.

1. Adrenal responses to intra-aortic infusions of acetylcholine and vasoactive intestinal peptide (VIP) have been investigated in functionally hypophysectomized calves given exogenous adrenocorticotrophic hormone (ACTH, 2 ng min-1 kg-1 I.V.). 2. Infusions of VIP at a dose of 0.13 micrograms min-1 kg-1 caused a small, but significant increase in adrenaline and noradrenaline output which was, however, far below the level recorded previously in response to acetylcholine (0.7 micrograms min-1 kg-1). In contrast, these doses of the two agonists produced closely similar rises in adrenal cortisol output. 3. The steroidogenic effects of acetylcholine and VIP were found to be strictly additive and no evidence of potentiation was obtained in relation to either cortical or medullary responses or in the case of any of the cardiovascular responses which were monitored. 4. Intra-aortic infusions of VIP, at a dose which produced a substantial increase in adrenal steroidogenesis (0.065 micrograms min-1 kg-1), had no effect on the output of catecholamines, enkephalin-like immunoreactivity or corticotrophin-releasing factor, either in the presence or absence of acetylcholine. 5. It is concluded that VIP is unlikely to modulate adrenal medullary responses to muscarinic stimulation in this species as it has been claimed to do in the rat and does not potentiate adrenal steroidogenesis in response to acetylcholine as it does to ACTH.

Dissociation of hemodynamic and renal effects of i.v. alpha-hANF (99-126) in conscious calves.

The effects of i.v. human atrial natriuretic factor (alpha-hANF [99-126]) was investigated in 5 conscious calves (age 4 months, weight 94 +/- 14kg) with constant fluid (300 ml/h) and sodium intake over a period of 24 hours before and during the 3 hours of experimentation. We administered 200 micrograms, 400 micrograms and 800 micrograms ANF at hourly intervals. Immediately (+2 min) after the i.v. injection the peptide's serum-concentration rose from, basal 13 +/- 3 to 802 +/- 191 (200 micrograms), 1707 +/- 419 (400 micrograms) and 3483 +/- 878 pmol/l (800 micrograms) (p < 0.0001), respectively. Mean arterial pressure decreased from, basal, 108 +/- 18 to 85 +/- 17, 76 +/- 16 and 69 +/- 18 mmHg (p < 0.0001), and central venous pressure decreased from, basal, 5.1 +/- 3 to 1.2 +/- 1, 0.4 +/- 1 and 0.8 +/- 2 mmHg (p < 0.001). Heart rate increased from, basal, 66 +/- 5 to 84 +/- 23, 111 +/- 29 and 114 +/- 13 b/min (p < 0.001). Following the administration of ANF the urine volume decreased from, basal, 261 +/- 145 to 195 +/- 72, 121 +/- 41 and 102 +/- 33 ml/h (p < 0.0041). The urinary sodium excretion rates decreased from, basal, 50 +/- 17 to 30 +/- 13, 22 +/- 10 and 19 +/- 10 mmol/h (p < 0.007), and the potassium excretion rates decreased from, basal, 43 +/- 19 to 39 +/- 16, 26 +/- 11 and 23 +/- 10 mmol/h (p < 0.0047). Endogenous clearance of creatinine (basal: 182 +/- 30 ml/min) did not change (260 +/- 84, 218 +/- 66 and 224 +/- 76 ml/h) following i.v. ANF. Inspite of this marked fluid retention, the hematocrit, expressed as relative change, was increased by 7.5%, 11.8% and 10.4%. In 2 calves we additionally measured an increased whole blood and plasma density possibly indicating increased liquid permeation to an extravascular compartment. Thus, in calves the hemodynamic effects of ANF are comparable to those seen in man. However, the failure of ANF to stimulate diuresis and natriuresis indicates a dissociation of hemodynamic and renal effects of ANF in calves.

Effect of intraduodenal HCl and soybean extract on pancreatic juice secretion during atropinization and cold vagal blockade in calves.

In order to elucidate the mechanisms by which intraduodenal hydrochloric acid (HCl) and soybean extract influence exocrine pancreatic secretion in the young ruminant, we conducted experiments repeatedly on six conscious calves with and without blockade of the extrinsic and intrinsic neural pathways. In the absence of blockade, each of the two stimuli increased the juice volume, the HCl effect being far stronger than that of soybean extract. Intrinsic cholinergic blockade by atropinization blocked the stimulatory work of soybean extract on pancreatic secretion and on HCl-stimulated protein but had a weak effect on the amount of juice evoked by HCl. Temporary vagal blockade by chilling virtually abolished the excitatory effects of both soybean extract and HCl. With or without blockade, duodenal HCl resulted in a noteworthy increase in plasmal secretin and a slight increase in cholecystokinin (CCK). With alimentary proteins, acidification of the duodenum was responsible for both the composition and secretion of pancreatic juice in young calves, generally via neural pathways. Atropine-sensitive nerves of the pancreas totally regulate the intestinal phase of pancreatic juice secretion allied with intraduodenal protein, whereas HCl-dependent excitation of the exocrine pancreas takes place partly via atropine-resistant nerves. However, as hardly any pancreatic juice was secreted independently of the vagi, the vagi are deemed to govern all the postprandial regulatory mechanisms of the exocrine pancreas in the young calf.

The role of corticotrophin releasing factor in relation to the neural control of adrenal function in conscious calves.

1. Adrenal responses to intra-aortic infusions of pure synthetic ovine corticotrophin releasing factor (CRF), and to electrical stimulation of the preganglionic sympathetic innervation, have been investigated in functionally hypophysectomized conscious calves, in the presence and absence of a specific CRF antagonist. 2. CRF exerted a substantial steroidogenic effect on the adrenal gland of functionally hypophysectomized calves when infused intra-aortically at a dose (1.3 ng min-1 kg-1) below that which caused any fall in the arterial blood pressure. This response was significantly reduced, but not abolished by a concomitant infusion of CRF-antagonist into the aorta. 3. The steroidogenic effect of CRF was significantly reduced in the presence of exogenous adrenocorticotrophic hormone (ACTH) (2 ng min-1 kg-1, I.V.) and the surviving response was completely abolished by CRF-antagonist. 4. Stimulation of the peripheral end of the splanchnic nerve at 4 Hz in functionally hypophysectomized calves given exogenous ACTH produced a rise in mean adrenal output of the same order of magnitude as did exogenous CRF under the same conditions. The response to splanchnic nerve stimulation was apparently unaffected by CRF-antagonist although release of endogenous CRF from the gland was significantly increased thereby. 5. These results indicate that release of CRF from the adrenal gland during splanchnic nerve stimulation in the calf does not contribute significantly to the steroidogenic response thereto.

Muscarinic adrenal responses to acetylcholine in conscious calves.

1. Adrenal responses to intra-aortic infusions of acetylcholine (4.5 nmol min-1 kg-1 for 10 min) have been investigated in hypophysectomized conscious calves given exogenous adrenocorticotrophic hormone (ACTH) (2 ng min-1 kg-1 I.V.) in the presence and absence of hexamethonium. 2. Acetylcholine produced a significant increase in adrenal cortisol output and plasma cortisol concentration. In the absence of nicotinic blockade with hexamethonium this was apparently accounted for by an increase in adrenal ACTH presentation secondary to increased adrenal blood flow. However, administration of hexamethonium revealed a direct steroidogenic action of acetylcholine in the presence of exogenous ACTH. 3. Adrenal medullary responses to acetylcholine, including the release of catecholamines, enkephalins and corticotrophin-releasing factor (CRF), were not significantly reduced by nicotinic blockade. 4. It is concluded that both adrenal medullary and cortical responses to intraaortic infusions of acetylcholine at a low dose are mediated mainly by muscarinic receptors, as it has previously been shown that they are substantially reduced in the presence of atropine.

Acetylcholine-induced coronary vasoconstriction and vasodilation in tranquilized baboons.

To determine the effects of acetylcholine on the coronary bed in the baboon and whether the effects preceded or followed the action of acetylcholine on ventricular function, eight adult baboons (Papio anubis) were instrumented to measure left ventricular (LV) and mean arterial pressures, LV dP/dt, regional myocardial function, and coronary blood flow. Acetylcholine was injected locally through a catheter positioned in the coronary artery ostium using fluoroscopic guidance in intact sedated baboons. With heart rate held constant, intracoronary acetylcholine (0.5 micrograms/kg) reduced coronary blood flow by 82 +/- 4% from a baseline value of 34 +/- 4 ml/min without a significant change in mean arterial pressure and with a reduction in LV dP/dt of only 12 +/- 3%. The decrease in coronary blood flow occurred before either LV dP/dt or regional myocardial function fell in the region of the heart receiving acetylcholine. After the intense coronary constriction, a later phase characterized by dilation was observed. The changes in coronary blood flow with acetylcholine were unaffected by combined alpha- and beta-adrenoceptor blockades but were abolished by muscarinic blockade. Low doses of acetylcholine elicited only coronary vasodilation. All doses of acetylcholine, administered directly into the iliac artery, also elicited only iliac vasodilation. Intracoronary acetylcholine in conscious dogs also induced only coronary vasodilation, whereas in conscious calves at higher doses, initial vasoconstrictor responses were observed, which also preceded reductions in regional myocardial function. These results suggest that the controversy surrounding the effects of acetylcholine can be reconciled on the basis of species, vascular bed studied, and dose.(ABSTRACT TRUNCATED AT 250 WORDS)

Endocrine responses to intra-aortic infusions of acetylcholine in conscious calves.

1. Adrenal responses to intra-aortic infusions of acetylcholine (4.5 nmol min-1 kg-1 for 10 min) have been investigated in conscious, functionally hypophysectomized, 3- to 6-week-old calves, in the presence and absence of exogenous ACTH (2 ng min-1 kg-1, I.V.). 2. Acetylcholine produced a substantial fall in adrenal vascular resistance, which was significantly reduced in the presence of exogenous ACTH, while producing minimal changes in aortic blood pressure and heart rate. 3. There was also a significant rise in right adrenal cortisol output which was sufficient to produce a measurable rise in plasma cortisol concentration. The effect could be accounted for by the increase in adrenal ACTH presentation. It was abolished by pre-treatment with atropine (0.2 mg kg-1). A small but significant rise in aldosterone output during acetylcholine infusions was also abolished in the presence of ACTH. 4. Both adrenaline and noradrenaline were released during intra-aortic acetylcholine infusions and these responses were substantially reduced, but not abolished, by pre-treatment with atropine. 5. Acetylcholine also stimulated the release of corticotrophin-releasing factor (CRF) and [Met5]enkephalins from the gland. The output of CRF was enhanced and that of free [Met5]enkephalin was significantly reduced in the presence of exogenous ACTH. All these responses were largely, but not completely, suppressed by atropine. 6. Acetylcholine also promoted the release of the pancreatic hormones glucagon, insulin and pancreatic polypeptide (PP). The amounts of pancreatic glucagon and insulin that were released were highly dependent on the concentration of glucose in the circulating plasma and all these responses were abolished by atropine. 7. It is concluded that acetylcholine is capable of stimulating the release of a wide variety of agonists from the adrenal gland when infused intra-aortically at a dose of 4.5 nmol min-1 kg-1. The increase in cortisol output appears to be secondary to an increase in blood flow whereas the adrenal medullary responses are not, and appear to be due largely, but not entirely, to activation of muscarinic receptors.

The effect of splanchnic nerve stimulation on the uptake of atrial natriuretic peptide by the adrenal gland in conscious calves

A technique has been developed with which it has been possible to quantify the output of a wide variety of agonists including catecholamines, glucocorticoids, mineralocorticoids, enkephalins and various peptides, from the adrenal gland in the conscious unrestrained calf; also to investigate responses to electrical stimulation of the peripheral end of the splanchnic nerve below any behavioural threshold. In the present study this methodology has been employed to investigate the extent to which stimulation of the splanchnic sympathetic innervation affects adrenal handling of atrial natriuretic peptide as this peptide has been identified within the adrenal medulla in this species. Stimulation of the splanchnic nerve at frequencies which raised the concentration of atrial natriuretic peptide like-immunoreactivity (ANP) by 25% led to an abrupt increase in the uptake of the peptide by the right adrenal gland by about 250%. During nerve stimulation more than 20% of the ANP that was estimated to be presented to the gland was taken up, by comparison with less than 13% of the amount presented which was taken up before and after stimulation. These results suggest that stimulation of the splanchnic nerve may specifically enhance the uptake of ANP by the adrenal gland and represent the first report of such a mechanism in respect of any biologically active peptide so far as we are aware.

The effect of changes in adrenal blood flow on adrenal cortical responses to adrenocorticotrophin in conscious calves.

1. The effect of varying adrenal blood flow on the rate at which it was estimated that adrenocorticotrophin (ACTH) was presented to the adrenal gland was related to right adrenal cortisol output in conscious calves fitted with 'adrenal clamps'. 2. Intra-aortic infusions of endothelin at either 15.0 or 7.5 pmol min-1 kg-1 produced a substantial fall in right adrenal blood flow which was dose-related over this range. There was an associated fall in right adrenal cortisol output and cortisol output was linearly related to estimated ACTH presentation to the gland over the whole range investigated. The changes in adrenal cortisol output were reflected by changes in the concentration of cortisol in the peripheral plasma, which could be attributed entirely to the fluctuations in adrenal cortisol output. 3. It is concluded that delivery of ACTH to the adrenal gland is flow dependent over the physiological range in these animals and that changes in adrenal cortical blood flow can therefore be expected to result in changes in adrenal output due to variations in the presentation rate of ACTH.

Release of adrenocorticotrophin from the adrenal gland in the conscious calf.

1. The effect of stimulation of the splanchnic nerve on the output of ACTH-related peptides from the adrenal gland has been investigated in conscious, functionally hypophysectomized calves, previously fitted with an 'adrenal clamp'. 2. Stimulation of the splanchnic nerve elicited a small, but statistically significant, increase in the output of ACTH-like immunoreactivity at each frequency tested. This response was frequency-dependent over the range 40-70 Hz when stimulating intermittently for 1 s at 10 s intervals and was potentiated by stimulating intermittently. Thus, the average mean output during stimulation in burst at 70 Hz (25 +/- 5 fmol min-1 kg-1) was significantly higher than the corresponding value during continuous stimulation at 7 Hz (6 +/- 1 fmol min-1 kg-1; P less than 0.01) even though the total number of impulses delivered was identical in each case. 3. There was also a small but significant rise in the output of cortisol from the gland with intermittent stimulation, which was linearly related to the output of ACTH-like immunoreactivity at the lower frequencies (4 and 7 Hz). 4. Separation of the ACTH-related peptides which were extracted from the adrenal effluent plasma of these animals during splanchnic nerve stimulation revealed the existence of two clear forms: ACTH (1-39) accounted for about 60% of total ACTH immunoreactivity and pro-opiomelanocortin (POMC) for about 30%. 5. It is concluded that small amounts of ACTH are released within the adrenal gland during splanchnic nerve stimulation in the functionally hypophysectomized calf and that this may possibly contribute towards the steroidogenic effect of stimulating the splanchnic nerve.

Beta-receptors and adenylate cyclase: comparison of nonischemic, ischemic, and postmortem tissue

We compared the effects of myocardial ischemia and postmortem changes on beta-adrenergic receptors and their coupling to adenylate cyclase activity. The effects of 1 h of left circumflex coronary artery occlusion were examined in eight conscious calves, which were then anesthetized with pentobarbital sodium, and the left ventricle was divided into nonischemic and ischemic regions. A crude membrane fraction was prepared from each region and from the nonischemic tissue 1 h postmortem. beta-Adrenergic receptor density increased (152 +/- 55%) and decreases in basal (-21 +/- 6.1%), isoproterenol-stimulated (-25 +/- 8.0%), 5'-guanylylimidodiphosphate [Gpp(NH)p]-stimulated (-17 +/- 5.8%), fluoride-stimulated (-26 +/- 5.8%), and forskolin-stimulated (-31 +/- 8.4%) adenylate cyclase activities were observed in the ischemic myocardium compared with nonischemic myocardium. Similarly, in postmortem samples, beta-adrenergic receptor density rose 58 +/- 16%, whereas decreases in basal (-48 +/- 8.7%), isoproterenol-stimulated (-61 +/- 7.8%), Gpp(NH)p-stimulated (-58 +/- 7.0%), fluoride-stimulated (-64 +/- 6.1%), and forskolin-stimulated (-52 +/- 6.2%) adenylate cyclase activities were observed. Agonist-binding competition curves with isoproterenol were shifted, indicating that beta-adrenergic receptors were binding agonists with low affinity in both the ischemic and postmortem myocardium. The marked, but directionally opposite, changes in receptor density and adenylate cyclase that occur postmortem indicate the importance of prompt processing of tissues. The striking similarity in response of beta-adrenergic receptor agonist and antagonist binding and adenylate cyclase activity in ischemic and postmortem tissue raises the speculation that similar mechanisms may operate under both conditions.

Adrenal responses to splanchnic nerve stimulation in conscious calves given naloxone.

1. The effects of stimulating the peripheral end of the right splanchnic nerve in the presence of naloxone (2 mg kg-1) have been investigated in conscious 3 to 6-week-old calves. 2. Mean aortic blood pressure rose to significantly higher levels during splanchnic stimulation in bursts at 40 Hz for 1 s at 10 s intervals than it did during stimulation at the corresponding continuous frequency (4 Hz). Furthermore, naloxone significantly reduced the fall in mean vascular resistance in response to both patterns of stimulation. 3. The output of catecholamines from the adrenal gland, together with the proportion of noradrenaline released, was significantly enhanced by stimulating the splanchnic nerves in bursts in animals pre-treated with naloxone and the proportion of noradrenaline released also increased. In both cases the output of adrenaline and noradrenaline was within the same range as that reported previously in normal control animals. 4. Naloxone significantly increased the amounts of enkephalin-like immunoreactivity and corticotrophin-releasing factor (CRF)-like immunoreactivity released from the adrenal gland in response to splanchnic nerve stimulation and raised the proportion of total to free met5-enkephalin that was secreted. 5. Naloxone also inhibited the rise in plasma adrenocorticotrophic hormone (ACTH) concentration during continuous stimulation at 4 Hz, but not during stimulation at 40 Hz in bursts. Under these latter conditions the output of cortisol apparently directly from the adrenal gland was inhibited. The finding that splanchnic nerve stimulation can potentiate the output of cortisol in response to ACTH was confirmed. 6. These results provide evidence that release of enkephalins and of CRF from the adrenal is inhibited by activating opioid receptors within the gland itself.

Species differences in regulation of alpha-adrenergic receptor function

The effects of alpha-adrenergic receptor stimulation with norepinephrine and phenylephrine after beta-adrenergic and muscarinic receptor blockades were compared in conscious baboons, calves, dogs, and rats in which left ventricular (LV) pressure. LV maximum rate of pressure development (dP/dt), and heart rate were measured. Autonomic receptor density was examined in crude sarcolemmal preparations from the hearts. The major physiological differences were observed in rats, where alpha 1-adrenergic receptor stimulation resulted in the greatest (P less than 0.05) increases in LV dP/dt (29 +/- 2%) in response to phenylephrine, 5.0 micrograms/kg, in comparison with responses in dogs (12 +/- 4%), calves (3 +/- 3%), and baboons (1 +/- 2%). This was associated with the greatest (P less than 0.05) alpha 1-adrenergic receptor density in the rat heart (73 +/- 5 fmol/mg) compared with values in the baboon (6 +/- 1 fmol/mg), calf (21 +/- 3 fmol/mg), or dog (10 +/- 3 fmol/mg) myocardium. Thus there are major differences among mammalian species in alpha-adrenergic receptor density and physiological responsiveness to alpha-adrenergic agonists.

Regulation of bronchomotor tone in conscious calves.

The purpose of this study was to investigate the effects of some alpha and beta sympathomimetic and sympatholytic drugs on respiratory impedance in healthy conscious calves. Ten Friesian calves were investigated in this study. The forced oscillation technique was used to measure the resistance (Rrs) and the reactance (Xrs) of the respiratory system at frequencies ranging from 4 to 26 Hz. Isoprenaline (1 microgram/kg i.v.), propranolol (3 micrograms/kg i.v.), noradrenaline (2 micrograms/kg i.v.), xylazine (20 micrograms/kg i.v.) and yohimbine (0.25 mg/kg i.v.) were were administered. Isoprenaline induced a significant decrease of Rrs. An increase of Rrs after administration of propranolol was observed but without any change of the frequency dependence of Rrs. A small increase in the resonant frequency was also recorded. A decrease of Rrs was recorded after yohimbine injection. Noradrenaline and xylazine administration increased the resistances and the resonant frequency and induced a negative frequency dependence of Rrs. These results suggest that (1) the major effects of beta adrenergic drugs are on the central airways, (2) the alpha adrenergic system may play a role on the regulation of bronchomotor tone in calves, (3) the effects of alpha adrenergic drugs are on both central and peripheral airways and (4) the forced oscillation technique allows the differentiation of calibre changes occurring in small and large airways.

Adrenal responses to calcitonin gene‐related peptide in conscious hypophysectomized calves.

1. Right adrenal and various cardiovascular responses to an intra-aortic infusion of calcitonin gene-related peptide (CGRP; 4 micrograms min-1) have been investigated in the presence and absence of exogenous adrenocorticotrophin ACTH1-24 (2 or 5 ng min-1 kg-1, I.V.). The adrenal clamp technique was employed in conscious calves in which the pituitary stalk had been cauterized 3-7 days previously. 2. At the higher dose (5 ng min-1 kg-1) the I.V. infusion of ACTH raised mean plasma ACTH concentration by about 1000 pg ml-1 and mean right adrenal cortisol output by about 750 ng min-1 kg-1. Under these conditions the intra-aortic infusion of CGRP had no apparent effect on adrenal cortisol output by about 750 ng min-1 kg-1. Under these conditions the intra-aortic infusion of CGRP had no apparent effect on adrenal function, other than to produce moderate adrenal vasodilatation. In contrast, in the absence of exogenous ACTH, the same dose of CGRP produced a substantial rise in cortisol output, which rose steadily to a peak mean value of 409 +/- 31 pg min-1 kg-1 at 10 min. It also significantly inhibited the release of free, but not of total, met5-enkephalin-like immunoreactivity from the gland (P less than 0.001) together with a significantly greater fall in adrenal vascular resistance (P less than 0.001). 3. At the lower dose of ACTH (2 ng min-1 kg-1, I.V.) CGRP raised mean plasma cortisol output from 314 +/- 31 to 486 +/- 44 ng min-1 kg-1 (P less than 0.01) and this effect was not attributable to an increase in the adrenal presentation rate of ACTH. 4. It is concluded that this peptide exerts a steroidogenic action on the adrenal cortex which is manifest in the absence of exogenous ACTH in the functionally hypophysectomized calf.

Alpha-adrenergic vasoconstriction and receptor subtypes in large coronary arteries of calves.

We investigated alpha-adrenoceptor subtype distribution in large coronary arteries from both functional and biochemical perspectives. The effects of intracoronary administration of the selective alpha 1-adrenoceptor agonist phenylephrine (0.1-5.0 micrograms/kg ic), of the selective alpha 2-adrenoceptor agonist B-HT 920 (0.1-5.0 micrograms/kg ic), and of the mixed alpha 1 + 2-adrenoceptor agonist norepinephrine (0.01-0.5 micrograms/kg ic) were examined on measurements of left circumflex coronary artery diameter in conscious calves. After beta-adrenergic blockade, equivalent reductions in large coronary artery diameter were observed with phenylephrine (-6.9 +/- 0.7%), B-HT (-5.9 +/- 0.5%), and norepinephrine (-6.0 +/- 0.4%). Phenylephrine-induced constrictions were abolished by prazosin, an alpha 1-selective antagonist, but unaffected by rauwolscine, an alpha 2-selective antagonist. Conversely, the B-HT-induced constriction was abolished by rauwolscine but unaffected by prazosin. Coronary constriction with norepinephrine was attenuated with either prazosin or rauwolscine and abolished by the two antagonists combined. Ligand-binding studies in which [3H]prazosin and [3H]rauwolscine and sarcolemmal membranes were used revealed an alpha 1-adrenoceptor density of 15 +/- 3.1 fmol/mg protein with a dissociation constant (KD) of 0.7 +/- 0.2 nM and an alpha 2-adrenoceptor density of 68 +/- 5.1 fmol/mg protein, with a KD of 7.4 +/- 1.2 nM. Thus large coronary arteries of the calf contain both alpha 1- and alpha 2-adrenoceptor subtypes, each of which elicits constriction of the large coronary artery in the conscious animal.

Increase in plasma concentrations of atrial natriuretic peptide during infusion of hypertonic saline in conscious newborn calves.

Plasma concentrations of atrial natriuretic peptide (ANP), plasma renin activity (PRA), plasma concentrations of aldosterone, urine flow rate and sodium and potassium excretion were studied in two groups of four conscious 3-day-old male calves, infused with hypertonic saline or vehicle. Hypertonic saline infusion (20 mmol NaCl/kg body weight) was accompanied by a progressive rise in plasma concentrations of ANP (from 16.5 +/- 0.2 pmol/l at time 0 to 29.3 +/- 3.0 pmol/l at 30 min; P less than 0.05) and by a gradual decrease in PRA (from 1.61 +/- 0.23 nmol angiotensin I/l per h at time 0 to 0.54 +/- 15 nmol angiotensin I/l per h at 90 min; P less than 0.05); there was no change in the plasma concentration of aldosterone. Within the first 2 h of the 24-h urine collection period there was a marked rise in urine flow rate and sodium excretion in treated calves when compared with control animals (66.0 +/- 8.3 vs 15.9 +/- 1.2 ml/kg body weight per 2 h (P less than 0.05) and 6.7 +/- 1.3 vs 0.4 +/- 0.02 mmol/kg body weight per 2 h (P less than 0.01) respectively). During the following 22 h, urinary water and sodium excretion remained at significantly high levels. Thus, in the conscious newborn calf, changes in plasma ANP levels and urinary water and sodium excretion during hypertonic saline infusion are compatible with the hypothesis that endogenous ANP participates, at least in part, in the immediate diuretic and natriuretic renal response induced by the sodium overload.

Secretion of corticotrophin releasing factor from the adrenal during splanchnic nerve stimulation in conscious calves.

1. The output of corticotrophin releasing factor-like immunoreactivity (CRF) from the adrenal gland has been investigated using the 'adrenal clamp' technique in conscious calves. 2. Stimulation of the peripheral end of the splanchnic nerve for 10 min increased the mean output of CRF progressively, so that it had risen by about twentyfold, to a peak incremental value of 24 +/- 4 pmol min-1 kg-1 at 10 min. This response was significantly increased by stimulating in bursts at 40 Hz for 1 s at 10 s intervals, which raised the mean CRF output by 44 +/- 7 pmol min-1 kg-1 at 10 min (P less than 0.05). 3. The mean output of adrenaline and noradrenaline rose more abruptly in response to splanchnic nerve stimulation with peak incremental values realized within 2.5 min. However, the ratios of adrenal CRF to catecholamine output were closely similar during the later stages of stimulation (7.5-10 min). There was a similarly abrupt rise in adrenal cortisol output in response to splanchnic nerve stimulation which was, nevertheless, linearly related to arterial plasma ACTH concentration throughout. 4. In hypophysectomized calves, administration of adrenocorticotrophic hormone (ACTH1-24) at a dose of 5 ng min-1 kg-1 reduced the output of adrenal CRF in response to splanchnic nerve stimulation by about 50% (P less than 0.05). 5. CRF isolated from adrenal venous effluent plasma, collected both at rest and during splanchnic nerve stimulation, was separated by reverse-phase high-pressure liquid chromatography and found to elute in a position identical to that of human 41CRF. This suggests that adrenal CRF is structurally closely similar to its pituitary counterpart.