A novel mechanism prevents the development of hypertension during chronic cold stress.

1 Chronic cold exposure of rats (7 days in a cold room at 4 degrees C) attenuated the sympathetic nerve stimulation (NS)-induced overflow of noradrenaline (NE) (measured by high-performance liquid chromatography, coupled to electrochemical detection) appearing in the perfusate/superfusate of the perfused mesenteric arterial bed as well as the increase in the perfusion pressure. 2 The same type of cold exposure resulted in an increase in tyrosine hydroxylase (TH) gene expression measured in the superior cervical ganglion and NE content measured in the mesenteric artery obtained from cold-exposed rats. 3 Addition of sodium nitroprusside, a nitric oxide (NO) donor, to the buffer perfusing the mesenteric arterial bed obtained from rats maintained at room temperature also resulted in an attenuation of the NS-induced overflow of NE and increase in perfusion pressure. 4 N(c)-nitro-L-arginine methyl ester (L-NAME), an NO synthase inhibitor, placed in the drinking water prevented the attenuation of the pre- and post-junctional responses to NS of the mesenteric arterial bed obtained from cold-exposed rats. 5 L-NAME treatment also increased the cold-induced elevation of blood pressure seen in whole animals. 6 The present results are consistent with the idea that cold exposure leads to a concomitant increase in sympathetic nerve activity and production of NO. We hypothesize that the increase in production and release of NO results in a decrease in the biologically active form of NE despite increased synthesis and release of the catecholamine. 7 It is concluded that the above-mentioned interactions serve as a protective mechanism offsetting the increased release and action of NE from sympathetic nerves and thus preventing the development of hypertension.

Role of neuropeptide Y in cold stress-induced hypertension.

Chronic cold stress (4 degrees C) produced a sustained increase in mean arterial pressure in both normotensive and borderline hypertensive rats (BHR). The high blood pressure in BHRs was significantly reversed by a neuropeptide Y (NPY) Y1 receptor antagonist suggesting that NPY is involved in mediating stress-induced hypertension. Corresponding increases in adrenal NPY messenger RNA and NPY immunoreactivity were found during the stress; furthermore, chronic cold stress also potentiated the pressor response of rats to a subsequent acute stress test in which NPY has been shown to play a role. These results suggest that chronic cold stress-induced hypertension is mediated by elevated NPY release and vascular tone as a result of increased NPY gene expression and storage.

Direct evidence for the role of neuropeptide Y in sympathetic nerve stimulation-induced vasoconstriction.

Neuropeptide Y (NPY) is a vasoconstrictor peptide and a cotransmitter with norepinephrine (NE) in sympathetic nerve terminals and is thought to be involved in sympathetic nerve stimulation (SNS)-induced vasoconstriction. Using BIBP-3226, a Y1 receptor selective antagonist, we examined this hypothesis in the isolated and perfused mesenteric vascular bed. SNS produced a frequency-dependent increase in perfusion pressure and concomitant overflow of NPY immunoreactivity in the perfusate. [Leu31,Pro34]NPY potentiated NE-induced and ATP-induced vasoconstriction, indicating the presence and biological action of Y1 receptors in this vascular bed. The potentiation effect of [Leu31,Pro34]NPY of the increase in perfusion pressure by NE, ATP, or SNS was prevented by BIBP-3226. In addition, SNS-induced vasoconstriction at both high and low frequencies was significantly attenuated by BIBP-3226 at a concentration that completely blocked the [Leu31,Pro34]NPY-induced potentiation of the NE- or ATP-induced vasoconstrictor effect. These results suggest that approximately 30% of vasoconstriction produced by SNS depends on NPY in the mesenteric vascular bed.

Elevated neuropeptide Y gene expression and release during hypoglycemic stress.

Our previous studies show that neuropeptide Y (NPY) is involved in mediating sympathetic nerve stimulation-induced vasoconstriction. Insulin hypoglycemia is known to produce increased sympathetic output and elevated arterial pressure. The present study examined the role of NPY in the hypertensive response to insulin by examining the effects of insulin on NPY gene expression, tissue content and release. Subcutaneous injection of insulin produced an immediate increase in plasma NPY immunoreactivity (NPYir) and delayed increases in adrenal and neuronal NPY mRNA and adrenal NPYir in rats. These results suggest that NPY may play a role in insulin-induced hypertension.

Calcitonin gene-related peptide is the endogenous mediator of nonadrenergic-noncholinergic vasodilation in rat mesentery.

In the isolated perfused rat mesenteric vascular bed pretreated with guanethidine and precontracted with methoxamine, periarterial nerve stimulation elicited a frequency-dependent and endothelium-independent vasodilation. The sustained vasodilation was slow-onset and reversible. It was resistant to propranolol or atropine but sensitive to tetrodotoxin and capsaicin suggesting that this is a nonadrenergic-noncholinergic vasodilation and is a neurogenic response. The vasodilation was abolished by anti-serum against calcitonin-gene related peptide (CGRP) in a concentration-dependent manner. These data suggest that the non-adrenergic-noncholinergic vasodilation is mediated by endogenous CGRP released from the primary sensory nerve terminals upon electrical stimulation. In addition to the vasodilator action, CGRP also inhibited nerve stimulation-induced and norepinephrine-induced vasoconstriction at extremely low concentrations. The inhibitory action of CGRP appeared to be mediated by postsynaptic mechanisms inasmuch as evoked norepinephrine release was not affected by CGRP when the vasoconstriction produced by norepinephrine or periarterial nerve stimulation was attenuated greatly by CGRP. These observations suggest that the vascular tone of the resistance vessels can be regulated by primary sensory nerve-derived CGRP.

Inhibition of periarterial nerve stimulation-induced vasodilation of the mesenteric arterial bed by CGRP (8-37) and CGRP receptor desensitization.

We provide the first functional evidence that calcitonin gene-related peptide (8-37) induces a direct vasoconstriction and reversibly antagonizes vasodilation of the mesenteric arterial bed induced by calcitonin gene-related peptide (CGRP) suggesting that CGRP (8-37) is a competitive antagonist of vascular CGRP receptors. Vasodilation induced by periarterial nerve stimulation was inhibited both by CGRP (8-37) and by desensitization of CGRP receptors. These results further support the evidence that the periarterial nerve stimulation-induced nonadrenergic noncholinergic vasodilation of the mesenteric vasculature is mediated by endogenous CGRP and its receptors.

Neuropeptides in hypertension: role of neuropeptide Y and calcitonin gene related peptide.

1. The effect of neuropeptide Y (NPY) on cardiovascular function at three levels of the noradrenergic axis where the peptide is known to co-exist with noradrenaline (NA) and or adrenaline (A) was studied in normotensive Sprague-Dawley (SD), Wistar-Kyoto (WKY) or spontaneously hypertensive rats (SHR). 2. In the perfused mesenteric arterial bed, NPY and the structurally similar peptide intestinal polypeptide (PYY) decreased the periarterial nerve stimulation induced release of NA and potentiated the increase in perfusion pressure to nerve stimulation or exogenously applied agonists (e.g. angiotensin, vasopressin, phenylephrine). In contrast to NPY and PYY, C-terminal NPY fragments inhibited NA release and produced a parallel decrease in perfusion pressure thus supporting the concept of Y1 (post) and Y2 (pre) NPY receptors. 3. In the mesenteric artery of SHR the prejunctional inhibitory effect of NPY was attenuated while the postjunctional response was enhanced. 4. Following intrathecal (Int) injection of NPY, there was a decrease in blood pressure, total peripheral resistance (predominantly by a decrease in mesenteric vascular resistance) and renal nerve activity. The depressor effect of Int NPY was attenuated in the SHR. 5. Unilateral injections of NPY into the posterior hypothalamic nucleus increased blood pressure, hindquarter and renal vascular resistance and renal nerve activity. The pressor effect was enhanced in the SHR. 6. Periarterial nerve stimulation of the perfused mesenteric artery produced a frequency dependent vasodilation in beds pretreated with guanethidine and precontracted with methoxamine.(ABSTRACT TRUNCATED AT 250 WORDS)

Cardiovascular effects and modulation of noradrenergic neurotransmission following central and peripheral administration of neuropeptide Y.

Experiments have been conducted to evaluate the effect of neuropeptide Y (NPY) administered at three distinct levels of the nervous system: 1) the posterior hypothalamic nucleus, 2) the spinal cord, and 3) the vascular noradrenergic neuroeffector junction. It was observed that NPY produced varying cardiovascular effects at these three distinct sites of the nervous system. Microinjections into the posterior hypothalamic nucleus resulted in an increase in blood pressure, which was reduced by prior microinjection of a muscarinic or H1-histamine antagonist but not an H2-histamine antagonist. In addition to the involvement of histaminergic and cholinergic pathways, the pressor effect of NPY appears to result from an increase in sympathetic outflow. NPY was also seen to decrease the potassium-induced release of norepinephrine (NE) from slices obtained from the posterior hypothalamic nucleus. In contrast to what was observed in the hypothalamus, the intrathecal injection of NPY at a level of T4 or T10 in anesthetized or T10 in unanesthetized rats resulted in a depressor effect as well as a decrease in heart rate. Both an alpha 2- and beta-adrenoceptor antagonist reduced the NPY effect. The depressor effect of intrathecal NPY was attenuated in rats pretreated with reserpine as well as in Spontaneously Hypertensive rats (SHR). These data suggest that the effects of NPY are closely associated with sympathetic preganglionic neurons in the spinal cord. At the vascular noradrenergic neuroeffector junction, NPY decreased the nerve stimulation-induced release of NE while potentiating the contractile response. Moreover, NPY potentiated the increase in perfusion pressure of the perfused mesenteric arterial bed in response to angiotensin, vasopressin, or phenylephrine.(ABSTRACT TRUNCATED AT 250 WORDS)

Prejunctional and postjunctional effects of neuropeptide Y at the noradrenergic neuroeffector junction of the perfused mesenteric arterial bed of the rat.

The effect of neuropeptide Y (NPY) on periarterial nerve stimulation-induced release of norepinephrine (NE) and increase in perfusion pressure in the perfused mesenteric arterial bed of the rat was examined. Perfusate effluents were continuously collected and assayed for endogenous NE by high-pressure liquid chromatography (HPLC) coupled to electrochemical detection. Perfusion pressure was continuously monitored by means of a pressure transducer. Periarterial nerve stimulation (8 or 16 Hz, 60 V, 2-ms duration for 30 s) resulted in a readily detectable increase in NE release and perfusion pressure that was attenuated by the prior administration of tetrodotoxin (TTX) (10(-5) M) or guanethidine (5 X 10(-5) M). NPY exerted both prejunctional and postjunctional effects on noradrenergic neurotransmission in this preparation. The peptide produced a concentration-dependent reduction in the release of NE over a concentration range of 10(-10) - 10(-7) M. A similar inhibition effect occurred at 8, 10, and 16 Hz. In contrast, low concentrations (10(-10) and 10(-9) M) decreased the effect of nerve stimulation on perfusion pressure, whereas higher concentrations (10(-7) M) produced a marked potentiation. The alpha 2-adrenoceptor antagonist, yohimbine, did not alter the inhibitory effect of NPY on evoked NE release or the effect on perfusion pressure. Prazosin similarly did not alter the inhibitory effect of NPY on NE release but prevented the increase in perfusion pressure. We conclude that NPY modulates noradrenergic neurotransmission in the mesenteric arterial bed by decreasing the evoked release of NE and producing a concentration-dependent biphasic response on vascular smooth muscle.(ABSTRACT TRUNCATED AT 250 WORDS)

Preferential stimulation of ventral tegmental area dopaminergic neurons by nicotine.

The effect of intravenous (i.v.) nicotine on the single unit activity of midbrain dopamine (DA) neurons was studied in rats under either local or general anesthesia. Nicotine (50-500 micrograms/kg) produced a dose-related increase in the firing rate of nigral pars compacta DA cells (A9), up to 25% above baseline, irrespective of the preparation. The same range of doses was more than three times as effective on ventral tegmental area DA cells (A10) in rats paralyzed and given a local anesthetic. By contrast, the majority of these cells were temporarily depressed in deeply anesthetized animals. All of the above effects were reversed and prevented by i.v. mecamylamine suggesting the involvement of nicotine cholinergic receptors. Moreover, after nicotine-induced stimulation, low doses of i.v. apomorphine inhibited the firing rate similar to controls indicating that dopamine receptors are not directly involved in the nicotinic action. The results suggest that acute nicotine shares with other drugs of abuse the characteristic of being more effective in stimulating A10 than A9 neurons.

Alterations in the field stimulation-induced release of endogenous norepinephrine from the coccygeal artery of spontaneously hypertensive and Wistar-Kyoto rats.

The field stimulation-induced release of endogenous NE from the isolated caudal artery from 5-6, 8-10 and 28-30 week old SHR resulted in a greater release of transmitter compared to age-matched WKY. The alpha 2-selective adrenoceptor antagonist, yohimbine produced a significant enhancement in the release of NE from both SHR and WKY of 5-6 and 10-12 and 28 week old WKY but release was attenuated in 28 week old SHR. It is concluded that the enhanced release of NE contributes to the development of hypertension in the SHR.

Alterations in the release of norepinephrine at the vascular neuroeffector junction in hypertension.

The field stimulation induced release of 3H-norepinephrine (NE) from the isolated portal vein and endogenous NE from the isolated caudal artery and perfused mesenteric arterial bed of spontaneously hypertensive rats (SHR) and age-matched normotensive rats (Wistar-Kyoto or Sprague-Dawley) was studied. There was a significantly greater release of NE from all three preparations obtained from 10- to 12-week-old SHR compared to normotensive animals. In addition, there was a greater release of NE from the caudal artery of 5- to 6-week-old SHR compared to controls. No differences were seen in the evoked release of NE from portal vein or caudal artery obtained from renal or DOCA salt hypertensives compared to vessels obtained from sham controls. Neuropeptide Y (NPY) produced a concentration-dependent decrease in the field stimulation induced release of NE from the perfused mesenteric artery. Low concentrations of NPY decreased while higher concentrations potentiated the increase in perfusion pressure. The NPY induced inhibition of evoked NE release was not altered by alpha 1- or alpha 2-adrenoceptor antagonists while the alpha 1-adrenoceptor antagonist, prazosin, prevented the postjunctional response. These results are consistent with there being an alteration of NE release at the vascular neuroeffector junction in SHR which may contribute to the development or maintenance of hypertension. NPY exerts a modulatory role in noradrenergic transmission at the vascular neuroeffector junction.

Comparison of norepinephrine release in hypertensive rats: II. Caudal artery and portal vein.

It was observed that there was a significantly greater field-stimulation induced release of norepinephrine from the portal vein and caudal artery obtained from 28 week old SHR compared to WKY. The greater field-stimulation induced release of norepinephrine observed in the blood vessels of the SHR compared to WKY was not seen in DOCA-salt and one kidney-one clip hypertensive animals with similar elevations of systolic blood pressure. It was also observed that there was an attenuation of the effect of the prejunctional alpha 2-adrenoceptor antagonist, yohimbine to enhance the field-stimulation induced release of norepinephrine from blood vessels of the 28 week old SHR. There was no attenuation of the yohimbine effect in the other two models of hypertension. It is concluded that there is an alteration in the release of norepinephrine from blood vessels of SHR, resulting in a greater evoked release of the transmitter. A decrease in the functional activity of prejunctional alpha 2-adrenoceptor may contribute to the enhanced release of norepinephrine from the blood vessels of the SHR.

The effect of opioid drugs on the release of dopamine and 5-hydroxytryptamine from rat striatum following activation of nicotinic-cholinergic receptors.

The effect of (Met5)enkephalin, (D-Ala2,D-Met5)enkephalin, (Leu5)enkephalin, (D-Ala2,D-Met5)enkephalin and morphine on the release of [3H]dopamine, endogenous dopamine and [3H]5-hydroxytryptamine produced by the nicotinic-cholinergic agonist, dimethylphenyl piperazinium iodide (DMPP), was examined in rat striatal slices. The DMPP-induced release of [3H]dopamine and endogenous dopamine was reduced by the presence of (Met5)enkephalin, (D-Ala2,D-Met5)enkephalin (1-10 microM) or morphine (10 microM) but not by (Leu5)enkephalin or (D-Ala2,D-Leu5)enkephalin. The DMPP-induced release of [3H]5-hydroxytryptamine was reduced by (Leu5)enkephalin, (D-Ala2,D-Leu5)enkephalin, (Met5)enkephalin, (D-Ala2,D-Leu5)enkephalin (1-10 microM), and morphine (10 microM). All three opioids failed to alter the release of [3H]dopamine induced by field stimulation or potassium depolarization (30 microM). The inhibitory effects of opioid peptides and morphine demonstrated in the present study appear to be due to an initial interaction with nicotinic-cholinergic receptors in the striatum.

Relative potency of dopamine agonists on autoreceptor function in various brain regions of the rat.

The effect of six dopamine agonists including apomorphine, epinine, dopamine, piribedil, lergotrile and bromocriptine on the incorporation of [3H]tyrosine into dopamine was studied in slices and synaptosomes prepared from various brain areas containing dopamine terminals including striatum, nucleus accumbens, olfactory tubercle and medial basal hypothalamus. It was observed that all of these drugs were active in causing a decrease in dopamine synthesis in these various brain areas. The catecholamine agonists apomorphine, epinine and dopamine were more potent in inhibiting dopamine synthesis in the mesolimbic structures than in the striatum. On the other hand, apomorphine and epinine were less potent while dopamine was more potent in the medial basal hypothalamus. The ergoline drugs were weak agonists in all structures studied. It is concluded that autoreceptor regulation of dopamine synthesis is more active in the mesolimbic compared with the nigrostriatal dopamine pathway while autoreceptors may be absent in the median eminence.