Pharmacological evidence for capacitative Ca(2+) entry in cannulated and pressurized skeletal muscle arterioles.

Arteriolar myogenic tone shows a marked dependency on extracellular Ca(2+). The contribution played by mechanisms such as intracellular Ca(2+) release and capacitative entry, however, are less certain. The present studies aimed to demonstrate functional evidence for involvement of such mechanisms in myogenic tone and reactivity. Single cremaster arterioles were denuded of endothelium, pressurized under no-flow conditions and loaded with fura 2-AM for measurement of changes in intracellular Ca(2+) [Ca(2+)](i). The cell permeable, putative, IP(3) receptor antagonist 2APB (2 aminoethoxydiphenyl borate) was used to determine the possible role of IP(3) receptor-mediated mechanisms in arteriolar myogenic tone and reactivity. Arterioles dilated in response to increasing concentrations of 2APB (1 - 300 microM) without a concomitant change in global [Ca(2+)](i). Also 2APB (50 microM) completely inhibited the myogenic constriction in response to a step change in luminal pressure (50 - 120 mmHg) with no apparent effect on pressure-mediated increases in [Ca(2+)](i). 2APB markedly attenuated the constrictor response and [Ca(2+)](i) increase stimulated by phenylephrine but not KCl. Capacitative Ca(2+) influx in arterioles was demonstrated either by re-addition of extracellular [Ca(2+)] following pre-treatment with 1 or 10 microM nifedipine in Ca(2+) free buffer or exposure of vessels to thapsigargin (1 microM) to induce store depletion. In both cases 2APB inhibited the increase in [Ca(2+)](i). Capacitative Ca(2+) entry showed an inverse relationship with intraluminal pressure over the range 10 - 120 mmHg. Consistent with an effect on a Ca(2+) entry pathway, 2APB had no effect on intracellular (caffeine releasable) Ca(2+) stores while decreasing the rate of Mn(2+) quench of fura 2 fluorescence. The results provide functional evidence for capacitative Ca(2+) entry in intact arteriolar smooth muscle. The effectiveness of 2APB in inhibiting both non-voltage gated Ca(2+) entry and responsiveness to an acute pressure step is consistent with the involvement of an axis involving IP(3)-mediated and or capacitative Ca(2+) entry mechanisms in myogenic reactivity. Given the lack of effect of 2APB on pressure-induced changes in global [Ca(2+)](i) it is suggested that such mechanisms participate on a localized level to couple the myogenic stimulus to contraction.

Invited review: arteriolar smooth muscle mechanotransduction: Ca(2+) signaling pathways underlying myogenic reactivity.

The smooth muscle of arterioles responds to an increase in intraluminal pressure with vasoconstriction and with vasodilation when pressure is decreased. Such myogenic vasoconstriction provides a level of basal tone that enables arterioles to appropriately adjust diameter in response to neurohumoral stimuli. Key in this process of mechanotransduction is the role of changes in intracellular Ca(2+). However, it is becoming clear that considerable complexity exists in the spatiotemporal characteristics of the Ca(2+) signal and that changes in intracellular Ca(2+) may play roles other than direct effects on the contractile process via activation of myosin light-chain phosphorylation. The involvement of Ca(2+) may extend to modulation of ion channels and release of Ca(2+) from the sarcoplasmic reticulum, alterations in Ca(2+) sensitivity, and coupling between cells within the vessel wall. The purpose of this brief review is to summarize the current literature relating to Ca(2+) and the arteriolar myogenic response. Consideration is given to coupling of Ca(2+) changes to the mechanical stimuli, sources of Ca(2+), involvement of ion channels, and spatiotemporal aspects of intracellular Ca(2+) signaling.

Effects of mibefradil and nifedipine on arteriolar myogenic responsiveness and intracellular Ca(2+).

1. Ca(2+) entry mechanisms underlying spontaneous arteriolar tone and acute myogenic reactivity remain uncertain. These studies aimed to compare the effects of nifedipine and the putative T-channel blocker, mibefradil, on arteriolar myogenic responsiveness and intracellular Ca(2+) (Ca(2+)(i)). 2. First order cremaster muscle arterioles (1A) were isolated from rats, cannulated, pressurized to 70 mmHg in the absence of intraluminal flow, and mechanical responses studied by video microscopy. The Ca(2+)(i) was measured using fluorescence imaging of Fura 2 loaded arterioles. 3. Both nifedipine and mibefradil showed dose-dependent inhibition of spontaneous myogenic tone (at 70 mmHg; pEC(50) 7.04+/-0.17 vs 6.65+/-0.20 respectively, n=6 for both, n.s.) and KCl-induced vasoconstriction (at 70 mmHg; pEC(50) 6.93+/-0. 38 vs 6.45+/-0.27 respectively, n=6 for both, n.s.). 4. In arterioles maintained at 50 mmHg, nifedipine (10(-7) and 10(-5) M) caused a concentration dependent reduction in Ca(2+)(i), however, mibefradil (10(-7) and 10(-5) M) had no effect. Furthermore nifedipine significantly attenuated the increase in Ca(2+)(i) associated with an acute pressure step (50 - 120 mmHg) whereas mibefradil was considerably less effective. 5. Mibefradil (10(-7) M) significantly attenuated contractile responses to 60 mM KCl without altering the KCl-induced increase in Ca(2+)(i), in contrast to nifedipine (10(-7) M) which reduced both Ca(2+)(i) and contraction. 6. Membrane potential of arterioles with spontaneous myogenic tone (70 mmHg) was -41.5+/-1. 0 mV. Nifedipine (10(-7) or 10(-5) M) had no effect on membrane potential, however mibefradil (10(-5) M) caused significant depolarization. 7. In summary, both mibefradil and nifedipine inhibit arteriolar spontaneous tone and acute myogenic reactivity. While there may be overlap in the mechanisms by which these agents inhibit tone, differences in effects on membrane potential and intracellular Ca(2+) levels suggest mibefradil exhibits actions other than blockade of Ca(2+) entry in skeletal muscle arterioles.

Transient increases in diameter and [Ca(2+)](i) are not obligatory for myogenic constriction.

Studies were performed to determine the significance of temporal variation in vascular smooth muscle Ca(2+) signaling during acute arteriolar myogenic constriction and, in particular, the importance of the stretch-induced intracellular Ca(2+) concentration ([Ca(2+)](i)) transient in attaining a steady-state mechanical response. Rat cremaster arterioles (diameter approximately 100 microm) were dissected from surrounding tissues, and vessel segments were pressurized in the absence of intraluminal flow. For [Ca(2+)](i) measurements, vessels were loaded with fura 2 and fluorescence emitted by excitation at 340 and 380 nm was measured using video-based image analysis. Ca(2+) and diameter responses were examined after increases in intravascular pressure were applied as an acute step increase or a ramp function. Additional studies examined the effect of longitudinal vessel stretch on [Ca(2+)](i) and arteriolar diameter. Step increase in intraluminal pressure (from 50 to 120 mmHg) caused biphasic change in [Ca(2+)](i) and diameter. [Ca(2+)](i) transiently increased to 114.0 +/- 2.0% of basal levels and subsequently declined to 106.7 +/- 4.4% at steady state. Diameter initially distended to 125.4 +/- 2.1% of basal levels before constricting to 71.1 +/- 1.2%. In contrast, when the same pressure increase was applied as a ramp function (over 5 min) transient vessel distension and transient increase in [Ca(2+)](i) were prevented, yet at steady state vessels constricted to 71.3 +/- 2.5%. Longitudinal stretch resulted in a large [Ca(2+)](i) transient (158 +/- 19% of basal) that returned to baseline despite maintenance of the stretch stimulus. The data demonstrate that the initial vessel distension (reflecting myocyte stretch) and associated global [Ca(2+)](i) transient are not obligatory for myogenic contraction. Thus, although arteriolar smooth muscle cells are responsive to acute stretch, the resulting changes in myogenic tone may be more closely related to other mechanical variables such as wall tension.

Myogenic reactivity of rat epineurial arterioles: potential role in local vasoregulatory events.

Local control of neural blood flow is considered to reside in innervation of epineurial and endoneurial arterioles rather than in intrinsic autoregulatory mechanisms. With the use of an isolated vessel preparation and an in vivo approach, the present studies examined intrinsic vasomotor responsiveness of epineurial arterioles. Segments of epineurial arterioles, cannulated on glass micropipettes (40 micrometers) and pressurized in the absence of intraluminal flow, showed sustained pressure-dependent (30-90 mmHg) vasoconstriction and acute myogenic reactivity. Myogenic tone was unaffected by phentolamine (10(-6) M). Removal of extracellular Ca(2+) resulted in loss of spontaneous tone and passive behavior. Concentration-response curves for norepinephrine (10(-9)-3 x 10(-6) M) and relaxation to both acetylcholine (10(-8)-10(-5) M) and adenosine (10(-8)-10(-4) M) were obtained. Acetylcholine dilator responses were inhibited by N(G)-nitro-L-arginine methyl ester. Epineurial blood flow was measured in vivo using a laser-Doppler flow probe. Blood flow declined over a 2-h period after surgery, and during this time preparations developed responsiveness to the dilator acetylcholine. Phentolamine blocked vasoconstrictor responses to exogenous norepinephrine but only partially reversed the in vivo baseline tone. The time-dependent decline in epineurial blood flow was observed despite the presence of tetrodotoxin (1 microM), further confirming that tone was predominantly caused by myogenic rather than neurogenic mechanisms. It is concluded that because epineurial arterioles exhibit intrinsic myogenic reactivity, they have the potential to participate in local regulation of neural hemodynamics independently of their own innervation.

Ontogeny of hormonal and excretory function of the meso- and metanephros in the ovine fetus.

Using reverse-transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry we investigated the ontogeny of renin, angiotensinogen and angiotensin converting enzyme (ACE) in the mesonephros at 27 and 41 days of gestation, and the metanephros at 41 and 64 days of gestation in ovine fetuses (term is 145 to 150 days). The volume and composition of fetal urine, stored as allantoic fluid were measured in 12 fetuses at 27 days, and 13 fetuses at 41 days. Renin, angiotensinogen and ACE were identified in both meso- and metanephroi at 41 days but not in the mesonephros at 27 to 30 days. Allantoic fluid volumes were 21 +/- 3 and 45 +/- 5 ml at 27 to 30 days and 41 days, respectively. This fluid was significantly different in composition to that of amniotic fluid or maternal plasma. The results suggest that the mesonephros can substantially modify its glomerular filtrate by 27 days of gestation, and can produce local angiotensin II by 41 days.

Development of the spleen as a red blood cell reservoir in lambs.

In adult sheep and other animals the spleen is a reservoir of red blood cells (RBCs), which can be mobilized by adrenaline-induced splenic contraction. Adrenaline does not increase the haematocrit in the ovine fetus. This study examined the development of a releasable store of RBCs in the spleen of lambs during the neonatal period. Spleen and body weights were measured in 126 fetuses and lambs, ranging in age from 60 days' gestation to 94 days post partum. Spleen weight, as a percentage of body weight, stabilized at 0.56 +/- 0.02%, 20 days post partum. Adrenaline infusion (5 mg kg-1 min-1) was associated with a mean increase in arterial haematocrit (Hct) of 7.6 and 5.8 Hct % in lambs at 20-24 and 50-52 days of age respectively (n = 4 at each age). The spleen in lambs has become a significant reservoir of RBCs by 20-24 days of age and may contain reserves equivalent to those in adult sheep. This splenic reserve is developed before the post-natal drop in haematocrit occurs.

Cardiovascular, hormonal, and metabolic responses to severe prolonged hemorrhage in adult sheep.

Over a 54-hour period, blood was removed from 8 adult sheep (body weight, 38.1 +/- 0.5 kg, mean +/- SEM) in 9 episodes, 5 on day 1, 3 on day 2, and 1 on day 3. Cumulative blood loss was 1,630 +/- 63, 2,380 +/- 71, and 2,693 +/- 69 ml on days 1, 2, and 3, respectively. Blood samples (20 ml) were collected from 5 control ewes (33.8 +/- 2.8 kg) at equivalent times. Over the first day, mean arterial blood pressure decreased in the hemorrhaged sheep from 101 +/- 2 mm of Hg to 76 +/- 5 mm of Hg, but returned to control values by the beginning of the second day and, thereafter, was not different from control values. Heart rate was increased after the first hemorrhage episode and remained high throughout the entire protocol. Over the entire period, there were statistically significant decreases in hematocrit, plasma osmolality, sodium, total calcium (P < 0.001), potassium, and chloride values (P < 0.05). There was no change in plasma phosphate, bicarbonate, creatinine, or magnesium concentrations and an increase in plasma urea nitrogen (P < 0.001) concentrations. Plasma arginine vasopressin concentration was increased significantly (P < 0.001) over the entire period. Plasma ACTH concentration was significantly (P < 0.05) increased over time, but only some values on day 1 were significantly outside the normal range of the control group data. Because of wide variation between sheep, the group data for aldosterone were not significantly different from control values.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrial natriuretic peptide receptors are present and functional by midgestation in fetal sheep.

This study examined the ontogeny of the mRNA for three atrial natriuretic peptide (ANP) receptors in the ovine fetal kidney and the effect of systemic ANP infusion in the very immature ovine fetus. mRNA was isolated from the kidneys of 60-, 100-, and 140-day fetuses (n = 4 at each age). Northern blots [5 micrograms poly(A)+ RNA per track] were probed for the guanylate cyclase (GC)-A, GC-B, and clearance receptors, using beta-actin as a control for variations in loading. The results were quantitated using laser densitometry. Levels of clearance receptor mRNA were significantly higher in 140-day than 60-day fetal kidneys (P < 0.05), whereas levels of mRNA for the GC-A and GC-B receptors remained steady. We propose that binding of ANP to an increased number of C receptors in the late-gestation fetal kidney could explain the previously documented increase in total ANP receptor number in late-gestation ovine kidneys without increased ANP biological activity. Systemic ANP infusion into four fetuses of approximately 74 days gestation resulted in a natriuresis and diuresis, indicating the presence of functional ANP receptors in the ovine kidney early in gestation.

Blood volume measurements in the neonatal lamb: validation of a method using [51Cr]-labelled red cells.

1. The reproducibility of blood volume measurements, using [51Cr]-labelled red blood cells (RBC) was tested in three lambs, 79-89 days of age, and the accuracy was tested in eight lambs, 83 +/- 0.4 days of age, in which blood volume was measured simultaneously by [51Cr]-RBC and [125I]-gamma globulin plus haematocrit (hct). 2. The blood volumes (mean +/- s.e.) of the three lambs, on four occasions, were 64.5 +/- 2.5, 68.8 +/- 2.9 and 63.9 +/- 3.7 mL/kg, respectively, and the coefficients of variation were 7.8, 8.6 and 11.6%, respectively. 3. The mean blood volume of eight lambs was 60.9 +/- 3.3 mL/kg by [51Cr]-RBC, and 60.8 +/- 2.4 mL/kg from plasma volume and hct. These were not statistically different. 4. Arterial hct was 31.2 +/- 0.5%, which was not statistically different from the whole body hct of 31.2 +/- 2.6%. 5. It is concluded that [51Cr]-RBC can be used to measure accurate and reproducible blood volumes in lambs.

Effect of maternal glucocorticoid treatment on fetal fluids in sheep at 0.4 gestation.

Treatment of nine pregnant Merino ewes (64.0 +/- 0.4 days of gestation) with dexamethasone (D; 0.76 mg/h for 48 h) resulted in significant alterations in fetal fluids compared with eight saline-infused control animals (S; 63.0 +/- 0.9 days). There was a substantial increase in allantoic fluid volume (177 +/- 18 ml, D vs. 31 +/- 6, S) but no change in amniotic fluid volume (248 +/- 12 ml, D; 305 +/- 24, S). For allantoic fluid there was a significant decrease in osmolality (213 +/- 4 mosmol/kg water, D; 230 +/- 5, S) and alterations in composition. Amniotic fluid osmolality was unchanged (292 +/- 2 mosmol/kg water, D; 293 +/- 1, S), but amniotic fluid composition was affected. In four fetuses in which bladder and amniotic cannulas were inserted at gestational age 68-75 days, fetal urine flow rate increased from a mean of 4.1 +/- 1.1 to 13.8 +/- 2.6 ml/h after 24 h and 11.8 +/- 3.0 ml/h at 48 h for a similar maternal D infusion, whereas no such increase occurred in four control fetuses. All the fetal urine voided during a 3.5- to 4-h infusion of 51Cr-labeled EDTA into the fetal bladder was directed to the allantois. The results suggest that the increase in allantoic fluid volume resulted from increased fetal urine output into the allantoic compartment, although the composition of the excess allantoic fluid differed substantially from that of fetal urine. There was a greater incidence of abnormal cotyledons in the D-infused ewes.(ABSTRACT TRUNCATED AT 250 WORDS)

Intracisternal naloxone and cardiac nerve blockade prevent vasodilatation during simulated haemorrhage in awake rabbits.

1. Acute haemorrhage was simulated in five unanaesthetized rabbits, by inflating a cuff on the inferior vena cava so that cardiac output fell by 8.3% of its resting level per minute. Simulated haemorrhage was performed after sham treatment, after graded doses of intravenous and intracisternal naloxone, and after cardiac nerve blockade with intrapericardial procaine. 2. After sham treatment, the haemodynamic response to simulated haemorrhage was biphasic. During the first phase, systemic vascular conductance fell steadily, heart rate rose steadily, and arterial pressure fell only slightly. A second decompensatory phase began abruptly when cardiac output had fallen to approximately 55% of its resting level. Vascular conductance rose steeply, heart rate fell slowly, and arterial pressure fell precipitately. 3. Treatment with naloxone (intravenous, 0.04-0.4 mg kg-1; intracisternal, 0.2-2 micrograms kg-1) did not affect either phase of the haemodynamic response to simulated haemorrhage. 4. After treatment with larger doses of naloxone (intravenous, 4-8 mg kg-1; intracisternal, 4-69 micrograms kg-1), the first phase was unaffected, but the second phase no longer occurred. Throughout simulated haemorrhage, systemic vascular conductance fell steadily, heart rate rose, and arterial pressure was well maintained. The dose of intracisternal naloxone which prevented the second phase was 90-900 times less than the corresponding intravenous dose. The second phase was also prevented by cardiac nerve blockade. 5. We conclude that an endogenous opiate mechanism is responsible for the haemodynamic decompensation that occurs when cardiac output falls to a critical level. The mechanism is located within the central nervous system. It is triggered by a signal from the heart.

Simulation of acute haemorrhage in unanaesthetized rabbits.

1. We have shown that it is feasible to match the linear rate of fall of cardiac output that occurs during haemorrhage at 2.7 ml/kg per min in unanaesthetized rabbits by constricting the thoracic inferior vena cava so as to decrease venous return. 2. The changes in systemic vascular resistance, arterial pressure and heart rate that occurred during haemorrhage were mimicked by simulated haemorrhage. They were reproducible when simulated haemorrhage was performed three times at 90 min intervals, and when it was repeated four times over 12 days. 3. Simulated haemorrhage caused rises in plasma renin activity (PRA) and plasma arginine vasopressin concentration (AVP) that were similar to those reported after haemorrhage. The response of PRA was unaffected by repeated simulated haemorrhage, but the response of AVP was less on the third occasion. 4. When the shed blood was re-infused after haemorrhage, cardiac output tended to remain low and systemic vascular resistance high. After simulated haemorrhage, all haemodynamic variables returned to normal within 2 min of releasing the caval cuff. 5. Haematocrit fell during haemorrhage, and remained low for at least 5 days after replacement of the shed blood. Haematocrit was unaffected by simulated haemorrhage. 6. Venous pressure below the inflatable cuff rose by 6 mmHg in the course of simulated haemorrhage. 7. We conclude that the central haemodynamic effects of haemorrhage can be closely and repeatedly simulated by inflating a cuff on the inferior vena cava. This provides a useful technique for repeatedly studying the effects of acute reduction of central blood volume in conscious animals.

Sympathoadrenal mechanisms in cardiovascular responses to naloxone after hemorrhage.

Five rabbits were allotted to each of six treatments on a matched-individual basis. Treatments were none, sham, total adrenalectomy with adrenocorticoid replacement, intravenous guanethidine (15 mg X kg-1 X day-1), adrenalectomy + guanethidine, and adrenal medullectomy. The conscious rabbits were bled 20 ml/kg over 5 min. Naloxone (6 mg/kg) was injected intravenously. The responses of arterial pressure and of plasma epinephrine (E) and norepinephrine (NE) concentrations were measured. Factorial analysis was used to calculate the effects of sympathetic noradrenergic nerves (SYM) and the adrenal medulla (ADR) on the responses. In combination, SYM + ADR fully accounted for the pressor response observed in normal and sham-treated rabbits. SYM and ADR each made independent and approximately equal contributions to the response, but the SYM X ADR interaction was strongly antagonistic. The responses of plasma E and NE were fully accounted for by the adrenal glands and sympathetic nerves, respectively. The pressor responses after total adrenalectomy and adrenal medullectomy were similar. Thus hemorrhage-stimulated adrenal corticosteroid release was not essential to naloxone's action, and adrenal enkephalins were not responsible for naloxone's action on sympathetic pathways.

Factors influencing the effects of intravenous naloxone on arterial pressure and heart rate after haemorrhage in conscious rabbits.

The circulatory responses to different intravenous doses of naloxone were studied in conscious rabbits before and after haemorrhage, under different conditions including prior ganglion blockade. Unless there had been blood loss, naloxone elicited no pressor response, even in high dose. After bleeding so that arterial pressure fell to 40 mmHg, the dose-response relationship for naloxone had two components. Over a low-dose range (threshold 0.3 mg/kg) naloxone had a modest pressor effect but did not affect heart rate. Over a much higher dose range (threshold 0.6 mg/kg) naloxone caused a marked rise in arterial pressure and a profound bradycardia. The highest dose of naloxone examined (25 mg/kg) caused a rise in arterial pressure of 70 mmHg and a reduction in heart rate of 160 beats/min. The pressor and bradycardic effects of naloxone were the same whether post-haemorrhagic hypotension lasted 5, 10, 20 or 30 min. The responses to naloxone in low or high dose depended much more closely on the volume of blood removed than on the level to which arterial pressure fell. Even after non-hypotensive haemorrhage a high dose of naloxone had marked pressor and bradycardic effects. Ganglion blockade prior to haemorrhage abolished the pressor response to a low, but not to a high, dose of naloxone. It was concluded that prolonged and severe hypotension are not necessary to 'prime' the cardiovascular system to respond to naloxone after haemorrhage. In a high dose its pressor effects appear to be mediated post-ganglionically, but in a low dose it may act within the central nervous system.

Circulatory changes during spontaneous motor activity: role of arterial baroreflexes.

Systemic arterial pressure (SAP) and heart rate (HR) were continuously measured in rabbits during spontaneous motor activity such as postural change, exploration, grooming, and eating, and during interposed periods of inactivity within which SAP and HR were defined as basal. Six rabbits were observed for 2 h under each of 3 conditions of arterial baroreceptor afferents: all intact (B4); one carotid sinus intact (B1); all interrupted (B0). In B4, SAP and HR were above basal levels 82% of the time; in B1, 64% and 76% of the time, respectively; in B0, only 21% and 28% of the time, respectively. Supplementary experiments in another six rabbits established that the activity-associated falls of SAP in condition B0 were independent of HR, were not due to engagement of cardiac receptor reflexes, and were not abolished by autonomic ganglion blockade. We conclude that the transient rises of SAP and HR that occur during everyday activity are associated with partial or complete suppression of the reflex effects of arterial baroreceptor input, and we suggest that this is due to transient upward resetting of the arterial baroreflex.

Effects of acute versus chronic deletion of arterial baroreceptor input on the cardiovascular responses to exercise in the rabbit.

These experiments were designed to confirm that at the onset of treadmill exercise in rabbits the tonic reflex depressor effects of input to the central nervous system from arterial baroreceptors is abolished, thus contributing to the rise of systemic arterial pressure (SAP) and heart rate (HR). An inflatable cuff was placed around one common carotid artery after the remaining arterial baroreceptors had been surgically denervated. Transient inflation of the cuff caused reflex rises of SAP and HR, which were much reduced during the first minute of exercise. Deflation of the cuff caused a brisk fall of HR, which was completely abolished by exercise. A snare was placed around one carotid sinus nerve after the remaining arterial baroreceptors had been surgically denervated. Where the snare was tightened all arterial baroreceptor reflexes were immediately and permanently abolished. This allowed the reflex effects of baroreceptor input to be calculated by difference. The magnitude of these calculated effects, at rest and during exercise, diminished according to how long after barodenervation the observations were made. We conclude from the above experiments that the resting tonic reflex depression of SAP and HR caused by baroreceptor input is much reduced, rather than completely abolished, at the onset of exercise. We also conclude, from the effects of partial surgical barodenervation, and of unloading the carotid baroreceptors prior to exercise by inflating the carotid cuff, that resting input from the arterial baroreceptors must be near zero before the cardiovascular response to exercise is grossly altered.

Central suppression of the arterial baroreflexes during spontaneous behaviour.

When input to the central nervous system from arterial baroreceptors was intact, or reduced to one carotid sinus, spontaneous behavioural activities in rabbits caused transient rises of systemic arterial pressure (SAP) and heart rate (HR) from low, basal levels. Complete elimination of baroreceptor input caused an immediate reversal of this pattern, so that the same activities caused abrupt falls of SAP and HR from high basal levels. This same reversal of the SAP response occurred if basal SAP was kept low by sodium nitroprusside, if cardiac nerves were blocked, or if HR was rendered invariable by propranolol and methscopolamine. It is concluded that activity-associated rises of SAP and HR can be accounted for by central suppression of baroreflex mechanisms.

Acute deletion of arterial baroreceptor input in the conscious rabbit.

We describe the use of a snare to denervate the carotid sinus in conscious rabbits whose other arterial baroreceptors had been denervated 8-14 days previously by surgical operation. Within 2 min of tightening the snare arterial blood pressure had risen from 81 +/- 2 to 119 +/- 5 mmHg, and heart rate had risen from 214 +/- 9 to 280 +/- 9 beats min-1. They remained at these high levels for 15 min but then fell, so that after 30 min blood pressure and heart rate were 103 +/- 9 mmHg and 265 +/- 8 beats min-1 respectively. They declined somewhat further, but irregularly, over the succeeding 7 days. The previously brisk baroreceptor-heart rate and carotid sinus reflexes were abolished by ensnaring the carotid sinus nerve, and remained absent over the succeeding 7 days. This method of deleting input from the arterial baroreceptors is not confounded by general anaesthesia or surgical injury, and allows observations to be made within minutes. The rapid decline in blood pressure and heart rate after 15 min, and the subsequent slow downward trend, are unexplained. They are of potential importance in experiments designed to quantitate the effects of baroreceptor input on the circulation. We also noted that, whereas spontaneous activities of the baroreceptor-intact rabbit were usually associated with transient rises of blood pressure and heart rate, after deletion of baroreceptor input the same activities were usually associated with transient falls.