neuropeptide-y and alpha-beta-methyleneadenosine-5--triphosphate

ATP, norepinephrine and NPY are co-released by sympathetic nerves innervating arteries. ATP elicits vasoconstriction via activation of smooth muscle P2X receptors. The functional interaction between neuropeptide Y (NPY) and P2X receptors in arteries is not known. In this study we investigate the effect of NPY on P2X1-dependent vasoconstriction in mouse mesenteric arteries. Suramin or P2X1 antagonist NF449 abolished α,β-meATP evoked vasoconstrictions. NPY lacked any direct vasoconstrictor effect but facilitated the vasoconstrictive response to α,β-meATP. Mesenteric arteries expressed Y

Topics: Adenosine Triphosphate; Adrenergic alpha-1 Receptor Antagonists; Animals; Benzenesulfonates; Calcium Channel Blockers; Calcium Channels, L-Type; Male; Mesenteric Arteries; Mice, Inbred C57BL; Neuropeptide Y; Nifedipine; Prazosin; Receptors, Neuropeptide Y; Receptors, Purinergic P2X1; Suramin; Sympathetic Nervous System; Vasoconstriction

The periarterial electrical nerve stimulation (30 s trains of pulses at a frequency of 1, 4 or 10 Hz) induced a double peaked vasoconstriction consisting of an initial transient constriction (first peak) followed by a prolonged response (second peak) in the isolated, perfused canine splenic artery. At low frequencies (1 and 4 Hz), a neuropeptide Y (NPY) Y(1) receptor antagonist BIBP 3226 (0.1-1 microM) produced a dose-dependent inhibitory effect on the second peak, but did not modify the first peak. At a high frequency (10 Hz), 1 microM BIBP 3226 induced a slight, but significant inhibition on both the first and second peaked responses. At a low frequency (1 Hz), the first peak was not influenced by blockade of alpha(1)-adrenoceptors or NPY Y(1) receptors with prazosin (0.1 microM) or BIBP 3226 (1 microM), respectively, but abolished by P2X receptor desensitization with alpha,beta-methylene ATP (alphabeta-m ATP, 1 microM). At a high frequency (10 Hz), the first peak was mostly inhibited by alphabeta-m ATP and partially by prazosin and BIBP 3226. On the other hand, the second peak at a low frequency was largely decreased by BIBP 3226 and partially by prazosin and alphabeta-m ATP, whereas at a high frequency, it was largely attenuated by prazosin and partially by alphabeta-m ATP and BIBP 3226. The results suggest that at a low frequency, the firstly transient constriction of double peaked responses is mainly induced via an activation of P2X-receptors, whereas at a high frequency, it is mostly mediated by the P2X-receptors, and partially by alpha(1)-receptors and NPY Y(1)-receptors. The secondary prolonged vasoconstriction at frequencies used is predominantly mediated via both alpha(1)-receptor and NPY Y(1) receptor activations, and in part by P2X-receptors. Furthermore, an activation of NPY Y(1) receptors may play an important role in evoking the prolonged vasoconstrictor response to longer pulse trains of stimulation at a low frequency, whereas an alpha(1)-adrenoceptor activation exerts a main vasomotor effect for the prolonged response at a high frequency.

Topics: Adenosine Triphosphate; Adrenergic alpha-Antagonists; Animals; Anti-Anxiety Agents; Arginine; Dogs; Drug Interactions; Electric Stimulation; Female; In Vitro Techniques; Male; Nerve Tissue; Neuropeptide Y; Norepinephrine; Prazosin; Receptors, Neuropeptide Y; Splenic Artery; Vasoconstriction

This study has characterized constrictions of small cutaneous arteries in the guinea pig ear in response to electrical stimulation of the cervical sympathetic nerve (SNS) in vivo. Video microscopy and on-line image analysis were used to examine diameter changes of ear arteries (80-140 micrometers resting diameter) in anesthetized guinea pigs. Trains of 50-300 impulses, but not single pulses or short trains, produced frequency-dependent (2-20 Hz) constrictions. The purinoceptor antagonist suramin (30 microM) greatly reduced constrictions produced by exogenous ATP but did not affect constrictions produced by SNS at 10 Hz or exogenous norepinephrine. The alpha(2)-adrenoceptor antagonist yohimbine (1 microM) enhanced the peak amplitude of sympathetic constrictions at lower stimulation frequencies (1-5 Hz). The amplitude of constrictions to SNS at 10 Hz was reduced, and the latency of constrictions was increased by the alpha(1)-adrenoceptor antagonist prazosin (1 microM). Constrictions to SNS at 10 Hz remaining after prazosin treatment were reduced in amplitude by dihydroergotamine (2 microM) and were attenuated further by the neuropeptide Y Y(1)-receptor antagonist 1229U91 (0.3 microM). Thus norepinephrine and neuropeptide Y act as cotransmitters to mediate sympathetic constriction of small ear arteries at higher stimulation frequencies (10 Hz), but ATP does not seem to contribute directly to these constrictions.

Topics: Adenosine Triphosphate; Animals; Arteries; Constriction, Pathologic; Electric Stimulation; Female; Guinea Pigs; Male; Neurons; Neuropeptide Y; Norepinephrine; Peptides, Cyclic; Skin; Suramin; Sympathetic Nervous System; Vasoconstriction

Elevated levels of nerve growth factor (NGF) protein and NGF mRNA have been reported in the vessels of spontaneously hypertensive rats (SHR: hypertensive, hyperactive) compared to Wistar-Kyoto (WKY) rats. Elevated NGF may be involved in the development of hypertension in SHRs. We examined vascular NGF mRNA and protein content and the regulation of NGF secretion by vascular smooth muscle cells (VSMCs) from two inbred strains (WKHT: hypertensive; WKHA: hyperactive) derived from SHRs and WKYs. Our goal was to determine if receptor-mediated defects in NGF regulation play a role in increased secretion of VSMC NGF from hypertensive animals. Tissue NGF mRNA content was determined by competitive, quantitative RT-PCR. Tissue NGF and NGF content in cultured VSMC-conditioned medium was quantified using a two-site ELISA. Tail artery NGF mRNA was elevated in WKHTs compared to WKHAs. Tissue NGF protein was elevated in WKHT aorta, mesenteric, and tail artery compared to WKHAs. Pharmacologically induced increases in NGF output were blocked with inhibition of transcription or protein synthesis. Basal NGF secretion by WKHT VSMCs was significantly higher than WKHAs. The observed increases in VSMC NGF output in SHRs over WKYs in response to beta-adrenergic agents are not preserved in the WKHT:WKHA comparison. Protein kinase C-dependent increases in SHR VSMC NGF appear in both WKHTs and WKHAs. In contrast, elevated NGF levels due to disturbances in alpha-adrenergic, peptidergic, and purinergic control of NGF output are features common to both genetic models of hypertension (SHR and WKHT). These results suggest that the defect in smooth muscle NGF metabolism observed in SHRs cosegregates with a hypertensive rather than a hyperactive phenotype. Moreover, altered receptor-mediated regulation (alpha-adrenergic, peptidergic, and purinergic) of VSMC NGF production may contribute to elevated vascular tissue NGF, suggesting a mechanism leading to the high levels of NGF associated with hypertension in SHRs and WKHTs.

Topics: Adenosine Triphosphate; Adrenergic Agents; Animals; Aorta, Thoracic; Cell Count; Cells, Cultured; Cholinergic Agents; Cyclic AMP-Dependent Protein Kinases; Cycloheximide; Dactinomycin; Female; Male; Mesenteric Arteries; Muscle, Smooth, Vascular; Nerve Growth Factors; Neuropeptide Y; Protein Kinase C; Rats; Rats, Inbred SHR; Rats, Inbred WKY; RNA, Messenger; Tail; Transcription, Genetic; Vasoconstrictor Agents

Neuropeptide Y (NPY), noradrenaline (NA) and ATP are cotransmitters of the sympathetic nervous system and exert vasocontractile effects. The aim of this study was to determine the role of these sympathetic co-transmitters in human hypertension. Subcutaneous vessels from 12 patients with essential hypertension and 12 matched controls were studied in vitro. Vascular contractile responses to NPY, NA, alpha,beta-methylene ATP (alpha,beta-mATP) and potassium were studied in isolated arteries and veins (diameter 0.1-1.1 mm) with intact endothelium. The dilatory effect of acetylcholine was used to test the endothelial function. There was no difference in potency (pD2) or contractile response to NPY, NA or alpha,beta-mATP between hypertensive and control arteries. In veins, however, the contractile response to NPY was significantly reduced in hypertensives and the responses to NA were unchanged. Furthermore, the sensitivity (pD2) to alpha,beta-mATP was significantly reduced in veins from hypertensives. There was no difference in the dilatory response to acetylcholine between the hypertensives and the controls, neither in the arteries nor in the veins, indicating that the observed changes in vascular reactivity to NPY, NA and alpha,beta-mATP were not endothelium-dependent. In conclusion, the postjunctional contractile effect of NPY and sensitivity (pD2) to alpha,beta-mATP, co-transmitters of the peripheral sympathetic nervous system, are attenuated in veins in essential hypertension.

Topics: Acetylcholine; Adenosine Triphosphate; Aged; Aged, 80 and over; Female; Humans; Hypertension; In Vitro Techniques; Male; Middle Aged; Neuropeptide Y; Neurotransmitter Agents; Norepinephrine; Sympathetic Nervous System; Vasoconstriction; Vasoconstrictor Agents; Veins

The effects of the neuropeptide Y Y1 receptor antagonist SR 120107A (1-[2-[2-(2-naphtylsulfamoyl)-3-phenylpropionamido]-3-[4-[N- [4- (dimethylaminomethyl)-cis-cyclohexylmethyl]amidino]phenyl]propiony l] pyrrolidine, (S,R) stereoisomer) on sympathetic non-adrenergic vasoconstriction in a variety, of vascular beds were studied in reserpinized anesthetized pigs in vivo. The rapid vasoconstrictor response evoked by single impulse stimulation, in hind limb and nasal mucosa, was not affected by SR 120107A (1.5 mg kg-1 i.v.). In contrast, SR 120107A potently inhibited the long-lasting phase of vasoconstriction evoked by high frequency (60 impulses at 20 Hz) sympathetic nerve stimulation, in the main and deep femoral, the saphenous and the internal maxillary arteries, leaving merely the initial rapid peak of vasoconstriction in these vessels. Furthermore, the vasoconstrictor response was nearly abolished in the kidney and was attenuated in the spleen and main femoral artery, despite maintained neuropeptide Y overflow. The vasoconstrictor response evoked in the kidney by peptide YY, a neuropeptide Y Y1 and Y2 receptor agonist, was also nearly abolished in the presence of SR 120107A. This inhibitory effect on the response to exogenous agonist correlated well with the long-lasting inhibition of the response to nerve stimulation in the same tissue. The peptide YY-evoked vasoconstriction in the spleen was not altered by SR 120107A, in accordance with the view that the neuropeptide Y receptor population in this organ consists mainly of neuropeptide Y Y2 receptors. SR 120107A did not influence the vasoconstrictor effects of alpha, beta-methylene ATP (mATP) or phenylephrine in any of the tissues studied. We conclude that SR 120107A is a potent neuropeptide Y Y1 receptor antagonist with long duration of action in vivo. Endogenous neuropeptide Y acting on the neuropeptide Y Y1 receptor is likely to account for the long-lasting component of the reserpine-resistant sympathetic vasoconstriction upon high frequency stimulation in hind limb and nasal mucosa. Furthermore, the peak vasoconstriction in kidney, and to some extent in spleen, is also neuropeptide Y Y1 receptor mediated.

Topics: Adenosine Triphosphate; Animals; Electric Stimulation; Female; Femoral Artery; Hindlimb; Male; Maxillary Artery; Naphthalenes; Nasal Mucosa; Neuropeptide Y; Phenylephrine; Pyrrolidines; Receptors, Neuropeptide Y; Renal Circulation; Spleen; Swine; Sympathetic Nervous System; Vasoconstriction; Vasoconstrictor Agents

Vascular smooth muscle contractile responses to neuropeptide Y, alpha,beta-methyleneATP and noradrenaline were studied in circular segments of isolated vessels with intact endothelium in vitro from 12 patients with diabetes mellitus type 2 (NIDDM) and 12 control subjects. The dilatory effect of acetylcholine was used to test the function of the endothelium. Subcutaneous arteries and veins (diameter 0.1-1.1 mm) were obtained during surgery. There was no difference in contractile responses to noradrenaline or alpha,beta-methyleneATP between diabetic and control vessels. The contractile response to neuropeptide Y, however, was markedly reduced in the diabetic group. The maximal contractile effect (46.0 +/- 14.0%, p < 0.05) but not the sensitivity to neuropeptide Y was significantly less in diabetic veins compared to control (107.5 +/- 19.6%). Thus, the attenuation of neuropeptide Y responses was present in humans as previously observed in alloxan-induced diabetes mellitus in rabbits. There was no difference in the dilator effect of acetylcholine between the diabetic and the control group in any of the vessel types, indicating that the difference in vascular reactivity to neuropeptide Y was not endothelium-dependent. In conclusion, the present study has shown that the postjunctional effects of neuropeptide Y, a co-transmitter of the peripheral sympathetic nervous system, is selectively attenuated in diabetes mellitus.

Topics: Acetylcholine; Adenosine Triphosphate; Aged; Aged, 80 and over; Diabetes Mellitus, Type 2; Female; GTP-Binding Proteins; Humans; In Vitro Techniques; Male; Middle Aged; Muscle Contraction; Muscle, Smooth, Vascular; Neuropeptide Y; Norepinephrine; Potassium; Vasoconstriction; Vasoconstrictor Agents

Neuropeptide Y(NPY) inhibits Ca2+-activated K+ channels reversibly in vascular smooth muscle cells from the rat tail artery. NPY (200 microM) had no effect in the absence of intracellular adenosine 5'-triphosphate (ATP) and when the metabolic poison cyanide-M-chlorophenyl hydrozone (10 microM) was included in the intracellular pipette solution. NPY was also not effective when ATP was substituted by the non-hydrolysable ATP analogue adenosine 5'-[beta gamma-methylene]-triphosphate (AMP-PCP). NPY inhibited Ca2+-activated K+ channel activity when ATP was replaced by adenosine 5'-O-(3-thiotriphosphate) (ATP [gamma-S]) and the inhibition was not readily reversed upon washing. Protein kinase inhibitor (1 microM), a specific inhibitor of adenosine 3', 5'-cyclic monophosphate-dependent protein kinase, had no significant effect on the inhibitory action of NPY. The effect of NPY on single-channel activity was inhibited by the tyrosine kinase inhibitor genistein (10 microM) but not by daidzein, an inactive analogue of genistein. These observations suggest that the inhibition by NPY of Ca2+-activated K+ channels is mediated by ATP-dependent phosphorylation. The inhibitory effect of NPY was antagonized by the tyrosine kinase inhibitor genistein.

Topics: Adenosine Triphosphate; Animals; Arteries; Calcium; Enzyme Inhibitors; Genistein; In Vitro Techniques; Isoflavones; Male; Membrane Potentials; Muscle, Smooth, Vascular; Neuropeptide Y; Patch-Clamp Techniques; Potassium Channels; Protein-Tyrosine Kinases; Rats; Rats, Wistar

1. Immunohistochemical and isolated organ bath techniques were used to detect the presence of neuropeptide Y (NPY) in the rat urinary bladder and to determine its effect on tone, spontaneous activity and contractile responses of the detrusor muscle to electrical field stimulation, acetylcholine and alpha,beta-methylene ATP (alpha,beta-MeATP). 2. A very rich presence of NPY-immunoreactive nerve fibres was found mainly within the bundles of detrusor muscle cells. Chronic treatment with 6-hydroxydopamine did not affect the density of NPY-positive nerve fibres. 3. NPY (> 1 nM) enhanced the force and frequency of spontaneous contractions and generated a rise in the resting tone of the detrusor. These effects of NPY on the tone and the spontaneous activity remained unaffected by atropine (3 microM), indomethacin (10 microM) and aspirin (100 microM) but were abolished by Ca(2+)-withdrawal from the bathing medium. 4. The enhancing effects of NPY on the spontaneous contractions and the resting tone were not prevented by the induction of purinoceptor desensitization. 5. NPY (1-250 nM) potentiated electrical field stimulation (EFS, 1-64 Hz, 0.1 ms pulses duration, 10s train duration)-evoked, tetrodotoxin (0.5 microM)-sensitive contractions. The atropine (3 microM)-resistant component of EFS-evoked contractions was also potentiated by NPY. By contrast, the nifedipine (1 microM)-resistant but atropine-sensitive component of EFS-evoked contraction was inhibited by NPY. 6. NPY (250 nM) did not affect acetylcholine-evoked contractions, but potentiated alpha,beta-MeATP-evoked contractions. 7. It is concluded that NPY-innervation of rat urinary bladder is largely confined to the detrusor muscle and is abundant and mainly non-adrenergic. It is further concluded that the enhancing effect of NPY on detrusor spontaneous activity and tone is caused by Ca2+ influx through nifedipine-sensitive Ca2+ channels and is not mediated through acetylcholine or cyclo-oxygenase-sensitive eicosanoids or ATP.8. The results are consistent with the hypothesis that intrinsic NPY in the rat detrusor innervation contributes to the motor transmission in two ways: by promoting non-cholinergic motor transmission and by inhibiting prejunctionally the cholinergic transmission.

Topics: Acetylcholine; Adenosine Triphosphate; Animals; Autonomic Nervous System; Calcium; Electric Stimulation; Immunohistochemistry; In Vitro Techniques; Male; Muscle Contraction; Muscle Tonus; Muscle, Smooth; Neuropeptide Y; Nifedipine; Oxidopamine; Rats; Rats, Wistar; Synaptic Transmission; Urinary Bladder

Electrical transmural stimulation of isolated iris dilator muscle of the rabbit produced a transient contraction that consisted of adrenergic and nonadrenergic components. In contrast to the adrenergic component, the nonadrenergic component was resistant to prazosin and other adrenoceptor antagonists. However, both components were completely blocked by guanethidine or tetrodotoxin. Among some tested compounds including neuropeptide Y, both ATP and 2-methylthio ATP produced a transient contraction in the dilator muscle and the sustained treatment with each markedly attenuated the nonadrenergic responses to electrical stimulation and to ATP. Suramin had no effect on and alpha,beta-methylene ATP potentiated the responses to electrical stimulation and to ATP. These results suggest that the nonadrenergic contraction induced by electrical transmural stimulation is a sympathetic purinergic response that may be mediated through unique purinoceptors.

Topics: Adenosine Triphosphate; Adrenergic alpha-Antagonists; Animals; Electric Stimulation; Female; Guanethidine; In Vitro Techniques; Iris; Male; Muscle Contraction; Neuropeptide Y; Prazosin; Rabbits; Receptors, Purinergic; Suramin; Tetrodotoxin; Thionucleotides

1. Strips of human saphenous veins were superfused with Krebs-Henseleit solution at either 25 degrees C or 37 degrees C. Constrictor responses to electrical stimulation (10 Hz, 40 s) but not to exogenous noradrenaline (0.1, 1 microM) were abolished by guanethidine (10 microM) and tetrodotoxin (1 microM). Hence, responses to electrical stimulation are due to action potential-induced release of sympathetic neurotransmitters. 2. Constrictor responses to electrical stimulation and noradrenaline were reduced by the alpha 1-adrenoceptor antagonist, prazosin (0.3 microM) as well as by the alpha 2-adrenoceptor antagonist, rauwolscine (1 microM). The combination of prazosin and rauwolscine abolished constrictor responses to noradrenaline at 25 degrees C and 37 degrees C. However, constrictor responses to electrical stimulation were partly resistant to alpha-adrenoceptor blockade by prazosin and rauwolscine (at 25 degrees C about 30%). Residual constrictor responses to electrical stimulation were also observed in the presence of the combination of prazosin (3 microM) and rauwolscine (10 microM) as well as in the presence of phenoxybenzamine (10 microM). 3. Veins, incubated with [3H]-noradrenaline, released tritium upon electrical stimulation (10 Hz, 40 s). Moreover, electrical stimulation also induced an overflow of ATP amounting to 4.8 +/- 1.5 pmol g-1 at 25 degrees C and 2.0 +/- 0.5 pmol g-1 at 37 degrees C. Both tritium and ATP overflow were abolished by tetrodotoxin (0.5 microM). The combination of prazosin (0.3 microM) and rauwolscine (1 microM) increased tritium overflow at either 25 degrees C or 37 degrees C by about 120%, but reduced ATP overflow by about 70%. Hence, a significant percentage of the electrically evoked ATP overflow seems to be released from non-neuronal cells upon activation of alpha-adrenoceptors by endogenous noradrenaline. The remaining ATP overflow, which was resistant to alpha-adrenoceptor blockade, may reflect neuronally released ATP.4. ATP (300 MicroM) and alpha,Beta-methylene-ATP (1, 10 MicroM), both induced constrictor responses. The P2-purinoceptor antagonist, suramin (300 MicroM) markedly inhibited constrictor responses to ATP and alpha, beta-methylene-ATP, but not those to electrical stimulation and to noradrenaline. Moreover, suramin(300 MicroM) failed to diminish the alpha-adrenoceptor blockade-resistant constrictor response to 10 Hz.5. In conclusion, constrictor responses to sympathetic nerve stimulation in human saphenous vei

Topics: Action Potentials; Adenosine Triphosphate; Adrenergic alpha-Antagonists; Adult; Aged; Aged, 80 and over; Electric Stimulation; Female; Guanethidine; Humans; In Vitro Techniques; Male; Middle Aged; Neuropeptide Y; Norepinephrine; Receptors, Purinergic; Saphenous Vein; Suramin; Synaptic Transmission; Tetrodotoxin; Vasoconstriction

The acute influence of exogenous transmitters and sympathetic nerve stimulation on the composition of blood cells in the splenic vein in relation to the splenic vascular effects was investigated in anaesthetized pigs. Intra arterial bolus injections of 720 pmol neuropeptide Y (NPY), 4.9 nmol noradrenaline (NA) and 20 nmol alpha,beta-methylene adenosine triphosphate (mATP) in the spleen were given and these doses caused arterial vasoconstriction in the same range, and increase in splenic venous haematocrit. NPY administration evoked a decrease in splenic venous blood flow and an unchanged leukocyte outflow from the spleen. mATP and NA, on the other hand, evoked increases in splenic venous blood flow and leukocyte outflow. Sympathetic nerve stimulation caused increases in haematocrit and leukocyte outflow in control pigs as well as in pigs with reserpine-induced depletion of tissue NA, although these effects, as well as the vascular effects, were significantly reduced after reserpine treatment. For comparison, the vasodilator calcitonin gene-related peptide increased leukocyte outflow without change in haematocrit. It is concluded that haematocrit and leukocyte concentration in the splenic venous blood are acutely modulated in different ways by vascular changes evoked by different sympathetic mediators. Furthermore, the capacitance function seems to be regulated by adrenergic and possibly purinergic transmission, whereas the non-adrenergic mediator NPY seems to be involved mainly in splenic arterial vasoconstriction.

Topics: Adenosine Triphosphate; Animals; Binding Sites; Blood Cells; Calcitonin Gene-Related Peptide; Electric Stimulation; Female; Hematocrit; Leukocyte Count; Leukocytes; Male; Neuropeptide Y; Norepinephrine; Regional Blood Flow; Spleen; Splenic Vein; Swine; Sympathetic Nervous System

The role of the endothelium in the potentiating action of neuropeptide Y (NPY) to contraction induced by KCl, alpha, beta-methylene ATP (mATP), and noradrenaline (NA) was tested on rat tail arteries. Endothelium-intact and denuded ring segments and freshly isolated single smooth muscle cells were used in the study. Contraction responses to KCl and mATP were potentiated by NPY (50 nM) in both intact and denuded arteries. Contraction to NA was potentiated by NPY at 500 nM but not at 50 nM. The potentiation effect of NPY was antagonized by nifedipine. Similarly, the shortening of single smooth muscle cells in response to KCl and mATP was potentiated by NPY (50 nM). The noradrenaline response was potentiated by NPY at 500 nM but not at 50 nM. Our results suggest that the potentiating effect of NPY is more specific to contraction mediated by nifedipine-sensitive calcium channels and is not dependent on the presence of an intact endothelium.

Topics: Adenosine Triphosphate; Animals; Arteries; Calcium Channels; Drug Synergism; Endothelium, Vascular; In Vitro Techniques; Muscle, Smooth, Vascular; Neuropeptide Y; Nifedipine; Norepinephrine; Potassium Chloride; Rats; Tail; Vasoconstriction

The question of the existence of postjunctional, contraction-mediating alpha 2-adrenoceptors, in addition to the known alpha 1-adrenoceptors, was studied in the mouse isolated vas deferens. Both the alpha 1-selective agonist phenylephrine and the alpha 2-selective agonist 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline (UK 14,304) caused contraction of the vas deferens. In the presence of the alpha 1-selective antagonist prazosin (added in order to prevent an alpha 1 component in the effect of high concentrations of UK 14,304), the alpha 2-selective antagonist yohimbine and idazoxan shifted the concentration-response curve of UK 14,304 to the right in a manner compatible with competitive antagonism and with dissociation constants KB indicating the involvement of alpha 2-adrenoceptors. The maximal contraction elicited by UK 14,304 (in the presence of prazosin) was much lower than the maximal contraction elicited by phenylephrine. The effect of UK 14,304 was not changed by the P2-purinoceptor agonist alpha,beta-methylene-ATP and was reduced by neuropeptide Y, but was markedly enhanced by relatively low concentrations of phenylephrine. When the sympathetic fibres of the vas deferens were stimulated by trains of ten widely spaced (0.5 Hz) electric pulses, the tissue responded with ten separate twitches in which purinergic and adrenergic components were isolated by prazosin and suramin, respectively. Prazosin reduced the first adrenergic twitch in these trains at concentrations close to its KB value at alpha 1-adrenoceptors, whereas yohimbine and idazoxan reduced the first adrenergic twitch at concentrations far lower than their KB values at alpha 1-adrenoceptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenosine Triphosphate; Adrenergic alpha-Antagonists; Animals; Brimonidine Tartrate; Dioxanes; Drug Interactions; Idazoxan; Male; Mice; Muscle Contraction; Neuropeptide Y; Phenylephrine; Prazosin; Quinoxalines; Receptors, Adrenergic, alpha; Suramin; Vas Deferens; Yohimbine

The effect of acrylamide intoxication on the innervation and local control of the rabbit central ear artery was investigated. There was no difference in the noradrenaline, neuropeptide Y and calcitonin gene-related peptide tissue content between control and experimental animals. There was, however, a slight reduction in catecholamine histofluorescence. Although the contractile efficiency of the rabbit central ear artery as measured by responses to potassium chloride was unchanged, nerve-mediated contractile responses were significantly attenuated in acrylamide-treated animals. Contractile responses induced by exogenous alpha,beta-methylene ATP were markedly increased after acrylamide treatment, in contrast to contractions induced by exogenous noradrenaline which were attenuated at maximal concentrations. Modulatory effects of nerve-mediated contractile responses by neuropeptide Y were unaffected by acrylamide intoxication. It therefore appears that acrylamide intoxication damages sympathetic cotransmission, perhaps with preferential action on the purinergic component. Endothelium-dependent relaxant responses to acetylcholine and substance P were attenuated in acrylamide-treated animals, whereas relaxant responses mediated by calcitonin gene-related peptide (endothelium independent) were unaffected. The question of whether the damage to the endothelial cell action is a primary effect, or a secondary consequence of sympathetic nerve damage, is discussed.

Topics: Acrylamide; Acrylamides; Adenosine Triphosphate; Animals; Arteries; Ear; Endothelium, Vascular; Male; Neuropeptide Y; Norepinephrine; Rabbits; Sympathetic Nervous System; Vasoconstriction; Vasodilation

Contraction of the guinea pig saphenous artery induced by stimulation of perivascular nerves consists of an adrenergic and a purinergic component. Neuropeptide Y (NPY) potentiated the neural responses significantly at low stimulation frequencies but not at high frequencies when the contraction was maximal. After blocking the adrenoceptors with phentolamine, significant potentiation of the purinergic component by NPY could be demonstrated. The potentiating effect of NPY was abolished in the presence of nifedipine. After desensitizing the purinoceptors with alpha,beta-methylene-ATP, there was no potentiation of the remaining adrenergic component by NPY. NPY also has no effect on contractions induced by exogenous norepinephrine but potentiates those induced by alpha,beta-methylene-ATP. These results suggest that NPY potentiates neurally induced contraction in the guinea pig saphenous artery by a specific action on the purinergic response.

Topics: Adenosine Triphosphate; Animals; Arteries; Electric Stimulation; Guinea Pigs; In Vitro Techniques; Male; Neuropeptide Y; Norepinephrine; Receptors, Adrenergic; Receptors, Purinergic

We have studied the neurogenic response of small mesenteric arteries from the rat to evaluate the involvement of possible co-transmitters under various modes of stimulation. Segments of small branches of the mesenteric artery were mounted in a myograph and the intramural nerves were activated with transmural electrical stimulation. A single stimulation of the nerves caused a contraction that was reduced by only 20% in the presence of adrenergic blocking agents (prazosin or phenoxybenzamine), whereas the steady-state response to continuous nerve stimulation of high frequency was reduced by 90-95%. In contrast, all responses to applied noradrenaline in doses up to at least 1 mM were eliminated by phenoxybenzamine treatment. The stable ATP analogue, alpha,beta-methylene ATP, reduced the response to a single nerve stimulation by 70%, but reduced the contraction caused by continuous high-frequency nerve stimulation by only 10%. None of these agents affected the response to applied neuropeptide Y (NPY). The response of relaxed vessels to nerve stimulation was totally blocked by the combination of an adrenoceptor-blocking agent and alpha,beta-methylene ATP, although even in this situation a further neurogenic response could be revealed in vessels precontracted with vasopressin. Responses to either single stimuli or brief burst stimulations were potentiated after high-frequency stimulation. Both the adrenergic and non-adrenergic components were enhanced to roughly the same extent. Also the potentiated response was eliminated by the combined application of prazosin and alpha,beta-methylene ATP. The non-adrenergic transmitter in the sympathetic nerves of small arteries thus appears to be the dominant transmitter during low-frequency nerve stimulation, causing rapid but phasic activation. Noradrenaline is the most important transmitter for higher frequencies, exerting slower but sustained contractions. The post-stimulatory potentiation affects both the adrenergic and the non-adrenergic part of the neurogenic response.

Topics: Adenosine Triphosphate; Adrenergic alpha-Antagonists; Animals; Dose-Response Relationship, Drug; Drug Interactions; Electric Stimulation; In Vitro Techniques; Male; Mesenteric Arteries; Muscle Contraction; Muscle, Smooth, Vascular; Neuromuscular Junction; Neuropeptide Y; Norepinephrine; Rats; Rats, Inbred Strains; Synaptic Transmission

1. We examined the neuromodulatory effects of neuropeptide Y (NPY) on purinergic and adrenergic co-transmission in the guinea-pig vas deferens. 2. In superfused vas deferens preparations, NPY (0.3 microM) inhibited the stimulus-evoked overflow of both ATP and [3H]-noradrenaline ([3H]-NA) at 2 Hz, but only the stimulus-evoked release of [3H]-NA at 20 Hz. 3. Postjunctionally, NPY greatly enhanced responses to alpha,beta-methylene ATP and to a lesser extent to exogenous NA. 4. Preparations stimulated in organ baths showed frequency-dependent contractions to field stimulation. NPY abolished responses to field stimulation at low frequency and a small number of pulses. At high frequency (20 Hz), NPY abolished responses elicited by 10 pulses, inhibited responses by 50% at 20 pulses and had little effect on preparations stimulated for 240 pulses. 5. Our study suggests that NPY neuromodulates co-transmission in the vas deferens by inhibiting the release of ATP and NA and that these effects predominate over the postjunctional enhancement by NPY. These results also show that the physiological effect of NPY will be determined both by the frequency at which the nerves are discharging and the duration of their firing.

Topics: Adenosine Triphosphate; Animals; Electric Stimulation; Guinea Pigs; In Vitro Techniques; Male; Muscle, Smooth; Neuromuscular Junction; Neuropeptide Y; Norepinephrine; Sympathetic Nervous System; Synaptic Transmission; Tachyphylaxis; Vas Deferens

The dose-response curve for the vasopressor effect of alpha, beta-methylene ATP in pithed rats was influenced by four suramin-related drugs (each at 100 mumol/kg). The curve was shifted to the right by a factor of 8 by 'compound 3' and by a factor of 2 by two other derivatives, but was not affected by the fourth analogue. Compound 3 had no effect on the vasopressor response to noradrenaline or neuropeptide Y. In conclusion, compound 3 is a purine P2X receptor antagonist which is approximately as potent as suramin, and which, like suramin, does not exhibit antagonistic properties at alpha-adrenoceptors and neuropeptide y receptors.

Topics: Adenosine Triphosphate; Animals; Blood Pressure; Dose-Response Relationship, Drug; Male; Neuropeptide Y; Norepinephrine; Purinergic Antagonists; Rats; Rats, Inbred Strains; Suramin; Vasoconstrictor Agents

The localisation of neuropeptide Y in the rabbit central ear artery and its pharmacological action on this preparation were investigated. Immunohistochemical studies demonstrated the presence of neuropeptide Y-like immunoreactivity in the perivascular nerves supplying the rabbit ear artery. Forty-eight hours after treatment with reserpine (5 mg/kg i.p. 48 h and 3 mg/kg i.p. 24 h prior to the experiment) catecholamine fluorescence in the rabbit central ear artery was abolished and the neuropeptide Y-like immunoreactivity was substantially reduced, suggesting that noradrenaline and neuropeptide Y were colocalised in sympathetic nerves. Contractile responses to exogenous noradrenaline (1 microM) and alpha,beta-methylene ATP (1 microM) were both significantly potentiated following incubation with neuropeptide Y (0.3 microM); the degree of potentiation being similar for both agonists. Electrical field stimulation of the rabbit central ear artery (16 and 64 Hz) produced frequency-dependent contractile responses which were abolished by tetrodotoxin (1 microM) and which were significantly potentiated in the presence of neuropeptide Y (0.3 microM). The responses to stimulation at 16 Hz were enhanced to a greater extent than the responses at 64 Hz. After blocking the noradrenergic component of the neurogenic response with prazosin (1 microM), the residual purinergic component, at both 16 and 64 Hz, was significantly enhanced in the presence of neuropeptide Y. However, following desensitisation of the P2-purinoceptor with alpha,beta-methylene ATP, neuropeptide Y had no significant effect on the residual noradrenergic component.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenosine Triphosphate; Animals; Arteries; Ear, External; Electric Stimulation; Histocytochemistry; Immunohistochemistry; In Vitro Techniques; Male; Muscle Contraction; Muscle, Smooth, Vascular; Neuropeptide Y; Norepinephrine; Prazosin; Rabbits; Receptors, Purinergic; Reserpine; Sympathetic Nervous System

1. The effects of sympathetic nerve stimulation (evoked by recordings of authentic irregular vasoconstrictor nerve fibre discharge with average frequencies of 0.59, 2.0 and 6.9 Hz) on the perfusion pressure and the overflow of noradrenaline (NA) and neuropeptide Y-like immunoreactivity (NPY-LI) were investigated in the blood-perfused gracilis muscle of the dog in situ. 2. Nerve stimulation in the untreated control group evoked a frequency-dependent increase in perfusion pressure and overflow of NA. A significant overflow of NPY-LI was found at the highest frequency only. 3. In a separate group of animals, the sympathetic supply was unilaterally interrupted by preganglionic decentralization before the administration of reserpine (1 mgkg-1 i.v.) 24 h before the experiment. Reserpine reduced the NA content of the intact and decentralized gracilis and gastrocnemius muscle by 98-99%. Reserpine also induced a marked (80%) reduction of the muscular content of NPY-LI. The depletion of NPY-LI was, in contrast to that of NA, prevented by the decentralization, suggesting that nerve impulse activity was of primary importance for the reserpine-induced depletion of NPY-LI. 4. A slowly developing and long-lasting perfusion pressure increase was evoked by nerve stimulation, at 2.0 and 6.9 Hz after reserpine treatment. These responses were larger in the decentralized, as compared to the intact gracilis muscle and correlated with the nerve stimulation evoked overflow of NPY-LI (r = 0.79, P less than 0.001). Stimulation at 0.59 Hz caused vasoconstriction in the decentralized but not in the intact gracilis. 5. Administration of alpha,beta,-methylene adenosine triphosphate did not evoke an increase in perfusion pressure in the gracilis muscle of reserpine-treated animals. 6. In conclusion, a large perfusion pressure increase to sympathetic nerve stimulation occurs in the reserpine-pretreated skeletal muscle vasculature of the dog in vivo, providing that preganglionic decentralization has been performed. It is suggested that the released NPY-LI may mediate this vasoconstrictor response.

Topics: Adenosine Triphosphate; Animals; Dogs; Electric Stimulation; Female; Male; Muscles; Neuropeptide Y; Norepinephrine; Radioimmunoassay; Reserpine; Sympathetic Nervous System; Vasoconstriction

Concomitant release of noradrenaline (NA) (using tritium labelling), adenosine 5'-triphosphate (ATP) (using the luciferin-luciferase firefly technique) and neuropeptide Y (using the enzyme-linked immunosorbant assay) during electrical stimulation of the guinea-pig vas deferens has been demonstrated. In addition it has been shown: (1) that release of NA and ATP is unaffected following selective desensitization of the P2 (ATP)-purinoceptor by alpha, beta-methylene ATP and (2) that total block of contractile responses of the vas deferens to nerve stimulation by a combination of prazosin and alpha, beta-methylene ATP does not reduce the release of NA and ATP. These results are consistent with the hypothesis that NA, ATP and neuropeptide Y are released as co-transmitters or modulators in sympathetic nerves supplying the vas deferens.

Topics: Adenosine Triphosphate; Adrenergic Fibers; Animals; Electric Stimulation; Guinea Pigs; In Vitro Techniques; Male; Muscle Contraction; Neuropeptide Y; Norepinephrine; Prazosin; Tetrodotoxin; Vas Deferens