Endothelial cell signaling during conducted vasomotor responses

2003 ◽  
Vol 285 (1) ◽  
pp. H119-H126 ◽  
Author(s):  
Kim A. Dora ◽  
Jun Xia ◽  
Brian R. Duling
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Membrane Potential ◽  
Endothelial Cell ◽  
Electrical Coupling ◽  
Cheek Pouch ◽  
Application Site ◽  
Hamster Cheek Pouch ◽  
Vasomotor Responses ◽  
Site Of Stimulation

ACh and KCl stimulate vasomotor responses that spread rapidly and bidirectionally along arteriole walls, most likely via spread of electric current or Ca2+ through gap junctions. We examined these possibilities with isolated, cannulated, and perfused hamster cheek pouch arterioles (50- to 80-μm resting diameter). After intraluminal loading of 2 μM fluo 3 to measure Ca2+ or 1 μM di-8-ANEPPS to measure membrane potential, photometric techniques were used to selectively measure changes in intracellular Ca2+ concentration ([Ca2+]i) or membrane potential in endothelial cells. Activation of the endothelium by micropipette application of ACh (10-4 M, 1.0-s pulse) to a short segment of arteriole (100–200 μm) increased endothelial cell [Ca2+]i and caused hyperpolarization at the site of stimulation. This response was followed rapidly by vasodilation of the entire arteriole (∼2-mm length). Change in membrane potential always preceded dilation, both at the site of stimulation and at distant sites along the arteriole. In contrast, an increase in endothelial cell [Ca2+]i was observed only at the application site. Micropipette application of KCl, which can depolarize both smooth muscle and endothelial cells (250 mM, 2.5-s pulse), also caused a rapid, spreading response consisting of depolarization followed by vasoconstriction. With KCl stimulation, in addition to changes in membrane potential, increases in endothelial cell [Ca2+]i were observed at distant sites not directly exposed to KCl. The rapid longitudinal spread of both hyperpolarizing and depolarizing responses support electrical coupling as the mode of signal transmission along the arteriolar length. In addition, the relatively short distance between heterologous cell types enables the superimposed radial Ca2+ signaling between smooth muscle and endothelial cells to modulate vasomotor responses.

Role of EDHF in conduction of vasodilation along hamster cheek pouch arterioles in vivo

2000 ◽  
Vol 278 (6) ◽  
pp. H1832-H1839 ◽  
Author(s):  
Donald G. Welsh ◽  
Steven S. Segal
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Membrane Potential ◽  
Smooth Muscle Cells ◽  
Cheek Pouch ◽  
Hamster Cheek Pouch ◽  
Cytochrome P 450 ◽  
Arteriolar Diameter

We tested whether local and conducted responses to ACh depend on factors released from endothelial cells (EC) in cheek pouch arterioles of anesthetized hamsters. ACh was delivered from a micropipette (1 s, 500 nA), while arteriolar diameter (rest, ∼40 μm) was monitored at the site of application (local) and at 520 and 1,040 μm upstream (conducted). Under control conditions, ACh elicited local (22–65 μm) and conducted (14–44 μm) vasodilation. Indomethacin (10 μM) had no effect, whereas N ω-nitro-l-arginine (100 μM) reduced local and conducted vasodilation by 5–8% ( P < 0.05). Miconazole (10 μM) or 17-octadecynoic acid (17-ODYA; 10 μM) diminished local vasodilation by 15–20% and conducted responses by 50–70% ( P < 0.05), suggesting a role for cytochrome P-450 (CYP) metabolites in arteriolar responses to ACh. Membrane potential ( E m) was recorded in smooth muscle cells (SMC) and in EC identified with dye labeling. At rest (control E m, typically −30 mV), ACh evoked local (15–32 mV) and conducted (6–31 mV) hyperpolarizations in SMC and EC. Miconazole inhibited SMC and EC hyperpolarization, whereas 17-ODYA inhibited hyperpolarization of SMC but not of EC. Findings indicate that ACh-induced release of CYP metabolites from arteriolar EC evoke SMC hyperpolarization that contributes substantively to conducted vasodilation.

Capillaries demonstrate changes in membrane potential in response to pharmacological stimuli

1998 ◽  
Vol 274 (1) ◽  
pp. H60-H65 ◽  
Author(s):  
Eugene D. McGahren ◽  
James M. Beach ◽  
Brian R. Duling
Keyword(s):  
Endothelial Cells ◽  
Membrane Potential ◽  
Fluorescence Emission ◽  
Cheek Pouch ◽  
Surrounding Tissue ◽  
Hamster Cheek Pouch ◽  
Capillary Membrane ◽  
Resistance Vessels ◽  
Capillary Endothelial Cells

It has been proposed that capillaries can detect changes in tissue metabolites and generate signals that are communicated upstream to resistance vessels. The mechanism for this communication may involve changes in capillary endothelial cell membrane potentials which are then conducted to upstream arterioles. We have tested the capacity of capillary endothelial cells in vivo to respond to pharmacological stimuli. In a hamster cheek pouch preparation, capillary endothelial cells were labeled with the voltage-sensitive dye di-8-ANEPPS. Fluorescence from capillary segments (75–150 μm long) was excited at 475 nm and recorded at 560 and 620 nm with a dual-wavelength photomultiplier system. KCl was applied using pressure injection, and acetylcholine (ACh) and phenylephrine (PE) were applied iontophoretically to these capillaries. Changes in the ratio of the fluorescence emission at two emission wavelengths were used to estimate changes in the capillary endothelial membrane potential. Application of KCl resulted in depolarization, whereas application of the vehicle did not. Application of ACh and PE resulted in hyperpolarization and depolarization, respectively. The capillary responses could be blocked by including a receptor antagonist (atropine or prazosin, respectively) in the superfusate. We conclude that the capillary membrane potential is capable of responding to pharmacological stimuli. We hypothesize that capillaries can respond to changes in the milieu of surrounding tissue via changes in endothelial membrane potential.

scholarly journals Endothelial coordination of cerebral vasomotion via myoendothelial gap junctions containing connexins 37 and 40

2006 ◽  
Vol 291 (5) ◽  
pp. H2047-H2056 ◽  
Author(s):  
Rebecca E. Haddock ◽  
T. Hilton Grayson ◽  
Therese D. Brackenbury ◽  
Kate R. Meaney ◽  
Craig B. Neylon ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Endothelial Cell ◽  
Gap Junctions ◽  
Basilar Artery ◽  
Electrical Coupling ◽  
Potential Oscillations ◽  
Membrane Potential Oscillations ◽  
Rhythmical Contractions ◽  
Conventional Electron Microscopy

Control of cerebral vasculature differs from that of systemic vessels outside the blood-brain barrier. The hypothesis that the endothelium modulates vasomotion via direct myoendothelial coupling was investigated in a small vessel of the cerebral circulation. In the primary branch of the rat basilar artery, membrane potential, diameter, and calcium dynamics associated with vasomotion were examined using selective inhibitors of endothelial function in intact and endothelium-denuded arteries. Vessel anatomy, protein, and mRNA expression were studied using conventional electron microscopy high-resolution ultrastructural and confocal immunohistochemistry and quantitative PCR. Membrane potential oscillations were present in both endothelial cells and smooth muscle cells (SMCs), and these preceded rhythmical contractions during which adjacent SMC intracellular calcium concentration ([Ca2+]i) waves were synchronized. Endothelium removal abolished vasomotion and desynchronized adjacent smooth muscle cell [Ca2+]i waves. NG-nitro-l-arginine methyl ester (10 μM) did not mimic this effect, and dibutyryl cGMP (300 μM) failed to resynchronize [Ca2+]i waves in endothelium-denuded arteries. Combined charybdotoxin and apamin abolished vasomotion and depolarized and constricted vessels, even in absence of endothelium. Separately, 37,43Gap27 and 40Gap27 abolished vasomotion. Extensive myoendothelial gap junctions (3 per endothelial cell) composed of connexins 37 and 40 connected the endothelial cell and SMC layers. Synchronized vasomotion in rat basilar artery is endothelium dependent, with [Ca2+]i waves generated within SMCs being coordinated by electrical coupling via myoendothelial gap junctions.

Endothelial and smooth muscle cells hyperpolarized by bradykinin are not dye coupled

1990 ◽  
Vol 258 (3) ◽  
pp. H836-H841 ◽  
Author(s):  
J. L. Beny
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Endothelial Cell ◽  
Smooth Muscle Cells ◽  
Lucifer Yellow ◽  
Electrical Response ◽  
Electrical Coupling ◽  
Smooth Muscles ◽  
Muscle Cells ◽  
Electrical Signal

The membrane potential of endothelial and neighboring (0.1 mm) smooth muscle cells of pig coronary arteries were simultaneously recorded with two microelectrodes. The membrane potential of endothelial cells was -40 +/- 4 mV (n = 9). In these cells bradykinin (250 nM), an endothelium-dependent vasodilator, evoked a transient hyperpolarization (14 +/- 2 mV, n = 9) resembling those already observed in smooth muscles. The similarity between the electrical signal of pre- and postmyoendothelial junctions suggested an electrical coupling between endothelial and smooth muscle cells. However, the injection of the fluorescent dye Lucifer yellow in the recorded cell proved that the cell was endothelial, and in addition, the injection demonstrated the absence of dye coupling between endothelial and smooth muscle cells. Moreover the injection of electrical pulses (0.05-3.5 nA) in the endothelial cell never evoked any electrical response in the smooth muscle. By contrast, the smooth muscle cells were electrically coupled-together. These results do not support the idea that the endothelial cell hyperpolarization caused by bradykinin is transmitted to smooth muscle cells by electrotonic spreading.

Electromechanical coupling and the conducted vasomotor response

1995 ◽  
Vol 269 (6) ◽  
pp. H2022-H2030 ◽  
Author(s):  
J. Xia ◽  
B. R. Duling
Keyword(s):  
Membrane Potential ◽  
Electromechanical Coupling ◽  
Mechanical Response ◽  
Electrical Response ◽  
Threshold Level ◽  
Cheek Pouch ◽  
Resting Membrane Potential ◽  
Vasomotor Response ◽  
Hamster Cheek Pouch ◽  
Vasomotor Responses

Conducted vasomotor responses are viewed as one mechanism that functionally integrates the microvasculature. It is hypothesized that the conducted vasomotor response is the result of an electrical current and its passive electrotonic spread along the length of a microvessel. We tested this hypothesis in isolated, unpressurized arterioles from the hamster cheek pouch using conventional intracellular membrane potential recording techniques. The mean resting membrane potential (RMP) was -67 mV. KCl and phenylephrine (PE) pulse-stimulation applied through micropipettes could both induce transient depolarizations and vasoconstrictions at the site of stimulation (local) and at conducted (560 microns) sites. It was noted, however, that the conducted vasomotor response could not be induced until the conducted electrical response exceeded a threshold of -45 mV for a minimum amount of time. The relationship between the amplitude of constriction and the amplitude-time area of depolarization above -45 mV was the same for local and conducted KCl and for conducted PE but was significantly different from that for local PE. Nifedipine greatly reduced the local and conducted mechanical but not electrical responses. Our results indicate that the conducted vasomotor responses are the result of the generation and subsequent conduction of electrical signals along the vessel but that the corresponding mechanical response occurs only when the electrical response exceeds a threshold level.

Participation of intracellular Ca2+ stores in arteriolar conducted responses

2003 ◽  
Vol 285 (1) ◽  
pp. H65-H73 ◽  
Author(s):  
Yasuaki Yashiro ◽  
Brian R. Duling
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Pressure Pulse ◽  
Cell Types ◽  
Vasomotor Responses ◽  
Local Site ◽  
Intracellular Ca ◽  
Baseline Diameter ◽  
Site Of Stimulation ◽  
Conducted Response

We examined the role played by intracellular Ca2+ stores in conducted vasomotor responses induced by phenylephrine (PE) in isolated hamster cremasteric arterioles. When applied briefly (∼1 s) to isolated, cannulated arterioles by using pressure-pulse ejection from a micropipette, PE produced a strong local vasoconstriction and a very small biphasic conducted response (a small constriction followed by a dilation) that propagated several hundred micrometers along the vessel length. The conducted vasomotion was associated with a monophasic elevation of the endothelial cell intracellular Ca2+ concentration ([Ca2+]i) at the site of stimulation, as measured with the Ca2+ indicator fura 2. The Ca2+ pump inhibitor thapsigargin was used to limit filling of Ca2+ stores in smooth muscle and endothelial cells. Thapsigargin reduced baseline diameter and elicited a strong dilator component at the local site while enhancing both the constrictor and dilator components of the PE-induced conducted response. The enhanced conducted constrictor component induced by thapsigargin was mimicked by extraluminal application of tetraethylammonium or charybdotoxin but not by iberiotoxin, apamin, glibenclamide, barium, or 4-aminopirydine. Thapsigargin increased the estimated basal endothelial cell [Ca2+]i by ∼60 nM and converted the PE-induced change in [Ca2+]i from monotonic to biphasic with a late elevation of [Ca2+]i above baseline that coincided with the increased dilatory component of the conducted response. Luminal application of charybdotoxin plus apamin significantly reduced the dilatory component of the conducted response. These results indicate that intracellular Ca2+ stores play a dynamic role in regulating conducted vasomotor responses apparently through modulation of KCa channels in both cell types.

Length-tension relationship of vascular smooth muscle in single arterioles

1989 ◽  
Vol 256 (3) ◽  
pp. H630-H640 ◽  
Author(s):  
M. J. Davis ◽  
R. W. Gore
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Wall Stress ◽  
Physiological Saline ◽  
Intraluminal Pressure ◽  
Cheek Pouch ◽  
Hamster Cheek Pouch ◽  
Longitudinal Response ◽  
Physiological Saline Solution ◽  
Relationship Of

Longitudinal response gradients in the microcirculation may in part be explained in terms of the length-tension relationship of vascular smooth muscle at different points along the vascular tree. To test this hypothesis, four branching orders of arterial vessels (20-80 microns ID) were dissected from the hamster cheek pouch and cannulated with concentric micropipettes. Intraluminal pressure was monitored with a servo-null micropipette, and arteriolar dimensions were measured using a videomicrometer. All arterioles developed spontaneous tone in physiological saline solution. Pressure-diameter curves were recorded for maximally activated vessels and for passive vessels. Maximal active wall tension varied nearly threefold, but maximal active medial wall stress (approximately 4 x 10(6) dyn/cm2) varied only approximately 20% between the different vessel orders. These data support the concept that smooth muscle cells from vessels of different sizes are mechanically similar but do not completely explain the longitudinal response gradients reported in the cheek pouch microcirculation. An analysis of the effect of arteriolar wall buckling suggests that the luminal folds that develop at short vessel radii may broaden the peak of the active stress-length curve and extend the pressure range over which arterioles are most sensitive to physical and chemical stimuli.

Electrical activation of endothelium evokes vasodilation and hyperpolarization along hamster feed arteries

2001 ◽  
Vol 280 (1) ◽  
pp. H160-H167 ◽  
Author(s):  
Geoffrey G. Emerson ◽  
Steven S. Segal
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Endothelial Cell ◽  
Smooth Muscle Cells ◽  
Cell Damage ◽  
Muscle Cells ◽  
Sympathetic Nerves ◽  
Retractor Muscle ◽  
Receptor Interactions ◽  
Feed Arteries

Endothelial cells are considered electrically unexcitable. However, endothelium-dependent vasodilators (e.g., acetylcholine) often evoke hyperpolarization. We hypothesized that electrical stimulation of endothelial cells could evoke hyperpolarization and vasodilation. Feed artery segments (resting diameter: 63 ± 1 μm; length 3–4 mm) of the hamster retractor muscle were isolated and pressurized to 75 mmHg, and focal stimulation was performed via microelectrodes positioned across one end of the vessel. Stimulation at 16 Hz (30–50 V, 1-ms pulses, 5 s) evoked constriction (−20 ± 2 μm) that spread along the entire vessel via perivascular sympathetic nerves, as shown by inhibition with tetrodotoxin, ω-conotoxin, or phentolamine. In contrast, stimulation with direct current (30 V, 5 s) evoked vasodilation (16 ± 2 μm) and hyperpolarization (11 ± 1 mV) of endothelial and smooth muscle cells that conducted along the entire vessel. Conducted responses were insensitive to preceding treatments, atropine, or N ω-nitro-l-arginine, yet were abolished by endothelial cell damage (with air). Injection of negative current (≤1.6 nA) into a single endothelial cell reproduced vasodilator responses along the entire vessel. We conclude that, independent of ligand-receptor interactions, endothelial cell hyperpolarization evokes vasodilation that is readily conducted along the vessel wall. Moreover, electrical events originating within a single endothelial cell can drive the relaxation of smooth muscle cells throughout the entire vessel.

Ca2+ sensitivity of isolated arterioles from the hamster cheek pouch

1991 ◽  
Vol 260 (2) ◽  
pp. H355-H361 ◽  
Author(s):  
M. R. Hynes ◽  
B. R. Duling
Keyword(s):  
Smooth Muscle ◽  
Muscle Layer ◽  
Cheek Pouch ◽  
Maximum Diameter ◽  
Maximal Response ◽  
Bathing Solution ◽  
Hamster Cheek Pouch ◽  
Mean Time ◽  
Concentration Response Curve

When isolated from the hamster cheek pouch, cannulated, and perfused, 60- to 90-microns arterioles spontaneously contracted to 67 +/- 4% of maximum diameter. Vessel sensitivity to variations in extracellular Ca2+ was then evaluated. Tone, regardless of its source, was highly dependent on the concentration of Ca2+ in the bathing solution. The magnitude of responses to changing Ca2+ depended upon which vessel surface (luminal or abluminal) the change was made. For K(+)-induced tone the Ca2+ concentration-response curve was right shifted 60-fold for luminal vs. abluminal changes. These results suggest that restricted diffusion of Ca2+ from lumen to smooth muscle dramatically reduces smooth muscle Ca2+ concentration and that under standard in vitro conditions the smooth muscle layer is effectively isolated from luminal contents. Both the cytosolic and stored Ca2+ in these microvessels were dependent on the Ca2+ concentration in the bathing solution. Abrupt removal of Ca2+ from bath produced a rapid maximal dilation with a mean time to half-maximal response (t1/2 max) of 14 +/- 4 s. Ca2+ replacement induced a return to the previous level of tone with a mean t1/2 max of 8 +/- 3 s. The magnitude of transient responses to caffeine (10 mM) was inversely related to the time of exposure to zero Ca2+ with a rapid decay in magnitude (t1/2 max = 2.7 +/- 0.8 min). These data suggest that the smooth muscle cells of arterioles have a particularly rapid transmembrane Ca2+ flux that is tightly controlled by an intracellular regulatory mechanism, which may explain the generally increased dependence of smaller vessels on extracellular Ca2+.

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