Biomedical subjects
U Pohl
Publications and source records attributed to U Pohl.
Elevation of plasma viscosity induces sustained NO-mediated dilation in the hamster cremaster microcirculation in vivo.
We studied whether a flow-independent increase of luminal wall shear stress (WSS) could dilate hamster arterioles in vivo and which endothelial mediators are potentially involved. To this end the plasma viscosity was elevated by exchanging blood for dextran-erythrocyte solution thereby augmenting WSS. Diameters of small and large arterioles as well as red blood cell velocities were measured before and after exchange of blood for solutions of identical haematocrit containing either high- (HMWD) or low-molecular weight dextran (LMWD). The potential role of endothelial autacoids was investigated by local application of the NO-synthase inhibitor NG-nitro-L-arginine (L-NNA), the inhibitor of cyclooxygenase, indomethacin (3 microM), or the K+-channel blocker, tetrabutylammonium (TBA, 0.1 mM) to assess the potential effects of EDHF. HMWD (n = 11 animals) increased plasma viscosity by 64 +/- 3% and dilated arterioles of all branching orders (A1-A4) significantly [by 24 +/- 3% (A1-A2) and 32 +/- 3% (A3-A4)]. This dilation compensated fully for the calculated initial increase of WSS. LMWD (n = 6) did not affect plasma viscosity or arteriolar diameters. Tissue treatment with L-NNA (30-300 microM, n = 12) substantially diminished the HMWD-induced dilation in small arterioles (A3-A4; to 13 +/- 3%; P<<0.05) and virtually abolished it in large ones (A1-A2). Consequently, the calculated WSS increased significantly in these arterioles (by 31 +/- 5%). TBA combined with L-NNA (n = 4) did not reduce further the remaining dilation. Indomethacin (n = 6) had no effect on HMWD-induced dilation. We conclude that an increase of WSS induces a mainly NO-mediated arteriolar dilation. This dilation occurs in all arteriolar branching orders and is of sufficient magnitude to compensate for the initial WSS-increase. Thus, any elevations of WSS fulfil the requirement for a signal to change diameter along the arteriolar tree in a coordinated manner. The fully compensating dilation which we observed indicates that WSS is a controlled variable. It does, however, raise questions as to its role as a continuous endothelial stimulus.
Indomethacin enhances endothelial NO release--evidence for a role of PGI2 in the autocrine control of calcium-dependent autacoid production.
OBJECTIVE: We studied whether NO or prostacyclin (PGI2), which are continuously released by endothelial cells, have autocrine/paracrine effects on the calcium-dependent autacoid production by modulating the intracellular Ca2+ concentration ([Ca2+]i). METHODS: Histamine(His)-induced [Ca2+]i increases (Fura 2-method) and NO-dependent cGMP increase were measured in human umbilical vein endothelial cell (HUVECs) before and after cyclooxygenase inhibition or application of cAMP- and cGMP-elevating drugs. RESULTS: 0.3 microM His increased endothelial [Ca2+]i from 77 +/- 2 nM to 418 +/- 59 nM. The His-induced [Ca2+]i increases were significantly attenuated following treatment with PGI2 (by 23%) and forskolin (by 33%), both increasing the cAMP release from HUVECs (by 49% and 66%). The His-induced [Ca2+]i increases were inhibited by the protein kinase A-activator cBIMPS (by 61%) which also abolished the His-induced PGI2 release. Conversely, inhibition of the PGI2 production with indomethacin significantly augmented the His-induced [Ca2+]i increases (by 32%), resulting in a significantly augmented NO production as indicated by an enhanced LNNA-sensitive cGMP increase in HUVECs. In contrast, neither increases of cGMP (basal 0.4 +/- 0.1 pmol/mg) elicited by 10 microM SNP (21 +/- 2 pmol/mg) or 10 microM C-type natriuretic peptide (CNP, 4.6 +/- 1.6 pmol/mg) nor its reduction by 30 microM LNNA had any effect on the His-induced [Ca2+]i increases. CONCLUSION: PGI2 attenuates agonist-induced [Ca2+]i increases by a cAMP-dependent mechanism, thereby modulating not only its own synthesis via a negative feedback but also that of NO. Consequently, reduced PGI2 levels result in an increased NO production. NO which does not cause a negative feedback control by cGMP might therefore compensate for the lack of PGI2.
Interaction of nitric oxide with myogenic and adrenergic vasoconstrictor processes in the control of microcirculatory blood flow.
Since the early observations of Carl Ludwig that the sympathetic nervous system exerts a tonic influence on resistance vessels, many additional factors have been identified which control vascular tone. The active modulator role of endothelium-derived nitric oxide (NO) in this process has emerged only in the last decade. The interaction of the local vasodilator, NO, with systemic alpha-adrenergic control of vascular tone as well as with pressure-induced myogenic vasoconstriction is briefly reviewed in the present paper. Cellular mechanisms of this interaction are discussed. In vitro and in vivo experiments indicate that the continuous basal release of NO potently attenuates alpha-adrenergic and myogenic tone in resistance vessels by several independent mechanisms and that endothelial impairment results in abnormal vasoconstriction.
Effects of LDL on intracellular free calcium and nitric oxide-dependent cGMP formation in porcine endothelial cells.
Nitric oxide (NO)-mediated, endothelium-dependent vasodilation is reduced in atherosclerotic arteries. A number of in vivo studies suggest that infusion of the substrate of NO synthase, L-arginine, partly counteracts this effect. We studied the potential inhibitory effects of native and of oxidized low density lipoproteins (n-LDL, ox-LDL) on NO-dependent cyclic guanidine monophosphate (cGMP) formation in porcine aortic endothelial cells and the role of extracellular L-arginine in counteracting this process. NO-dependent cGMP production in the cells (passage 1; preincubated in L-arginine-free medium for 24 h) was stimulated for 3 min with bradykinin (BK 1 nM) or the calcium-ionophore A23187 (100 nM) or by a 20 min incubation with L-arginine (L-Arg 0.1 mM, 20 min). The endothelium-independent NO-donor, sodium nitroprusside (SNP 1 microM) was used as control stimulus. Experiments were performed in the presence of LDL which was kept as much as possible under antioxidative conditions, further referred to as n-LDL (1 mg/ml), or LDL which was oxidized by incubation with copper (ox-LDL 0.1 mg/ml). The NG-nitro-L-arginine (L-NNA, inhibitor of NO-synthase) -sensitive intracellular cGMP concentration was taken as a measure of endothelial NO formation and determined by radioimmunoassay. BK-induced changes of intracellular free Ca2++ were measured immediately after washout of LDL using the FURA-2 method. n-LDL reduced the cGMP-levels in unstimulated cells as well as the cGMP increase in response to bradykinin (-10%) and the calcium-ionophore A23187 (-80%). The SNP-induced cGMP-increase was, however, not affected. L-arginine increased the intracellular cGMP concentration under both conditions by a similar amount, without affecting intracellular free calcium. Uptake of 3H-L-arginine was not significantly altered by n-LDL treatment. Ox-LDL significantly reduced SNP-induced cGMP-increases but did not alter bradykinin-induced cGMP increases. The BK-induced increase of intracellular free calcium was even enhanced after exposure of the cells to ox-LDL. L-arginine increased cGMP by a similar amount as in untreated cells. It is concluded that both n-LDL and ox-LDL can reduce the NO-dependent cGMP formation in cultured endothelial cells, albeit by different mechanisms. However, a limitation of the uptake or availability of extracellularly applied L-arginine does not appear to be a causal factor, at least not after 2 h exposure.
Synergistic action of vasodilators that increase cGMP and cAMP in the hamster cremaster microcirculation.
OBJECTIVE: Compounds such as endothelium derived nitric oxide (NO) and prostacyclin (prostaglandin I2) which increase cGMP and cAMP inhibit platelet activation in a synergistic manner. The aim of this study was to examine whether these compounds also interact synergistically in the control of smooth muscle tone. METHODS: Vascular diameters in the cremaster of 49 anaesthetised hamsters (465 arterioles) were studied during superfusion with compounds raising cAMP (isoprenaline and prostacyclin) and cGMP (sodium nitroprusside) alone or in combination. RESULTS: (1) The isoprenaline induced maximum dilator response was significantly attenuated, from 86.1(SEM 0.7)% to 37.1(0.2)%, after inhibition of NO-synthase with NG-nitro-L-arginine (L-NNA, 30 microM). Superfusion with sodium nitroprusside (30 nM, dilatation alone: 6.7%), which was used to substitute for endothelium derived NO, restored the attenuated isoprenaline response. The combined effects of isoprenaline and sodium nitroprusside were supra-additive. Virtually identical results were obtained when prostacyclin, another cAMP raising compound, was used instead of isoprenaline. The K+ channel opener cromakalim (100 nM) which acts cGMP independently was without effect on the prostacyclin induced dilator response. (2) The sodium nitroprusside induced maximum dilator response was attenuated from 80.9(0.25)% to 70.1(0.4)%, after indomethacin (3 microM) and restored by simultaneous application of prostacyclin (1 nM, dilatation alone: 1.4%) but not of cromakalim. Again, the combined effects were supra-additive, suggesting a synergistic action of these compounds. (3) Although indomethacin or L-NNA alone decreased the resting diameter by approximately 9.5%, the simultaneous application of both inhibitors failed to decrease the resting diameter further (10.0%, p = 0.97). CONCLUSIONS: Vasodilators increasing cGMP and cAMP act synergistically in vivo. Continuous release of NO and prostaglandins by the endothelium may therefore not only modulate the efficacy of such cyclic nucleotide increasing vasodilators but also interact synergistically in controlling basal vascular tone.
Attenuation of coronary autoregulation in the isolated rabbit heart by endothelium derived nitric oxide.
OBJECTIVE: The aim was to investigate the role of endothelium derived nitric oxide (EDNO/EDRF) in the control of coronary autoregulation. METHODS: In isolated saline perfused rabbit hearts coronary flow responses to stepwise increases in perfusion pressure were studied under control conditions, during maximum dilatation with sodium nitroprusside, and in the presence of the inhibitor of EDNO synthesis, NG-nitro-L-arginine (L-NNA), or the vasoconstrictors endothelin-1 and arginine vasopressin. RESULTS: At a constant perfusion pressure of 60 mm Hg, infusion of L-NNA (30 microM), but not D-NNA, reduced the coronary flow from 24.7(SEM 2) to 13.6(2.2) ml.min-1 and abolished flow increases induced by the EDRF stimulator acetylcholine. Under these conditions, pressure induced coronary flow increases were reduced (p < 0.05 compared to control) over the whole range of perfusion pressures studied (45 to 120 mm Hg). Arginine vasopressin [2(0.6) nM] and endothelin-1 [1.5(1) nM] induced similar reductions of coronary resting flow but the pressure induced flow increases were significantly greater than in the presence of L-NNA. Moreover, inhibition of EDRF synthesis reduced the peak reactive hyperaemia after a 30 s interruption of coronary flow from 47(2) to 32(2) ml.min-1. These changes occurred in spite of a decrease in the myocardial oxygen uptake from 5.1(0.6) to 3.4(0.5) ml.100 g-1.min-1 (p < 0.01) and a concomitant increase in the lactate release from 46(7) to 95(54) mumol.min.100 g-1 (p < 0.01), indicating myocardial ischaemia. CONCLUSIONS: EDNO attenuates coronary autoregulatory responses which, if unopposed, potentially impair a functionally adequate myocardial perfusion. It is suggested that the modulator role of EDNO is, at least in part, specific and most likely to be due to shear dependent alterations of EDNO release.
Endothelium-dependent phosphorylation of vasodilator-stimulated protein in platelets during coronary passage.
Compounds that elevate intraplatelet guanosine 3',5'-cyclic monophosphate (cGMP) or adenosine 3',5'-cyclic monophosphate (cAMP) stimulate the phosphorylation of a 46- to 56-kDa thrombocyte protein designated "vasodilator-stimulated protein" (VASP), which is most likely involved in the regulation of adhesion/aggregation. We investigated whether endothelium-derived relaxing factor (EDRF)/nitric oxide (NO) and prostaglandin I2 (PGI2) affected VASP phosphorylation in washed human platelets that were injected into the coronaries of saline-perfused rabbit hearts (n = 22) and collected immediately after passage. The endothelial stimulator acetylcholine (ACh; 1 microM) significantly increased the concentration of cGMP (indicating release of EDRF) and PGI2 in the coronary venous effluent, as well as the concentration of cGMP and cAMP in platelets. Phosphorylation state of VASP increased from 32.1 +/- 2.9 to 64.8 +/- 2.7%. Inhibition of EDRF/NO synthesis by NG-nitro-L-arginine (30 microM) completely abolished the ACh-induced cGMP increase, attenuated the cAMP-elevation without affecting PGI2, and caused a 20.5 +/- 5.8% decrease of the phosphorylation state of VASP. Indomethacin (30 microM) virtually abolished ACh-induced increases of PGI2, cAMP (but not cGMP), and phosphorylated VASP. These results indicate that both EDRF/NO and PGI2 contribute to the regulation of VASP phosphorylation in platelets collected after a single coronary passage. Their synergistic inhibitory effects on platelet function may thus be mediated by a common effect on target proteins like VASP as well as by a secondary increase in cAMP in response to cGMP-elevating compounds such as EDRF.
Low-dose natural interleukin-2 and recombinant interferon-gamma following autologous bone marrow grafts in pediatric patients with high-risk acute leukemia.
Twenty-two patients with high risk hematologic malignancies (13 c-ALL, two B-ALL/NHL, four T-ALL, two AML M2, one pre-pre B-ALL) entered a phase I/II trial with cyclic administration of low dose natural interleukin-2/recombinant interferon-gamma (nIL-2/rIFN-gamma) following autologous bone marrow transplantation (ABMT), in order to induce a cytotoxic antileukemic effect. Eighteen patients subsequently relapsed, corresponding to a Kaplan-Meier estimate of disease-free survival (DFS) of 18%. Compared with a historical group of autologous bone marrow recipients who have not received immunotherapy, there is no significant difference according to DFS. Immunophenotyping of peripheral lymphocytes at the onset and end of therapy cycles revealed the most significant mean increase among the NK cell population (262/microliters +/- 51 vs. 354/microliters +/- 36, p = 0.004). However, even CD3 positive T cells rose significantly (591/microliters vs. 689/microliters, p = 0.04). In vitro NK cell activity tested against the NK sensitive myeloid leukemic cell line K562, and LAK cell activity tested against the LAK sensitive Burkitt lymphoma cell line Raji, was only low. An additional in vitro stimulus with nIL2, however, led to a therapy-dependent increase of cytotoxicity which was significant against Raji cells (25% +/- 4 vs. 41% +/- 5, p = 0.0124) indicating that low dose nIL2/rIFN-gamma enhances precursors of potentially cytotoxic cells in vivo.
Interactions of hormones with the vascular endothelium. Effects on the control of vascular tone.
Due to their anatomical location vascular endothelial cells are an obvious target for hormones which are transported by the bloodstream. Studies on cultured endothelial cells, isolated vessels and the intact organism revealed the existence of multiple interactions between endothelial cells and circulating hormones. Not only are endothelial cells involved in the clearance of some specific circulating hormones, but they also form a tight barrier for other hormones thus preventing or attenuating their direct effects on vascular smooth muscle. Endothelial cells are also involved in the production of circulating angiotensin II by the angiotensin converting enzyme. Probably the most significant effect of hormones in vascular control is the ability of many of them to modulate the release of vasoactive autacoids such as nitric oxide, prostaglandins and endothelin-1. Aside from acute stimulating effects on autacoid production, some hormones, particularly steroids, exert chronic effects on vasoactive-factor gene expression. Apparently, in the control of vascular tone, interactions between circulating hormones and the endothelium play a major role. However, the functional significance of these interactions, especially in pathophysiologic conditions remains to be determined.
Adaptive changes of the vascular system in heart failure. Potential role of impaired endothelial vasomotor function.
An abnormality of myocardial cell function is the primary pathophysiologic basis of heart failure. However, complex alterations of the whole circulatory system are also pathophysiologically important. At an early stage these alterations represent adaptive mechanisms which compensate for the impaired cardiac output. Later on, however, they induce a vicious circle which leads to a further progression of heart failure. The circulatory alterations are due to changes of the activity of neuroendocrine, hormonal and local autocrineparacrine factors. The latter most likely include alterations of the endothelial vasomotor function. The potential significance of impaired endothelial NO-release in heart failure and present clinical evidence for such a mechanism are discussed.
Endothelium-derived nitric oxide in the control of tissue perfusion and oxygen supply: physiological and pathophysiological implications.
Since the development of specific NO-synthase inhibitors it has become possible to study the role of NO in the control of local blood flow and tissue oxygenation. Inhibition of NO-synthase induces hypertension and abnormal vasoconstriction, as well as tissue hypoxia and impaired adaptation of blood flow to increased tissue oxygen demands. These functional alterations are similar to those observed in a number of cardiovascular diseases. The present evidence that impaired endothelial function is a pathogenetic factor in the development of cardiovascular diseases is briefly reviewed.
[Methylene blue/light treatment of virus inactivated human plasma: production and clinical experience].
A photodynamic procedure to inactivate viruses in fresh plasma for therapeutical use is carried out by illuminating single units of plasma with visible light in the presence of the phenothiazine dye methylene blue. The blood bag systems of all common suppliers can be used for this purpose. Photodynamic treatment only moderately influences the activities of plasma proteins. One of the most sensitive parameters is the thrombin time which is prolonged, dependent on dye concentration and illumination time. Under the chosen conditions (1 microM methylene blue, 1 h illumination time, about 50,000 lx), the increase is approximately 25%. A drug monitoring study indicated that photodynamically treated fresh plasma is as well tolerated as conventional fresh-frozen plasma. Between February and the end of July 1992 about 31,000 units of the virus-inactivated product were distributed to clinics within Lower Saxony.
Mechanical deformation of vessel wall and shear stress determine the basal release of endothelium-derived relaxing factor in the intact rabbit coronary vascular bed.
We investigated the mechanisms that are responsible for the basal release of endothelium-derived relaxing factor (EDRF), which is likely to be identical with nitric oxide, in the intact coronary circulation. The increase in cGMP content of platelets passing through the coronary bed of the isolated rabbit heart was used as an index of EDRF release. Platelet cGMP content after passage through the heart under control conditions (flow rate of 20 ml/min) amounted to 0.50 +/- 0.10 pmol/mg protein. Inhibition of endothelial nitric oxide synthesis by 30 microM NG-nitro-L-arginine (L-NNA) reduced this amount by more than 60%. Increasing flow rate from 20 ml/min to 40 and 60 ml/min led to flow-dependent dilation as reflected by the subsequent drop in perfusion pressure after an initial rise. The flow-dependent dilation was associated with a significant increase in the normalized platelet cGMP content. L-NNA abolished completely both the flow-dependent dilation and the increase in platelet cGMP content. Increasing shear stress by a strong vasoconstriction (1 nM endothelin-1) at constant flow was also accompanied by a 2.5-fold increase in platelet cGMP content. To investigate whether mechanical forces applied to the vascular wall by the myocardial contraction cycle were also a stimulus for EDRF release, cardiac arrest was induced by a continuous infusion of mepivacaine (final concentration, 0.02%). Under these conditions, a decrease in platelet cGMP content comparable to that after nitric oxide synthesis inhibition was observed in the arrested heart.(ABSTRACT TRUNCATED AT 250 WORDS)
The development of neutralizing antibodies in a patient receiving subcutaneous recombinant and natural interleukin-2.
Systemic administration of interleukin-2 (IL-2) in humans may induce antibodies specific to IL-2. The case is reported of a patient with metastatic rectal carcinoma who was treated with long-term subcutaneous IL-2 and a combination of subcutaneous IL-2 and interferon-alpha 2b (IFN-alpha 2b). This patient developed nonneutralizing and neutralizing anti-IL-2 antibodies recognizing both the recombinant and natural cytokine. Detectable serum levels of neutralizing antibodies were accompanied by the inhibition of immune responsiveness to systemic IL-2 in vivo.
Impaired tissue perfusion after inhibition of endothelium-derived nitric oxide.
The effects of a blockade of the action or synthesis of endothelium-derived nitric oxide (EDRF) on vascular resistance and reactivity, platelet cGMP and tissue oxygenation were studied. Experiments were performed in isolated perfused rabbit hearts as well as in rabbit hindlimbs in vivo. In isolated hearts, perfusion with hemoglobin (6 microM) or NG-nitro-L-arginine (30 microM) significantly increased vascular resistance. The cGMP level in platelets passing through the coronary bed was found to be more than 50% lower than with intact EDRF production. EDRF inhibition also resulted in a reduced peak reactive hyperemia, an enhanced reactive vasoconstriction after a rapid increase in perfusion pressure (myogenic response), and in abolition of flow-dependent dilation of coronary resistance vessels. In rabbit hindlimbs, local blockade of EDRF-mediated dilations by gossypol resulted also in an increased vascular resistance and abolition of the increase in platelet cGMP induced by intraarterial infusion of acetylcholine. In addition, the oxygen uptake of the hindlimb (-46%) and the skeletal muscle pO2 were significantly reduced. It is concluded that continuously released EDRF has a functional role in maintaining adequate tissue perfusion and oxygen supply. Furthermore, the adaption of the vascular bed to rapid changes in flow and pressure is impaired after inhibition of EDRF.
NG-nitro-L-arginine antagonizes endothelium-dependent dilator responses by inhibiting endothelium-derived relaxing factor release in the isolated rabbit heart.
The effects of a recently described inhibitor of endothelial NO synthesis, NG-nitro-L-arginine (L-NNA), on the vasomotor responses to endothelium-dependent and independent vasodilators, and on the release of endothelium-derived relaxing factor (EDRF), were studied in the isolated saline-perfused rabbit heart. Infusion of L-NNA (30 microM) resulted in a 52 +/- 12% increase in basal coronary perfusion pressure. The vasomotor responses to 1 microM acetylcholine (ACh) and serotonin after L-NNA became biphasic, showing a small transient dilation followed by a pronounced vasoconstriction. In contrast, the dilation observed with sodium nitroprusside was not affected by L-NNA. None of the above-mentioned effects was elicited by the stereo-isomer D-NNA. Similarly, an increase in the basal coronary perfusion pressure by endothelin-1 (0.3 nM) to the same level as observed with L-NNA did not alter the vasomotor responses to ACh and sodium nitroprusside. The increase in cyclic GMP (cGMP) in platelets passing through the coronary vascular bed was used as an index of EDRF release. Platelet cGMP amounted to 0.50 +/- 0.10 pmol/mg protein after passage through the coronary bed of the unstimulated heart. When platelets were injected during an ACh infusion (1 microM), a 2.7 fold increase in cGMP was observed (P less than 0.01). After a 30-min infusion with L-NNA, the cGMP content of platelets passing through the unstimulated heart was reduced by 62%. Likewise, the ACh-induced increase in platelet cGMP was totally blocked. These results show that L-NNA inhibits EDRF release, and is thus a potent and selective inhibitor of EDRF-mediated dilation in the isolated rabbit heart.
Effects of vasoactive agonists on the membrane potential of cultured bovine aortic and guinea-pig coronary endothelium.
1. The effects of bradykinin, ATP, adenosine, histamine and thrombin on the membrane potential of confluent monolayers of cultured bovine aortic endothelial cells (BAECs) and guinea-pig coronary endothelial cells (GCECs) were studied at 37 degrees C using the whole-cell mode of the patch-clamp technique. 2. The amplitude histogram of the resting potentials of BAEC monolayers showed a bimodal distribution with one peak around -25 mV and another peak around -85 mV. Transitions from one potential level to the other were observed. The bistable membrane potential can be explained by an N-shaped current-voltage relation of the endothelial cell membrane. 3. When BAECs with a low resting potential (-10 to -30 mV) were superfused with maximally effective concentrations of ATP (2-10 microM) an initial hyperpolarization of -80 to -90 mV was observed which decayed to a plateau of about -60 mV within 1 min. When ATP was removed after 2-3 min the membrane potential returned to control level within 1 min. This was followed by a second hyperpolarization of 10-20 mV, which decayed within 15 min. 4. In the absence of extracellular calcium, ATP produced only a brief transient hyperpolarization in aortic endothelium. The plateau and the secondary hyperpolarization were abolished. These findings are consistent with the idea that the changes in membrane potential reflect changes in intracellular free Ca2+ and that the initial peak is due to release of Ca2+ from intracellular stores, whereas the plateau and the secondary hyperpolarization depend on transmembrane Ca2+ influx. 5. Bradykinin evoked potential changes similar to ATP in BAECs, except that the secondary hyperpolarization during wash-out was absent. When the membrane potential was more negative than -80 mV, ATP and bradykinin induced only a small initial hyperpolarization followed by a depolarization of up to 20 mV. 6. In aortic endothelium, ADP (10 microM) evoked a much smaller response than ATP. Adenosine (10 microM), thrombin (2 units/ml), acetylcholine (10 microM) and histamine (10 microM) had only a very small effect on the membrane potential, if any. 7. The amplitude histogram of the membrane potential of GCECs showed only one peak around -35 mV. In coronary endothelium, application of bradykinin, ATP, histamine, thrombin, acetylcholine and adenosine all evoked a transient hyperpolarization of 10-40 mV lasting 1 min or less, which then turned into a depolarization. 8. The K+ channel openers cromakalim (BRL 34915) and lemakalim (BRL 38227) did not affect the membrane potential of GCECs or BAECs.(ABSTRACT TRUNCATED AT 400 WORDS)