Segments Of Ear Artery Research Articles

Identification of alpha-1L adrenoceptor in rabbit ear artery.

The alpha-1L adrenoceptor (AR) was identified in rabbit ear artery by both functional and ligand binding studies. In functional studies using arterial rings, the contractile response to NS-49 [(R)-(-)-3'-(2-amino-1-hydroxyethyl)-4'-fluorometh-anesulfonanilide hydrochloride] (alpha-1A and alpha-1L AR-selective agonist) was competitively antagonized with low affinities by prazosin, RS-17053 [N-[2-(2-cyclopropylmethoxyphenoxy) ethyl]-5-chloro-alpha,alpha-dimethyl-1H-indole-3-ethamine hydrochloride], and 5-methylurapidil but with high affinities by tamsulosin and KMD-3213 [(-)-1-(3-hydroxypropyl)-5-[(2R)-2-([2-[(2,2,2-trifluoroethoxy)phenoxy]ethyl]amino)propyl]-2,3-dihydro-1H-indole-7-carboxamide]. In contrast, the response to noradrenaline (nonselective alpha-1 AR agonist) was inhibited noncompetitively by these antagonists (except 5-methylurapidil) with Schild slopes different from unity. These results suggest that the response to NS-49 was mediated predominantly via alpha-1L ARs, whereas the response to noradrenaline was produced through two distinct alpha-1 AR subtypes (presumably alpha-1B and alpha-1L ARs). In binding studies with intact segments of rabbit ear artery, [3H]KMD-3213 bound with high affinity (pKD=9.7) to alpha-1 ARs, which were subdivided by prazosin, RS-17053, and 5-methylurapidil into two subtypes (alpha-1A and alpha-1L ARs). In contrast, [3H]prazosin binding sites in ear artery segments (pKD = 9.8) were identified as alpha-1A and alpha-1B ARs. In conventional binding studies using isolated rabbit ear artery microsomal membranes, [3H]KMD-3213 binding sites were identified as alpha-1A ARs with high affinities for prazosin, RS-17053, and 5-methylurapidil. Our study indicates that an alpha-1L AR having a unique pharmacological profile coexists with alpha-1A and alpha-1B ARs in rabbit ear artery and can be identified either functionally or by binding studies using intact tissues but not microsomal membrane preparations.

Regional differences in the effect of oestrogen on vascular tone in isolated rabbit arteries.

The aim of the study was to assess how oestrogen acutely affects the tone of isolated artery segments from different vascular beds in rabbit. Cumulative concentrations of 17beta-oestradiol were added to ring segments of thoracic aorta, pulmonary artery, ear artery and coronary arteries from adult male rabbits. Coronary arteries precontracted with potassium or the thromboxane agonist, U46619, relaxed to oestrogen (10(-7) to 10(-4) M), whereas oestrogen (10(-8) to 10(-4) M) only caused additional contraction in segments of thoracic aorta and pulmonary artery precontracted with phenylephrine. In the thoracic aorta both the phospholipase C inhibitor NCDC (10(-4) M) and the cyclooxygenase inhibitor indomethacin (10(-5) M) almost completely blocked the contractile effect of oestrogen. In segments of the ear artery, oestrogen caused relaxation only at higher concentrations of oestrogen (10(-5) to 10(-4) M). In conclusion, oestrogen may cause both relaxation and vasoconstriction in different vascular beds, and in the thoracic aorta the contractile effects of oestrogen may be mediated via inositol phosphate-dependent pathways and release of prostaglandins.

A study of NPY-mediated contractions of the porcine isolated ear artery.

Enhanced contractions of the porcine isolated ear artery by the alpha2-adrenoceptor agonist UK14304 are uncovered by pharmacological manipulation. As both neuropeptide Y (NPY) receptors and alpha2-adrenoceptors are negatively-coupled to adenylyl cyclase in this tissue, we determined whether NPY is also able to produce an enhanced contraction in the same tissue, under the same conditions. NPY (0.1 microM) produced a small contraction of porcine isolated ear arteries which was 5.1+/-0.8% of the response to 60 mM KCl (n = 14). An enhanced NPY response was uncovered if the tissue was pre-contracted with 0.1 microM U46619, and relaxed back to baseline with 1-2 microM forskolin before the addition of NPY (49.8+/-5.3%, n = 14). Forskolin (1 microM) stimulated cyclic AMP accumulation in porcine ear artery segments in the presence of 0.1 microM U46619 and 1 mM isobutylmethylxanthine (IBMX), NPY (0.1 microM) inhibited this response by 40%, but had no effect on basal levels of cyclic AMP. An enhanced response to 0.1 microM NPY was also obtained after pre-contraction with 0.1 microM U46619 and relaxation with either SNP (28.9+/-5.7%, n = 14), or dibutyryl cyclic AMP (21.2+/-4.6%, n = 14). This indicates that at least part of the enhanced response to NPY is independent of the agonist's ability to inhibit adenylyl cyclase. In conclusion, an enhanced contraction to NPY in the porcine isolated ear artery can be obtained by prior pharmacological manipulation. The enhanced responses are mediated through adenylyl cyclase-dependent and independent pathways similar to those reported for alpha2-adrenoceptors in this preparation.

Vasomotor Dysfunction Early after Exposure of Normal Rabbit Arteries to an Adenoviral Vector

We aimed to investigate whether infection of normal rabbit arteries with a recombinant adenovirus vector would result per se in alterations in contractile and endothelial functions. In one group of rabbits, right or left femoral and ear artery segments were injected in vivo with a replication-deficient adenoviral vector expressing a beta-galactosidase (beta-Gal) reporter gene (4 x 10(10) pfu/ml) to demonstrate efficient gene transfer. Contralateral arteries were injected with the same concentration of a recombinant adenoviral vector carrying no transgene (Ad.MLPnull). In another group of animals, Ad.MLPnull was injected into the lumen of femoral and ear artery segments. The contralateral arteries were used as controls with the injection of vehicle alone. Histochemical assessment of gene transfer using beta-Gal activity (group 1) or in vitro contractility and endothelial function (group 2) was performed 3 days after adenoviral infection. Gene transfer was efficient and reproducible in the endothelium and was associated with the presence of inflammatory cells in the media. In Ad.MLPnull-injected arteries, in vitro contractile response of femoral artery rings to either KCl 60 mM or phenylephrine (10 microM) was reduced to 10.5 +/- 2.3% (n = 14; p < 0.001) and 8.8 +/- 2.0% (n = 7; p < 0.001) of the control values, respectively. Furthermore, in arteries injected with Ad.MLPnull, the endothelium-dependent relaxation produced by acetylcholine (10 microM) was virtually abolished. Similarly, the relaxant effects of the alpha 2-adrenoreceptor agonist UK14304 (1 microM) or the Ca2+ ionophore A23187 (1 microM) were also abolished. By contrast, sodium nitroprusside (10 microM) was still able to relax adenovirus-infected arteries. We conclude that infection with a recombinant adenoviral vector can induce early severe vasomotor alterations in both contractile function and endothelium-mediated relaxation of normal rabbit arteries.

Intraarterial propofol is not directly toxic to vascular endothelium.

To determine if accidental intraarterial injection of propofol results in vascular damage, the effect of bolus administration of propofol on vascular smooth muscle and the endothelium was investigated using the isolated rabbit ear artery. Ear artery segments, removed from urethane anesthetized rabbits, were perfused with Krebs solution (1 ml.min-1) and pressurized to 60 mmHg before being constricted with extraluminal norepinephrine (1.8-4.2 x 10(-6) M). The external diameter of the vessel was measured by an array of light-dependent diodes. Functional responsiveness was determined by the degree of constriction to norepinephrine and the subsequent dilatation of the artery to intraluminal acetylcholine (2 x 10(-6) M) and glyceryl trinitrate (2 x 10(-6) M), and by the myogenic reactivity to a pressure increase from 60 to 100 mmHg. These responses were measured before and after perfusion with 1% propofol for 120 s. Administration of propofol did not result in any vasoactivity nor did it increase the sensitivity to norepinephrine. Vessels maintained their capacity to dilate to both agents, while the myogenic activity was unaffected. Histologic examination of the propofol exposed vessels showed no changes to smooth muscle structure, and the endothelial layer remained intact.

INTRA-ARTERIAL THIOPENTONE IS DIRECTLY TOXIC TO VASCULAR ENDOTHELIUM

Ear artery segments removed from urethane-anaesthetized rabbits were mounted, perfused with Krebs solution, and pressurized before constriction with extraluminal noradrenaline. Vessel diameter was measured using a diode array mounted above the artery. After the degree of dilatation in response to intraluminal acetylcholine and glyceryl trinitrate was measured, vessels were perfused for 120 s with solutions of either 2.5-10% thiopentone or isotonic sodium carbonate in saline of matched pH. Administration of thiopentone solutions in all concentrations resulted in destruction of endothelial cells. The dilatation of vessels to glyceryl trinitrate and their myogenic reactivity was unaltered. Isotonic solutions of sodium carbonate (pH 10.6) had no effect on either endothelial cell function or direct vasodilatation. These data show that administration of thiopentone removes arterial endothelial cells, while vascular smooth muscle function is essentially unaltered.

Myogenic response of isolated pressurized rabbit ear artery is independent of endothelium

Central ear artery segments, removed from urethan-anesthetized rabbits, were used to assess whether distention activation was dependent on intact and functional endothelium (ENDO). Changes in external arterial diameter were measured with light-dependent diode array mounted above the vessel. After constriction with norepinephrine, slow increases in pressure from 60 to 80 or 100 mmHg performed over 120 s (pressure ramps) failed to initiate distention, but initial diameter was maintained. Rapidly applied (500 ms) pressure increases (pressure jumps) across same pressure ranges gave rise to initial distention and a myogenic response resulting in a return to almost initial diameter while new pressure was maintained for 120 s. Myogenic activity was measured from extent of recovery of vessel to its initial diameter during maintenance of pressure jumps or ramps. Rapid jumps and slow ramps were performed in presence of an intact ENDO and again after ENDO was removed by passage of intraluminal gas. With pressure jumps 60 to 80 mmHg, degree of recovery in ENDO-denuded vessels was 91.9% compared with 89.5% in ENDO intact vessels. For pressure jumps 60 to 100 mmHg, extent of recovery was 87.4 and 89.6%. During application of pressure ramps of 20 or 40 mmHg, vessel diameter did not increase by greater than 5%. There are no significant differences in these data, and we conclude that myogenic response in rabbit ear artery is mediated independently of endothelial-derived factors, irrespective of whether myogenic activation is induced by pressure jumps or ramps.

Cryopreservation of innervation, endothelial and vascular smooth muscle function of a rabbit muscular and resistance artery.

The extent of preservation of endothelial, neurogenic and vascular function following frozen storage was studied using the rabbit central ear artery and a resistance artery branch. Fresh and frozen-stored artery segments were placed in tissue baths containing a physiological buffer solution and attached to a wire myograph apparatus for measurement of isometric force. Responses to cumulative additions of norepinephrine, histamine, acetylcholine and sodium nitroprusside were compared between fresh arteries and adjacent segments that had been stored frozen for 2-8 days at -70 degrees C. Responses to transmural nerve stimulation were measured in central ear artery segments that were fresh or stored frozen for 13-26 days. Large and small artery segments were frozen in ampules containing newborn calf serum and 1.8 M dimethylsulfoxide used as a cryoprotective agent. Following frozen storage, the vascular sensitivity of large and resistance arteries to norepinephrine and histamine were unaltered although maximal contractile responses were significantly reduced. Agonist affinity (KA values) to norepinephrine was also unchanged following frozen storage of large artery segments. Constrictor responses to varied frequencies of transmural nerve stimulation were similar between fresh and frozen-stored large arteries, suggesting that adrenergic nerve function is well preserved. In resistance arteries, vascular sensitivity and maximal relaxation to acetylcholine and sodium nitroprusside were unchanged. Similarly, relaxation of large arteries to sodium nitroprusside was well preserved. Although vascular sensitivity to acetylcholine was reduced in large arteries following frozen storage, much of the endothelial function was still preserved as indicated by only a 10% decrease in maximal relaxation.(ABSTRACT TRUNCATED AT 250 WORDS)

Myogenic adaptation of rabbit ear arteries to pulsatile internal pressures.

1. The effect of sinusoidal internal pressures on the constriction of in vitro, pressurized segments of ear arteries from rabbits has been examined. All arteries were constricted against a static transmural pressure of 60 mmHg to 35-60% of maximal, using extraluminal noradrenaline, before being exposed to the sinusoidal pressures. 2. There was a short period of adaptation when active arteries were first exposed to physiological pulsatile pressures. This adaptation had two components: a small, largely transient distension, lasting about 1 min, and sustained suppression of the distension produced by individual pressure pulses. 3. Constriction of adapted arteries was insensitive to physiological changes in pulse frequency (3-5 Hz), mean pulsatile pressure (60-120 mmHg), pulse amplitude (20-40 mmHg) and to alterations in pulse shape (sinusoidal, triangular and ramp). Over-all distension was restricted to 3.6 +/- 1.0% (S.D. of an observation) when the mean of a 3 Hz sinusoidal pressure of 30 mmHg amplitude was increased in steps from 75 to either 115 or 120 mmHg. 4. An initial distension was needed to initiate suppression of pressure pulse distension. Distension by individual pressure pulses, within a train of rectangular 60-90 mmHg pulses of 0.5 s duration, was maximally suppressed (85.6 +/- 1.3%; S.E. of mean, n = 9) at a pulse interval of 0.7 s. 5. Active ear arteries possess a myogenic mechanism capable of minimizing changes in constriction over the full physiological range of pulsatile internal pressures.

High-cholesterol diet induces altered responsiveness of rabbit arterial smooth muscle to noradrenaline.

The potential effect of feeding a high-cholesterol atherogenic diet (HCD) on in vitro responsiveness of rabbit arterial smooth muscle was investigated. The vasoconstrictor response produced by either exogenous or endogenous (field stimulation) noradrenaline was significantly depressed in isolated segments of ear artery after HCD as compared to regular diet fed animals. On the other hand no changes to high K+ could be observed in arterial segments after HCD as compared to regular diet fed rabbits. In addition, the alpha-adrenoceptor-mediated inotropic response and contraction of bladder smooth muscle showed only minor or no changes after HCD as compared to regular diet fed rabbits.

Proline and thymidine uptake in rabbit ear artery segments in vitro increased by chronic tangential load.

Indices of structural change were examined in blood vessel walls subjected to increased tangential load in a new, in vitro model system. Ring segments of rabbit ear artery were maintained in organ culture medium for times up to 9 days. Tangential load was chronically applied with small, intraluminal springs made of 0.010 in. diameter stainless steel wire. the applied load was considered to produce levels of circumferential wall tension corresponding to those induced by a range of levels of blood pressure. The indices of structural change examined were the uptake of radioactively-labelled proline and thymidine, which indicate protein synthesis and cell division respectively. Increased uptake of both proline and thymidine was noted in artery segments under elevated mechanical tension after a latency of 3 to 4 days. The degree of uptake was related to the degree of calculated wall tension elevation. The work indicated that cell division and protein synthesis can be induced in the blood vessel wall by increased wall tension alone, in vitro.

MODULATION OF NORADRENERGIC TRANSMISSION IN THE RABBIT EAR ARTERY BY DOPAMINE

1. The effects of dopamine on vasoconstrictor responses to field stimulation of sympathetic nerves and to exogenous noradrenaline were studied in the isolated ear artery of the rabbit. Responses to noradrenaline were unchanged at the start of the dopamine infusions but were enhanced as the infusions continued and also after cessation of the infustion. 2 Dopamine (0.5 muM) reduced the stimulation-induced efflux of tritium from segments of ear artery labelled with [3H]-noradrenaline. The reduction persisted during 65 min of dopamine infusion, after which time the vasoconstrictor responses had generally recovered to 93% of control level. On ceasing the infusion, the stimulation-induced efflux and the vasoconstrictor responses were enhanced. 3 Metoclopramide, haloperidol and ergometrine, each in a concentration of 0.2 muM, prevented the inhibitory effect of 0.5 muM dopamine on the stimulation-induced tritium release, but not the inhibitory effect of 0.5 muM noradrenaline. Phenoxybenzamine (0.2 and 1 muM) and phentolamine (1 muM) prevented the inhibitory effects of both noradrenaline and dopamine on the stimulation-induced efflux, and phentolamine (0.2 muM) prevented the inhibition of the stimulation-induced release by noradrenaline but only partially prevented the inhibitory effect of dopamine on the stimulation-induced efflux. 4 A possible role for dopamine in the modulation of noradrenergic transmission is suggested.