Norepinephrine transporter-deficient mice respond to anxiety producing and fearful environments with bradycardia and hypotension.

The study of anxiety and fear involves complex interrelationships between psychiatry and the autonomic nervous system. Altered noradrenergic signaling is linked to certain types of depression and anxiety disorders, and treatment often includes specific transporter blockade. The norepinephrine transporter is crucial in limiting catecholaminergic signaling. Norepinephrine transporter-deficient mice have increased circulating catecholamines and elevated heart rate and blood pressure. We hypothesized, therefore, that reduced norepinephrine clearance would heighten the autonomic cardiovascular response to anxiety and fear. In separate experiments, norepinephrine transporter-deficient (norepinephrine transporter-/-) mice underwent tactile startle and trace fear conditioning to measure hemodynamic responses. A dramatic tachycardia was observed in norepinephrine transporter-/- mice compared with controls following both airpuff or footshock stimuli, and pressure changes were also greater. Interestingly, in contrast to normally elevated home cage levels in norepinephrine transporter-deficient mice, prestimulus heart rate and blood pressure were actually higher in norepinephrine transporter+/+ animals throughout behavioral testing. Upon placement in the behavioral chamber, norepinephrine transporter-deficient mice demonstrated a notable bradycardia and depressor effect that was more pronounced in females. Power spectral analysis indicated an increase in low frequency oscillations of heart rate variability; in mice, suggesting increased parasympathetic tone. Finally, norepinephrine transporter-/- mice exhibited sexual dimorphism in freeze behavior, which was greatest in females. Therefore, while reduced catecholamine clearance amplifies immediate cardiovascular responses to anxiety- or fear-inducing stimuli in norepinephrine transporter-/- mice, norepinephrine transporter deficiency apparently prevents protracted hemodynamic escalation in a fearful environment. Conceivably, chronic norepinephrine transporter blockade with transporter-specific drugs might attenuate recognition of autonomic and somatic distress signals in individuals with anxiety disorders, possibly lessening their behavioral reactivity, and reducing the cardiovascular risk factors associated with persistent emotional arousal.

Animal model of neuropathic tachycardia syndrome.

Clinically relevant autonomic dysfunction can result from either complete or partial loss of sympathetic outflow to effector organs. Reported animal models of autonomic neuropathy have aimed to achieve complete lesions of sympathetic nerves, but incomplete lesions might be more relevant to certain clinical entities. We hypothesized that loss of sympathetic innervation would result in a predicted decrease in arterial pressure and a compensatory increase in heart rate. Increased heart rate due to loss of sympathetic innervation is seemingly paradoxical, but it provides a mechanistic explanation for clinical autonomic syndromes such as neuropathic postural tachycardia syndrome. Partially dysautonomic animals were generated by selectively lesioning postganglionic sympathetic neurons with 150 mg/kg 6-hydroxydopamine hydrobromide in male Sprague-Dawley rats. Blood pressure and heart rate were monitored using radiotelemetry. Systolic blood pressure decreased within hours postlesion (Delta>20 mm Hg). Within 4 days postlesion, heart rate rose and remained elevated above control levels. The severity of the lesion was determined functionally and pharmacologically by spectral analysis and responsiveness to tyramine. Low-frequency spectral power of systolic blood pressure was reduced postlesion and correlated with the diminished tyramine responsiveness (r=0.9572, P=0.0053). The tachycardia was abolished by treatment with the beta-antagonist propranolol, demonstrating that it was mediated by catecholamines acting on cardiac beta-receptors. Partial lesions of the autonomic nervous system have been hypothesized to underlie many disorders, including neuropathic postural tachycardia syndrome. This animal model may help us better understand the pathophysiology of autonomic dysfunction and lead to development of therapeutic interventions.

Modulatory effects of endothelin on baroreflex activation in the nucleus of the solitary tract.

In this study, we determine the effects of endogenous endothelin on baroreflex activation. After control baroreflex slopes were obtained, the animals received bilateral intra-nucleus tractus solitarii microinjections of saline, or equimolar doses (4 pmol/60 nl) of the endothelin ETA receptor antagonist cyclo (D-Trp-D-Asp-Pro-Val-Leu (BQ-123), Homopiperinidinyl-CO-Leu-D-Trp(CHO)-D-Trp-OH (BQ-610), or the endothelin ETB receptor antagonist N-cis-2,6-dimethylpiperidinocarbonyl-L-gamma-MeLeu-D-Trp( COOCH3)-D-Nle (BQ-788). Intra-nucleus tractus solitarii administration of BQ-123 resulted in a brief initial pressor effect followed by hypotension which resolved by 15 min. The baroreflex slope was significantly enhanced when tested 15 min after BQ-123 treatment (from 2.4 +/- 0.5 ms/mmHg to 3.5 +/- 0.4 ms/mmHg). Similar effects were observed with the other endothelin ETA receptor antagonist, except that the hypertensive and hypotensive responses were more pronounced while the baroreflex slope was similarly increased. In contrast, the endothelin ETB receptor antagonist did not evoke appreciable changes in hemodynamics or in baroreflex slopes. Our results support the concept that endothelin prominently affects reflex cardiovascular function through the endothelin ETA receptor subtype.

Inhibition by ethanol of the cardiovascular effects of glutamate in the nucleus of the solitary tract.

This study investigates whether ethanol affects the cardiovascular changes evoked by the excitatory amino acid glutamate in the nucleus of the solitary tract (NTS). Male Sprague-Dawley rats were anesthetized with urethane and instrumented for microinjection of drugs into the NTS. In 28 animals, an initial dose-response curve for glutamate (37, 74, and 148 pmol/60 nL) was obtained. In eight animals (control group), the dose-response curve was unchanged when it was tested after microinjection of 60 nL of saline into the NTS. In contrast, the prior intra-NTS administration of ethanol (25 or 50 mM) consistently inhibited the hypotensive and bradycardic effects of glutamate. The specificity of this inhibitory response was corroborated by the inability of intra-NTS administrations of ethanol (50 mM) to affect the hypotensive and bradycardic responses to nicotine (922 pmol. 1.84 nmol, and 3.96 nmol). These results indicate that the cardiovascular effects of glutamate can be antagonized by ethanol in the NTS. This inhibitory effect, in turn, may provide a basis for the hypertensinogenic action of ethanol in humans.

Effects of nicotine on brain stem mechanisms of cardiovascular control.

The role of the central nervous system in the pressor effect of nicotine is not well understood. In this study, we evaluated the cardiovascular effects of nicotine in the lower brainstem of normotensive and hypertensive rats. Microinjection of nicotine (0.012-3696 pmol/60 nl) into the nucleus of the solitary tract and area postrema of Sprague-Dawley (SD), Wistar Kyoto (WKY) and spontaneously hypertensive rats (SHR) decreased blood pressure and heart rate. In contrast, administration of similar doses of nicotine within the rostral ventrolateral medulla (RVLM) evoked a long-lasting pressor and tachycardic effect. This pressor effect was completely abolished by prior microinjection of hexamethonium. In SHR the depressor and bradycardic responses in the nucleus of the solitary tract and area postrema were similar to those of normotensive animals. The pressor effect in the RVLM, however, was more pronounced in the SHR than in WKY or SD rats. In additional experiments, the changes produced by intra-RVLM administration of nicotine on renal sympathetic nerve activity, blood pressure and heart rate were evaluated before and after equidepressor intravenous doses of either clonidine, labetalol or prazosin. The prior administration of labetalol antagonized the pressor effect of nicotine in the three strains of rats (SHR, 82 +/- 6%; SD, 96 +/- 4%; WKY, 83 +/- 9%). Prazosin inhibited the nicotine pressor response by 69% in SHR, by 44% in SD and by 70% in WKY. Clonidine had no effect on nicotine response in the three groups of rats. In conclusion, nicotine administration within the RVLM increases renal sympathetic nerve activity and blood pressure.(ABSTRACT TRUNCATED AT 250 WORDS)

Endothelin as a neuropeptide. Cardiovascular effects in the brainstem of normotensive rats.

The relevance of endothelin in central cardiovascular function was studied in urethane-anesthetized Sprague-Dawley rats. Blood pressure (BP) was monitored intra-arterially, and cerebrospinal fluid (CSF) was collected through an intracisternal catheter for radioimmunoassay of endothelin-1 (ET-1). Endothelin levels in the CSF were significantly higher (39 +/- 3 pg/ml) than in plasma (10 +/- 3 pg/ml, n = 11). ET-1 in CSF or plasma was not affected by systemic infusion of saline, but its levels significantly decreased when a sustained increase in BP was elicited with phenylephrine (14 +/- 7 pg/ml in the CSF and 6 +/- 4 pg/ml in plasma, n = 5). In sinoaortic-denervated animals, phenylephrine failed to reduce CSF endothelin levels. In different experiments, intracisternal administration of ET-1 (10 pmol) evoked an initial decrease in BP and heart rate (HR), followed by pronounced hypertension, bradycardia, and, in 70% of the animals, death from cardiorespiratory failure. Intracisternal administration of endothelin-3 (ET-3, 80 pmol, n = 11) evoked only a modest hypotensive and bradycardic response without cardiorespiratory impairment. Microinjection of ET-1 (0.5, 1, 2, 4, and 6 pmol/60 nl) into the nucleus of the solitary tract or area postrema produced a decrease in BP and HR. On the other hand, injection of low concentrations of ET-3 into the nucleus of the solitary tract increased BP and HR (at 2 pmol, 17 +/- 3 mm Hg, 14 +/- 6 beats per minute, n = 7), whereas ET-3 in the area postrema produced a prominent dose-related decrease in BP and HR. In the rostroventrolateral medulla, the lowest doses of ET-1 first modestly increased BP and renal sympathetic nerve activity. These effects were followed by hypotension, bradycardia, increase in respiratory frequency, and further enhancement of sympathetic nerve traffic. In 29% of the animals, these effects were followed by cardiorespiratory arrest. The specificity of the cardiovascular response to endothelin was demonstrated by the inhibitory effects of the receptor antagonist BQ-123. These results demonstrate that endothelin has specific cardiovascular effects in the brainstem of the rat and support a role for endothelin in cardiovascular regulation.

A Na pump inhibitor from bovine posterior pituitary: purification, structure determination and its cardiovascular effect in rat.

We examined the hypothesis that hypothalamo-hypophysial tissue contains an endogenous Na pump inhibitor. From bovine posterior pituitary, we purified a substance which inhibits Rb uptake by human erythrocytes. This inhibitory activity was found in the eluate of 10% acetonitrile from a C18 flash column and purified by subsequent three steps of reversed-phase high-performance liquid chromatography (HPLC). Sequence analysis revealed that this substance was identical to joining peptide, one of the major products of proopiomelanocortin (POMC). This peptide had hypertensive and tachycardiac effects in spontaneously hypertensive rats (SHR) after central administration, with weak Na,K-ATPase inhibitory activity (IC50 = 0.5 mM).

Cardiovascular excitatory effects of adenosine in the nucleus of the solitary tract.

Adenosine is an inhibitory neuromodulator in several brain regions. In the nucleus tractus solitarius (NTS), however, adenosine exerts excitatory cardiovascular effects. The purpose of the present study was to elucidate the involvement of other endogenous mechanisms that could contribute to the final hemodynamic response to adenosine in this nucleus. In normotensive Sprague-Dawley rats, intra-NTS microinjection of adenosine (2.3 nmol/60 nl) decreased blood pressure and heart rate. These effects were blocked by prior administration of the specific adenosine receptor antagonist 1,3-dipropyl-8-p-sulfophenylxanthine (0.92 nmol) and by the two glutamate receptor antagonists kynurenic acid and glutamic diethylester. The specificity of the adenosine-glutamate interaction in the NTS was demonstrated with adrenergic and angiotensin receptor antagonists that did not affect the adenosine response and by experiments with glutamate receptor antagonists that did not affect nicotine actions in the NTS. Furthermore, an increase in glutamate levels was demonstrated during perfusion of adenosine through a microdialysis probe in the NTS of anesthetized rabbits. These findings indicate that adenosine increases the release of glutamate in the NTS and, thus, are at variance with the concept of a "universal" inhibitory effect of adenosine in the central nervous system.

Administration of carbamazepine in the nucleus of the solitary tract inhibits the antihypertensive effect of clonidine.

The effects of carbamazepine administration into the nucleus tractus solitarii (NTS) on central alpha 2-adrenergic cardiovascular function were studied in normotensive Wistar Kyoto (WKY) and spontaneously hypertensive rats (SHR). Stereotaxic microinjections of carbamazepine (288 ng/60 nl) into the NTS of SHR transiently increased blood pressure, heart rate, and renal sympathetic nerve activity. These effects were followed by a modest depressor and bradycardic effect, with no change in sympathetic nerve activity. In addition, previous intra-NTS administration of carbamazepine attenuated the antihypertensive effect of the centrally acting alpha 2-adrenoceptor agonist, clonidine. Similar carbamazepine effects were observed in the WKY rats. These results suggest that some of the cardiovascular changes observed during carbamazepine treatment involve antagonism of central noradrenergic mechanisms.

Cardiovascular effects of microinjection of angiotensin II in the brainstem of renal hypertensive rats.

The cardiovascular effects of microinjection of angiotensin II (AII) into the area postrema (AP), nucleus of the solitary tract (NTS) and rostroventrolateral medulla were studied in urethane anesthetized sham-normotensive (NT) and two-kidney, one-clip renal hypertensive rats. Microinjection of AII (2-2000 ng) in the AP of renal hypertensive rats elicited a dose-dependent decrease in blood pressure, heart rate and renal sympathetic nerve activity. Similar effects were observed in the NTS. In the NT rats, low doses of AII (2 and 20 ng), either in the AP or NTS, were also depressor. High doses of AII (200-2000 ng) were needed to observe a modest pressor effect in the NT animals. A decrease in heart rate and renal sympathetic activity was observed with the pressor effect. The AII-antagonist, [Sar1,Val5,Ala8]-AII, into the NTS or AP increased blood pressure and heart rate and inhibited the cardiovascular effects of low doses of AII in both group of rats. In contrast, [Sar1,Val5,Ala8]AII did not affect the pressor action of high doses of AII in the NT group. While the microinjection of AII into the rostroventrolateral medulla did not produce any significant cardiovascular effect in the renal hypertensive group, it resulted in a modest pressor effect in the NT rats. These results indicate that acute activation of AII receptors in the AP or NTS does not contribute to the pressor effect of AII in renal hypertensive rats.

Modulatory effects of adenosine on baroreflex activation in the brainstem of normotensive rats.

The effects of the adenosine antagonists, 1,3-dipropyl-8-p-sulphenylxanthine (DPSPX) and caffeine, on baroreflex activity were tested in normotensive Sprague-Dawley rats. The microinjection of DPSPX (0.92 nmol) into the nucleus tractus solitarii (NTS) of urethane-anesthetized animals did not modify basal blood pressure or heart rate but inhibited the reflex bradycardia elicited by phenylephrine. Similar inhibitory effects on baroreflex activation were observed after intracisternal administration of caffeine to conscious or anesthetized animals. These results suggest that central endogenous adenosine is involved in the medullary regulation of blood pressure and that adenosine antagonists such as caffeine can inhibit baroreflex activation.

Cardiovascular effects of neuropeptide Y in rat brainstem nuclei.

Central catecholaminergic neurons are involved in cardiovascular regulation. Neuropeptide Y (NPY) coexists with adrenaline and noradrenaline in the rat brain, and interactions among these substances have been studied. The purpose of this study was to investigate the possible role of NPY in central cardiovascular control. Male Sprague-Dawley rats were anesthetized with urethane, and blood pressure was monitored intra-arterially. Intramedullary microinjection (60 nl) of NPY (0, 46.5 fmol, 465 fmol, 1.5 pmol, and 4.65 pmol) was made into the nucleus tractus solitarii (NTS), into the area postrema, and into the C1 area in the rostroventrolateral medulla. Injection site was identified by L-glutamate administration and confirmed histologically. Unilateral injection of NPY into the NTS produced a prominent dose-related decrease in heart rate and systolic and diastolic blood pressure (-106 +/- 8 beats/min, -56 +/- 2 mm Hg, and -33 +/- 2 mm Hg, respectively after 4.65 pmol NPY, n = 7, p less than 0.001). Maximal changes occurred at 30 seconds and recovered in 10 minutes for blood pressure and 20 minutes for heart rate. Injection into the area postrema produced an initial increase in heart rate and mean blood pressure (+23 +/- 2 beats/min and +18 +/- 2 mm Hg) followed by a prolonged decrease in heart rate and mean blood pressure (-14 +/- 4 beats/min and -15 +/- 2 mm Hg, respectively, n = 7, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Purinergic receptors in the brainstem mediate hypotension and bradycardia.

Adenosine acts at many sites to modulate neuronal activity. The purpose of this study was to investigate a possible role for adenosine as a neuromodulator of brainstem cardiovascular control. Microinjections of adenosine (0-2.3 nmol) were made stereotaxically into various brainstem sites. Injection of adenosine into the nucleus tractus solitarii (NTS) produced dose-related decreases in heart rate and systolic and diastolic blood pressures. Maximal changes occurred 90 seconds after injection. Injection into the area postrema also produced decreased heart rate and systolic and diastolic blood pressures. No significant effect occurred following injection into the C1 area. Adenosine 5'-triphosphate and its analogue, beta, gamma-methylene adenosine 5'-triphosphate also produced dose-related and potent vasodepressor and bradycardia effects in the NTS. Injection of 1,3-dipropyl-8-p-sulfophenylxanthine (0.92 nmol), a potent adenosine receptor antagonist, produced no effect itself, but abolished for 45 minutes the actions of further injections of adenosine and adenosine 5'-triphosphate (but not L-glutamate) in both the NTS and area postrema. Thus, NTS and area postrema injections of adenosine decrease blood pressure and heart rate in anesthetized normotensive rats through adenosine receptors located in these areas. These findings support a role for endogenous adenosine as a central modulator in cardiovascular control.

Smoking and mechanisms of cardiovascular control.

In humans short-term administration of nicotine, whether by smoking or intravenous injection, will typically raise blood pressure by 5 to 10 mm Hg and heart rate by 10 to 25 bpm. Smoking causes reduced myocardial contractility and left ventricular function in patients with angina pectoris or heart failure. Nicotine's mechanism of action is more complex than the classic concept of nicotinic ganglionic stimulation can account for. Nicotine exerts a potent pressor effect in the ventral lateral medulla (C-1 area). Little current data are available documenting the efficacy of centrally acting antihypertensive agents and converting-enzyme inhibitors with regard to preventing nicotine's acute cardiovascular effects.