Testosterone facilitates the baroreceptor control of reflex bradycardia: role of cardiac sympathetic and parasympathetic components.

Reported clinical and experimental findings have shown that baroreflex control of heart rate is attenuated in women compared with men. This study investigated whether the sexual dimorphism in baroreflex function relates to the ability of the male hormone testosterone to facilitate baroreflex responsiveness. Relative contributions of the vagal and sympathetic autonomic components to testosterone modulation of baroreflex function were also investigated. Baroreflex curves relating changes in heart rate to increases or decreases in blood pressure evoked by phenylephrine and sodium nitroprusside, respectively, were constructed in sham-operated rats and castrated rats with and without testosterone replacement. Slope of the curves was taken as an index of baroreflex sensitivity (BRS PE and BRS NP ). Castration (for 10 days) significantly reduced plasma testosterone levels and attenuated reflex bradycardia, as indicated by significantly smaller BRS PE in castrated rats compared with values in sham-operated rats (-0.85 +/- 0.07 vs. -1.51 +/- 0.10 beats/min per mm Hg). Testosterone replacement in castrated rats restored plasma testosterone and BRS PE to levels similar to those of sham-operated rats. Muscarinic blockade by atropine caused 55% reduction in BRS PE in sham-operated rats, an effect that was significantly (p < 0.05) attenuated in castrated rats and restored to intact levels after testosterone supplementation. beta-Adrenergic blockade by propranolol caused slight and insignificant decreases in BRS PE. Castration and testosterone supplementation had no effect on BRS NP, ruling out a modulatory effect of testosterone on reflex tachycardia. These data provide the first experimental evidence of a favorable role for testosterone in baroreceptor control of reflex bradycardia. Further, baroreflex modulation by testosterone appears to be autonomically mediated and involves an enhancement of cardiomotor vagal activity.

Evidence for the involvement of central I1 imidazoline receptor in ethanol counteraction of clonidine hypotension in spontaneously hypertensive rats.

Our previous studies have shown that ethanol counteracts centrally mediated hypotensive responses to clonidine. In this study, we investigated the relative roles of central alpha2-adrenergic and I1 imidazoline receptors in the antagonistic ethanol-clonidine hemodynamic interaction. The effects of selective blockade of alpha2- or I1 receptor by 2-methoxyidazoxan and efaroxan, respectively, on the blood pressure and heart rate responses to clonidine and subsequent ethanol administration were evaluated in conscious spontaneously hypertensive rats. Intracisternal administration of clonidine (1.5 microg/kg) produced significant (30 mm Hg; p < 0.05) and sustained (at least 60 min) decreases in blood pressure and heart rate. Systemic ethanol (1 g/kg), administered 10 min after clonidine, counteracted the hypotensive response and restored blood pressure to the preclonidine levels. Treatment with 2-methoxyidazoxan (0.16 microg/kg, intracisternal) or efaroxan (0.45 microg/kg, intracisternal) produced similar attenuation of the hypotensive and bradycardic responses to clonidine. The ability of ethanol to counteract the hypotensive action of clonidine was significantly (p < 0.05) attenuated in rats pretreated with efaroxan. The pressor response to ethanol lasted only 10 min compared with at least 60 min in the absence of efaroxan. In contrast, ethanol counteraction of clonidine-evoked hypotension was not altered when alpha2-adrenoceptors were blocked by 2-methoxyidazoxan. These findings suggest that centrally mediated hypotensive and bradycardic effects of clonidine in conscious spontaneously hypertensive rats involve activation of both alpha2-adrenergic and I1 imidazoline receptors. Furthermore, the findings suggest the dependence of a fully expressed ethanol counteraction of the hypotensive action of clonidine on functional I1 receptor within the central nervous system.

An association between the estrogen-dependent hypotensive effect of ethanol and an elevated brainstem c-jun mRNA in female rats.

We have recently demonstrated that chronic ethanol administration lowers blood pressure (BP) in female rats and this effect is significantly attenuated by ovariectomy. The present study investigated whether ethanol hypotension is estrogen dependent. Further, since estrogen regulates AP-1 activity, the study was extended to determine whether estrogen/c-jun interaction is involved in the estrogen-dependent hypotensive effect of ethanol. Changes in BP and heart rate (HR) were evaluated in radiotelemetered pair-fed sham-operated (SO), ovariectomized (OVX), and OVX estradiol (E2)-treated rats receiving liquid diet with or without ethanol (5%, w/v) for 12 weeks. The in situ hybridization technique was used to measure the c-jun mRNA expression in two brainstem areas, the nucleus tractus solitarius (NTS) and the rostral ventrolateral medulla (RVLM). Ethanol feeding caused significant (P<0.05) decreases in BP in SO rats that started at week 1 and reached its maximum (approximately 10 mmHg) at week 6 and remained at that level till the end of week 12. In OVX rats, ethanol had no effect on BP during the first 5 weeks after which a decrease of 5 mmHg was demonstrated and remained thereafter. Estrogen replacement (17beta-estradiol subcutaneous pellet, 14.2 microg/day) restored the hypotensive effect of ethanol to a level similar to that of SO rats both in terms of magnitude and duration. Densitometric analysis of the in situ hybridization autoradiograms revealed that OVX and E2 replacement had no effect on c-jun mRNA expression in the NTS or RVLM. Ethanol feeding produced a significant (twofold) increase in c-jun mRNA expression in the RVLM of SO rats versus no effect in the NTS. The increased expression of c-jun mRNA observed following ethanol treatment in the RVLM of SO rats was abolished in OVX rats and restored to SO levels after E2 replacement. These findings suggest a link between the estrogen-dependent hypotensive effect of chronically administered ethanol and the increased expression of c-jun mRNA in the brainstem of female rats.

Effect of long-term ethanol feeding on brainstem alpha(2)-receptor binding in Wistar-Kyoto and spontaneously hypertensive rats.

Our previous studies have shown that ethanol attenuates baroreflex function in Wistar-Kyoto (WKY) but not in spontaneously hypertensive rats (SHRs). The present study determined the effects of chronic ethanol administration on alpha(2)-binding sites in brainstem areas that modulate baroreflexes. In vitro autoradiography was utilized to evaluate the effect of a 3-month ethanol feeding on the density (B(max)) and affinity (K(D)) of alpha(2)-adrenoceptors in the middle (mNTS) and rostral (rNTS) portions of the nucleus tractus solitarius of SHRs and WKY rats. Autoradiographic examination of brainstem sections preincubated with [125I]p-iodoclonidine revealed no inter-strain differences in alpha(2)-binding in control rats. Ethanol feeding caused strain-dependent changes in alpha(2)-binding activity, which comprised significant (P<0.05) decreases in the density of alpha(2)-binding sites in both areas of the NTS in SHRs versus no effect in WKY rats. These findings do not favor a role for brainstem alpha(2)-adrenoceptors in ethanol-induced attenuation of baroreflexes. Interestingly, the ethanol-evoked reduction in the NTS alpha(2)-receptor density in SHRs may explain reported findings that ethanol abolishes the hypotensive effect of the alpha(2)-adrenoceptor agonist clonidine in this rat model.

Imidazoline I(1) receptor-induced activation of phosphatidylcholine-specific phospholipase C elicits mitogen-activated protein kinase phosphorylation in PC12 cells.

In the present study, we tested the hypothesis that the activation of imidazoline I(1)-receptor, which is coupled to phosphatidylcholine-specific phospholipase C, results in downstream activation of mitogen-activated protein kinase (p42(mapk) and p44(mapk) isoforms) in PC12 cells. PC12 cells pretreated with nerve growth factor (50 ng/ml, 48 h) to initiate neuronal differentiation were incubated with [methyl-3H]choline and [3H]myristate. Activation of imidazoline I(1) receptor by rilmenidine (10 microM) caused time-dependent increases in diacylglycerol accumulation and phosphocholine release. The Western blotting analysis showed that rilmenidine (10 microM) produced a time-dependent activation of p42(mapk) and p44(mapk) that reached its maximum at 15 min and returned to control levels after 30 min. This finding was confirmed by immunofluorescence labeling of activated mitogen-activated protein kinase in the same model system. Efaroxan (imidazoline I(1)-receptor antagonist) or tricyclodecan-9-yl-xanthogenate (D609, phosphatidylcholine-specific phospholipase C inhibitor) attenuated the phosphorylation of p42(mapk) and p44(mapk) induced by rilmenidine. Nerve growth factor-induced phosphorylation of both mitogen-activated protein kinase isoforms was not affected by D609. These results support the hypothesis that the activation of the imidazoline I(1) receptor coupled phosphatidylcholine-specific phospholipase C results in the downstream activation of mitogen-activated protein kinase.

Clonidine diminishes c-jun gene expression in the cardiovascular sensitive areas of the rat brainstem.

The present study investigated the effect of clonidine on the basal and inducible c-jun and c-fos mRNA expression in the nucleus tractus solitarius (middle, mNTS, and rostral, rNTS) and the rostral ventrolateral medulla (caudal, cRVLM, and rostral, rRVLM). Conscious rats received saline, clonidine (30 microg/kg, i.v.), saline plus sodium nitroprusside (NP), or clonidine plus NP. Under basal conditions (saline-infused rats), c-jun mRNA was expressed in the mNTS and rRVLM but not in the rNTS or cRVLM whereas c-fos mRNA was not detectable. Clonidine attenuated the increases in c-fos in the mNTS and cRVLM and c-jun gene expression in the mNTS and rRVLM caused by NP-evoked hypotension and also reduced the basal expression of c-jun mRNA in the mNTS and rRVLM. These findings establish a causal link between clonidine inhibition of c-fos expression in brainstem and its hypotensive action, and provide the first evidence that clonidine attenuates the expression of the closely linked c-jun gene in neurons implicated in centrally mediated hypotension.

Ovariectomy alters the chronic hemodynamic and sympathetic effects of ethanol in radiotelemetered female rats.

This study determined the chronic hemodynamic effects of ethanol in telemetered freely moving female Sprague-Dawley rats. The role of ovarian hormones and sympathetic activity in the modulation of ethanol responses was also investigated. Changes in blood pressure (BP), heart rate (HR), and plasma estrogen and norepinephrine (NE, as index of sympathetic activity) were evaluated in pair-fed sham-operated (SO) and ovariectomized (OVX) rats receiving liquid diet with or without ethanol (5%, w/v) for 12 weeks. OVX caused a significant increase (about 40 g) in body weight, compared with the sham operation, which was apparent after two weeks and remained so for the duration of the study. The body weight showed gradual and similar increases in both ethanol and control groups. Ethanol feeding had no effect on the plasma estrogen level in SO or OVX rats. Daily ethanol intake was significantly (P < 0.05) less in OVX compared with SO rats whereas the blood ethanol concentration were similar in the two groups except for a significantly (P < 0.05) higher level in OVX rats at weeks 8, 10, and 11. Ethanol feeding caused significant (P < 0.05) decreases in BP in SO rats that started at week land reached maximal response (approximately 10 mmHg) at week 6 and remained at that level till the end of week 12. In OVX rats, ethanol had no effect on BPduring the first 5 weeks of the study. A slight but significant reduction in BP (about 5 mmHg) by ethanol in OVX rats started to appear at week 6 and remained for the following 5 weeks. The reduced hypotensive effect of ethanol in OVX rats was associated with an increase in the sympathetic activity as indicated by the significant (P < 0.05) increases in plasma NE levels. This sympathoexcitatory effect of ethanol was not demonstrated in SO rats. The HR was not affected by ethanol in the two groups of rats except for significant (P < 0.05) increases at weeks 1 through 3 in SO rats. The present findings suggest that the ovarian hormones modulate, at least partly, the hemodynamic and neurohumoral effects of chronic ethanol feeding in female rats. Ethanol lowers BP in female rats and this effect was delayed and diminished in OVX rats due possibly to the associated increase in sympathetic activity.

Radiotelemetric evaluation of hemodynamic effects of long-term ethanol in spontaneously hypertensive and Wistar-Kyoto rats.

This study determined the hemodynamic effects of chronic ethanol in telemetered freely moving age-matched spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats. Changes in blood pressure (BP), heart rate (HR), and plasma norepinephrine (as index of sympathetic activity) were evaluated in pair-fed rats receiving liquid diet with or without ethanol (5%, w/v) for 12 weeks. The SHRs exhibited higher baseline BP and lower HR compared with WKY rats. When normalized for body weight, daily ethanol intake was higher in SHRs compared with WKY rats. However, blood ethanol concentration was similar except for a higher level in SHRs at weeks 7 through 9. Ethanol had no effect on BP in WKY rats but caused decreases in BP in SHRs that reached a maximum (approximately 30 mm Hg) at week 5 and remained thereafter. Ethanol also caused reductions in the BP variability and the circadian fluctuations in BP in SHRs but not in WKY rats. Plasma norepinephrine levels were elevated by ethanol in WKY rats, but not in SHRs. The HR was not affected by ethanol in SHRs and showed increases in WKY rats. These findings suggest that chronic ethanol feeding differentially affects BP in SHRs (hypotension) and WKY rats (no effect). The lack of a hypotensive response to ethanol in WKY rats may relate, at least partly, to the associated sympathoexcitation. The present study used the telemetry technique for BP measurement, which eliminates the confounding and stressful effects of other conventional techniques.

Sexually dimorphic hemodynamic effects of intragastric ethanol in conscious rats.

This study investigated the gender-related differences in hemodynamic effects of small to moderate doses of intragastrically (i.g.) administered ethanol in conscious rats. Changes evoked by ethanol (0.25, 0.5 or 1 g/kg) in mean arterial pressure (MAP), heart rate, cardiac index (CI), stroke volume (SV), and total peripheral resistance (TPR) were followed for 90 min in age-matched male and female Sprague-Dawley rats. Baseline values of MAP (121+/-2 vs. 124+/-2 mm Hg) were similar whereas CI (55+/-2 vs. 43+/-2 ml/min/100 g) and TPR (2.2+/-0.1 vs. 3.0+/-0.1 mm Hg/ml/min/100 g) were significantly (P<0.05) higher and lower, respectively, in female compared with male rats. In male rats, the middle dose (0.5 g/kg) of ethanol caused a slight increase in MAP due to significant (P<0.05) increases in CO whereas the other two doses (0.25 and 1 g/kg) had no effect on MAP. In female rats, MAP was not affected by ethanol (0.25 and 0.5 g/kg) and showed a significant reduction by the higher dose (1 g/kg) that was associated with decreases in CO and SV while TPR did not change. The hypotensive effect of ethanol (1 g/kg) in female rats started after 50 min, was maximal (13+/-1.7 mm Hg) at 70 min and remained so for the remaining 20 min of the study. Blood ethanol concentrations were similar in male and female rats. These findings suggest that the hemodynamic responses to i.g. ethanol are gender-related and that ethanol-evoked hypotension in female rats appears to involve a reduction in cardiac output.

Role of the sympathetic control of vascular resistance in ethanol-clonidine hemodynamic interaction in SHRs.

Our previous studies showed that ethanol selectively counteracts centrally mediated hypotensive responses. In this study, we investigated the role of sympathetic nerve activity, cardiac output (CO), and total peripheral resistance (TPR) in this antagonistic hemodynamic interaction between ethanol and clonidine. Changes in blood pressure (BP), heart rate (HR), CO, stroke volume (SV), and TPR elicited by intracisternal (i.c.) clonidine and subsequent ethanol or saline were evaluated in conscious freely moving spontaneously hypertensive rats (SHRs). Clonidine (0.5 microg, i.c.) evoked hypotension was due to a significant reduction in TPR (from 3.6+/-0.21 to 2.8+/-0.17 mm Hg/ml/min/100 g), which was associated with a significant (p < 0.05) reduction in plasma norepinephrine (NE, from 660+/-115 to 310+/-50 pg/ml), measured as index of sympathetic activity. Ethanol (1 g/kg, i.v.) counteracted the hypotensive effect of clonidine and produced significant (p < 0.05) increases in plasma NE and TPR. Further support for the hypothesis that ethanol selectively counteracts centrally mediated hypotension was sought by investigating the effect of ethanol on peripherally mediated hemodynamic responses to hydralazine. Hydralazine (0.4 mg/kg, i.v.) produced a hypotension similar in magnitude to that produced by clonidine, which was also due to a significant reduction in TPR. However, unlike the case with clonidine, reflex increases in HR, SV, and hence CO were evident. Ethanol given after hydralazine produced a short-lived pressor effect (<10 min vs. 60 min in case of clonidine) in spite of a sustained increase in TPR. The latter was offset by the simultaneous decreases in CO, SV, and HR. A 30% increase in plasma NE caused by hydralazine returned to baseline level after ethanol or saline. Blood ethanol concentrations were similar in all treatment groups. These findings suggest that ethanol selectively counteracts centrally evoked hypotensive responses by counteracting the sympathoinhibition-mediated decreases in TPR elicited by centrally administered clonidine in conscious SHRs.

Estrogen-dependent hypotensive effects of ethanol in conscious female rats.

This study investigated the role of estrogen in the acute hemodynamic responses to intragastric (i.g.) ethanol in conscious female rats. Changes evoked by ethanol or equal volume of water in mean arterial pressure, heart rate, cardiac index, stroke volume (SV), and total peripheral resistance were followed in sham-operated, ovariectomy (OVX) vehicle-treated (OVX-veh), and OVX 17beta-estradiol (E2)-treated (OVX-E2) Sprague-Dawley rats. Plasma norepinephrine (NE) was measured as an index of sympathetic activity. In sham-operated rats, ethanol caused significant decreases in mean arterial pressure that were associated with significant reductions in cardiac index and SV, whereas total peripheral resistance was not changed. Measured plasma NE levels were not affected by ethanol except for a significant reduction observed one time. OVX abolished the hypotensive effect of ethanol and the associated decreases in cardiac output, SV, and plasma NE. Treatment of OVX rats with E2 restored the hypotensive and sympathoinhibitory (decreases in plasma NE) responses to ethanol. Blood ethanol concentrations were not affected by OVX or subsequent E2 administration. These findings suggest that intragastric ethanol elicits estrogen-dependent decreases in blood pressure in female rats, which results mainly from a reduction in cardiac output. The mechanism by which ethanol elicits E2-dependent hypotension remains to be determined.

Estrogen enhancement of baroreflex sensitivity is centrally mediated.

We have recently shown that estrogen enhances baroreceptor control of reflex bradycardia in conscious rats. The present study replicated this finding in pentobarbital sodium-anesthetized rats, and the study was extended to investigate whether this effect of estrogen is centrally or peripherally mediated. Hemodynamic responses to electrical stimulation of the central end of the aortic depressor or the vagal efferent nerve were evaluated in pentobarbital sodium-anesthetized sham-operated (SO), ovariectomized (OVX), and OVX estradiol-treated Sprague-Dawley rats. Phenylephrine (1-16 microgram/kg iv) elicited dose-dependent pressor and bradycardic responses. Regression analysis of the baroreflex curves, relating changes in mean arterial pressure and heart rate, revealed a significantly smaller baroreflex sensitivity in OVX compared with SO anesthetized rats (-0.54 +/- 0.05 and -0.91 +/- 0.12 beats. min-1. mmHg-1, respectively; P < 0.05). Treatment of OVX rats with 17beta-estradiol (E2, 50 microgram. kg-1. day-1 for 2 days subcutaneously) significantly enhanced baroreflex sensitivity to a level similar to that of SO rats (P < 0.05). The enhancing effect of E2 on the baroreflex-mediated bradycardia, observed in conscious and anesthetized rats, seems to be selective because the baroreflex-mediated tachycardic responses measured in a separate group of conscious rats were not altered by ovariectomy or E2 administration. Electrical stimulation of the aortic nerve elicited frequency-dependent depressor and bradycardic responses that were significantly smaller in OVX compared with SO values (P < 0.05). Treatment of OVX rats with E2 restored the hemodynamic responses to aortic stimulation to near SO levels. On the other hand, hemodynamic responses to vagal stimulation were not affected by OVX or treatment with E2. These findings suggest that enhancement of reflex bradycardia by estrogen is centrally mediated and involves interaction with central projections of the aortic nerve.

Ethanol abolishes clonidine-induced impairment of baroreflex control of heart rate in conscious rats.

Our previous studies showed that the ability of ethanol or clonidine to alter the baroreflex control of heart rate (baroreflex sensitivity, BRS) depends on the functional activity of aortic baroreflexes. In this study, we investigated the interaction between the two drugs on BRS in conscious rats with intact baroreflexes (shamoperated, SO) and after aortic baroreceptor denervation (ABD). The slope of the curve relating increments in mean arterial pressure induced by phenylephrine to corresponding reflex bradycardic responses was taken as an index of BRS. Ethanol (1 g/kg i.v.) significantly (p < 0.05) attenuated BRS in SO rats (-1.7 +/- 0.13 versus -1.04 +/- 0.15 beats/min/mm Hg) but not in ABD rats. Clonidine (30 microg/kg, i.v.) elicited significantly (p < 0.05) greater hypotensive responses in conscious ABD compared with SO rats. The BRS was not affected by clonidine administration in SO rats but showed significant (p < 0.05) reductions in ABD rats. Ethanol (1 g/kg, i.v.) had no effect on the hypotensive response to subsequently administered clonidine in ABD and SO rats; however, the effect of the two drugs on BRS was variable. In ABD rats, the BRS values before and after administration of ethanol and clonidine were similar, suggesting that pretreatment with ethanol counteracted clonidine-evoked attenuation of BRS in this rat preparation. In SO rats, the ethanol-clonidine combination produced a significant (p < 0.05) decrease in BRS, similar to the effect of ethanol when administered alone. These data confirm earlier findings that the aortic baroreflex arc modulates the interaction of ethanol and clonidine with baroreflex function. Further, the ability of ethanol to abolish clonidine-induced attenuation of BRS in ABD rats may relate to the compound effects of the two drugs on neuronal pathways participating in the central processing of baroreflexes in these rats.

Acute hemodynamic effects of ethanol in conscious spontaneously hypertensive and normotensive rats.

This study investigated the differential hemodynamic effects of small to high doses of ethanol in conscious age-matched spontaneously hypertensive rats (SHRs) and Wistar Kyoto rats (WKYs). Changes evoked by ethanol (0.25, 0.5, or 1 g/kg, i.v.) or equal volume of saline in mean arterial pressure (MAP), heart rate (HR), cardiac output (CO), stroke volume (SV), and total peripheral resistance (TPR) were followed for 90 min in the two rat strains. The baseline MAP (163 +/- 4 vs. 113 +/- 2 mm Hg) of SHRs was significantly (p < 0.05) higher, compared with WKYs due mainly to the presence of an elevated TPR 13.82 +/- 0.12 vs. 2.51 +/- 0.09 mm Hg/ml/min/100 g, p < 0.05) in SHRs. In both rat strains, all doses of ethanol produced immediate increases in MAP at 1 min, after which the MAP responses varied and depended on the rat strain and dose of ethanol used. In WKYs, 0.25 g/kg ethanol had no effect on MAP, but caused significant decreases in CO and SV and increased HR. Ethanol (0.5 and 1 g/kg) produced a short-lived (10 min) and dose-related increase in MAP. The higher dose (1 g/kg) of ethanol elicited significant (p < 0.05) increases in TPR that were counterbalanced by concomitant decreases in CO and SV. In SHRs, the two higher doses (0.5 and 1 g/kg) of ethanol elicited significant (p < 0.05) decreases and increases in MAP, respectively, compared with control (saline-treated) values. The pressor response to the 1 g/kg dose of ethanol was associated with an increase in TPR that achieved a statistical significance (p < 0.05) at 50 and 80 min after ethanol administration. HR was significantly (p < 0.05) reduced by the two higher doses of ethanol, whereas SV and CO were not changed. Blood ethanol concentrations measured 10, 30, and 60 min after ethanol administration were similar in SHRs and WKYs. These findings suggest that acute administration of ethanol to conscious rats elicits hemodynamic responses that are strain- and dose-dependent. In contrast to a short-lived and dose-related pressor response in WKYs, ethanol (0.5 and 1 g/kg) elicited opposite and longer lasting effects on MAP (decreases and increases, respectively) in SHRs. In both rat strains, the pressor response to the higher dose of ethanol was associated with an increase in TPR; an effect that was compromised by a concomitant decrease in CO in WKYs but not SHRs.

Dopamine modulates peripheral purinergic neurotransmission through multiple presynaptic receptors: tissue-dependent effects.

This study investigated the identity of presynaptic receptors involved in dopaminergic modulation of purinergic transmission in peripheral tissues including isolated rat vas deferens and urinary bladder. Isometric muscle twitches were established in the two tissues by low frequency electric field-stimulation (0.05 Hz, 1-ms duration, and supramaximal voltage). Exposure to prazosin, 50 nmol l-1 (vas deferens), or atropine, 3 micromol l-1 (urinary bladder), had no effect on the developed twitches. In contrast, desensitisation of P2X-purinoceptors by alpha,beta-methylene ATP (alpha,beta-mATP, 30 micromol l-1) abolished the twitches in both tissues, confirming their purinergic origin. Dopamine (1.8x10(-7) to 4.2x10(-5) mol l-1) reduced the twitch response in a concentration-related manner. Yohimbine (alpha2-adrenoceptor antagonist, 0.3 micromol l-1) significantly (P<0.05) attenuated the inhibitory effects of dopamine and caused an upward shift in the concentration-response curves in the vas deferens and the urinary bladder. On the other hand, a blockade of DA2-dopaminoceptors by domperidone (1 micromol l-1) produced significant (P<0.05) reductions in dopamine responses only in rat vas deferens, with no effect in the urinary bladder. These data suggest that dopamine exerts inhibitory influences on purinergically-mediated muscle twitches in rat vas deferens and urinary bladder. More importantly, the nature of presynaptic receptors (alpha2-adrenergic and/or DA2-dopaminergic) involved in mediating dopamine effects is dependent on the tissue under investigation.

Ethanol counteraction of I1-imidazoline but not alpha-2 adrenergic receptor-mediated reduction in vascular resistance in conscious spontaneously hypertensive rats.

Our recent findings have shown that ethanol selectively counteracts decreases in blood pressure (BP) evoked via activation of central I1-imidazoline receptors but not alpha-2 adrenoceptors in conscious spontaneously hypertensive rats (SHRs). This study investigated the role of sympathetic activity, cardiac output and total peripheral resistance (TPR) in the differential effect of ethanol on centrally mediated hypotension. Changes in plasma norepinephrine (NE), as index of sympathetic activity, BP, heart rate, cardiac index, stroke volume, and TPR elicited by rilmenidine or alpha-methylnorepinephrine (selective I1 and alpha-2 receptor agonists, respectively) and subsequent ethanol (0.5 or 1 g/kg) or saline, were evaluated in conscious SHRs. Intracisternal rilmenidine (25 microg) or alpha-methylnorepinephrine (alpha-MNE; 4 microg) elicited similar decreases in BP, TPR, and plasma NE, but cardiac index was not changed. Ethanol (0.5 g/kg i.v.) had no effect on hemodynamic responses to rilmenidine or alpha-MNE. The higher dose (1 g/kg i.v.) of ethanol counteracted the hypotensive response to rilmenidine and significantly (P <.05) elevated TPR and plasma NE. In contrast, ethanol (1 g/kg) had no effect on the hypotensive responses to alpha-MNE but significantly (P <.05) elevated plasma NE. However, this increase in NE was approximately one third of the increase evoked by ethanol when given after rilmenidine. These findings suggest that the selective counteraction by ethanol of the hypotension evoked via activation of central I1 but not alpha-2 receptors may relate, at least in part, to its greater ability to reverse the sympathoinhibition and the associated decrease in vascular resistance mediated by I1 receptors.

Estrogen enhances baroreflex control of heart rate in conscious ovariectomized rats.

In previous studies, we have shown that the baroreflex control of heart rate is significantly attenuated in females compared with age-matched males. This study investigated the role of estrogen in the modulation of baroreflex function in conscious unrestrained rats. Baroreflex-mediated decreases in heart rate in response to increments in blood pressure evoked by phenylephrine were evaluated in conscious freely moving male and female Sprague-Dawley rats as well as in ovariectomized rats. The effect of a 2-day 17 beta-estradiol (50 micrograms.kg-1.day-1, s.c.) or vehicle treatment on baroreflex sensitivity was investigated in ovariectomized rats. Intravenous bolus doses of phenylephrine (1-16 micrograms/kg) elicited dose-dependent pressor and bradycardic responses in all groups of rats. Regression analysis of the baroreflex curves relating increments in blood pressure to the associated heart rate responses revealed a significantly (p < 0.05) smaller baroreflex sensitivity in female compared with male rats (-1.22 +/- 0.07 and -1.85 +/- 0.15 beats.min-1.mmHg-1, respectively), suggesting an attenuated baroreflex function in females. In age-matched ovariectomized rats, baroreflex sensitivity showed further reduction (-0.93 +/- 0.02 beats.min-1.mmHg-1). Treatment of ovariectomized rats with 17 beta-estradiol significantly (p < 0.05) enhanced the baroreflex sensitivity (-1.41 +/- 0.16 beats.min-1.mmHg-1) to a level that was slightly higher than that of sham-operated female rats. Furthermore, baroreflex sensitivity of ovariectomized estradiol-treated rats was not significantly different from that of age-matched male rats. The vehicle, on the other hand, had no effect on baroreflex sensitivity of ovariectomized rats. These data support our earlier findings that sexual dimorphism exists in baroreflex control of heart rate. More importantly, the present study provides experimental evidence that suggests a facilitatory role for estrogen in the modulation of baroreflex function.

Ethanol selectively counteracts hypotension evoked by central I(1)-imidazoline but not alpha2-adrenergic receptor activation in spontaneously hypertensive rats.

Previous studies from our laboratory showed that ethanol counteracts hypotensive responses to clonidine in spontaneously hypertensive rats. This study investigated whether this effect of ethanol involves interaction with central alpha2-adrenoceptors or I(1)-imidazoline receptors or both. The effects of ethanol (0.5 or 1 g/kg, i.v.) or an equal volume of saline on hypotensive and bradycardic responses to clonidine (mixed alpha2-adrenoceptor/I(1)-imidazoline receptor agonist), rilmenidine (selective I(1)-imidazoline receptor agonist), or alpha-methylnorepinephrine (selective alpha2-adrenoceptor agonist) were studied in conscious spontaneously hypertensive rats. Intracisternal administration of clonidine (0.5 microg), rilmenidine (25 microg), or alpha-methylnorepinephrine (4 microg) elicited similar decreases in mean arterial pressure (MAP; 25-30 mm Hg) that lasted > or =60 min. Subsequent administration of ethanol (0.5 and 1 g/kg, i.v.) counteracted the hypotensive effect of clonidine in a dose-related manner. Ethanol (1 g/kg) increased the blood pressure to levels similar to baseline (preclonidine) levels, and blood pressure remained significantly (p < 0.05) higher compared with the corresponding values in saline-treated rats. Similarly, ethanol (0.5 and 1 g/kg, i.v.) dose-dependently counteracted the hypotensive effect of rilmenidine. The antagonizing effects of ethanol on hypotension evoked by clonidine and rilmenidine were comparable in terms of both magnitude and duration. In contrast, ethanol (0.5 or 1 g/kg) had no effect on hypotension evoked by alpha-methylnorepinephrine. Except for a brief increase in blood pressure by ethanol (1 g/kg) at 5 min, blood pressure values obtained in alpha-methylnorepinephrine-treated rats receiving any of the two doses of ethanol were similar to postsaline values. Ethanol had no effect on bradycardic responses to any of the three hypotensive agents. Blood ethanol concentrations were similar regardless of the antihypertensive drug used. We concluded that the adverse hemodynamic effect of ethanol on centrally mediated hypotensive responses depends on the types of receptors involved in the elicitation of this response. That ethanol counteracts decreases in blood pressure evoked by clonidine and rilmenidine but not by alpha-methylnorepinephrine suggests an interaction between ethanol and central pathways involved in I(1)-imidazoline receptor-mediated hypotension.

Ovariectomy abolishes ethanol-induced impairment of baroreflex control of heart rate in conscious rats.

Our previous studies have shown that ethanol attenuates baroreflex control of heart rate in male rats. The present study investigated whether this effect of ethanol is gender-related, and whether it involves hormonal factors. The effect of intragastric administration of ethanol or equal volume of water on baroreflex-mediated decreases in heart rate in response to increments in blood pressure evoked by phenylephrine were evaluated in conscious age-matched male and female Sprague-Dawley rats as well as in ovariectomized rats. Baroreflex curves relating changes in blood pressure and associated heart rate responses were constructed, and the slopes of the regression lines were taken as a measure of baroreflex sensitivity. Phenylephrine (1-16 microg kg(-1), i.v.) elicited dose-dependent pressor responses that were similar in all groups of rats. However, the associated reflex bradycardic responses depended on the rat preparation and the dose of ethanol employed. In water-treated (control) animals, significantly (P < 0.05) lesser reflex bradycardic responses were observed in female compared with male rats (baroreflex sensitivity, -1.21 +/- 0.12 vs. -1.67 +/- 0.12 beats min(-1) mmHg(-1)). Ovariectomy resulted in a further reduction in baroreflex sensitivity (-0.82 +/- 0.06 beats min(-1) mmHg(-1)), suggesting a favorable role for ovarian hormones in baroreflex modulation. In male rats, ethanol (0.25, 0.5, or 1 g kg(-1), intragastric) elicited dose-related decreases in reflex bradycardic responses. The reduction in the regression coefficient obtained by the two higher doses (0.5 and 1 g kg(-1)) of ethanol was statistically significant compared with control values. The ability of ethanol to reduce baroreflex sensitivity appears to be gender-independent as it was similarly demonstrated in intact female rats. In contrast, ethanol had no effect on reflex bradycardic responses in ovariectomized rats at any of the doses tested. The data suggest that ethanol reduces baroreflex control of heart rate irrespective of the rat gender. Further, the lack of an effect of ethanol on baroreflex sensitivity in ovariectomized rats may suggest a role for ovarian hormones in ethanol-evoked baroreflex attenuation.

Facilitation of reflex bradycardia does not contribute to the enhanced hypotensive effect of clonidine in aortic barodenervated rats.

Our previous studies showed that the hypotensive effect of clonidine is enhanced in rats with surgically eliminated aortic baroafferents. In this study, we investigated whether this effect of clonidine is related to facilitation of baroreceptor control of reflex bradycardia. The effects of clonidine on blood pressure (BP), heart rate (HR), and baroreflex-mediated decreases in HR (baroreflex sensitivity, BRS) were studied in conscious aortic barodenervated (ABD) and sham-operated (SO) rats. The slope of the baroreflex curve relating increments in mean arterial pressure (MAP) induced by phenylephrine to corresponding baroreflex-mediated bradycardic responses was taken as an index of BRS. ABD but not the sham operation caused immediate and significant (p < 0.05) increases in BP and HR and an impairment of BRS. Two days after ABD, these parameters, except the BRS, subsided to near control levels. Starting from similar baseline values of BP and HR, clonidine (30 microg/kg, i.v.) elicited significantly (p < 0.05) greater decreases in MAP in conscious ABD rats compared with SO rats (-23 +/- 2 mm Hg vs. -7 +/- 2 mm Hg). The enhanced hypotensive effect of clonidine in ABD rats was associated with a significant reduction in baroreceptor-mediated reflex bradycardic responses to increments in BP evoked by phenylephrine. The slope of the baroreflex curves that represented the BRS showed approximately 40% reduction after treatment with clonidine (baseline BRS, -1.2 +/- 0.06 beats/min/mm Hg; clonidine, -0.73 +/- 0.07 beats/min/mm Hg). On the other hand, a threefold lower decrease in BP by clonidine in SO rats was not associated with any alteration in BRS. These findings support the hypothesis that aortic baroreceptors exert a tonically active restraining influence on centrally mediated hypotension. More important, the results do not favor a role for facilitation of baroreflexes in the enhanced hypotensive effect of clonidine in denervated rats.