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D R Harder

Publications and source records attributed to D R Harder.

At least 109 records · Page 6Linked to original sources
PubMed1982

Effect of H+ and elevated PCO2 on membrane electrical properties of rat cerebral arteries.

Some membrane electrical properties of muscle cells from the middle cerebral artery of the rat were recorded with intracellular microelectrodes. The resting membrane potential (Em) of this preparation was -63 mV. Reduction of extracellular pH to 7.0 in the face of a constant PCO2 of 40 mm Hg had no significant effect on Em. Similarly the slope of the steady-state voltage/current curves was not different at pH 7.0 compared to control at pH 7.4. In marked contrast, when PCO2 was elevated to around 60 to 70 mm Hg there was a rapid hyperpolarization and reduction in the slope of the voltage current curve suggesting an increased conductance for one or more ionic species. In addition elevation of PCO2 increased the slope of the Em vs. log[K]0 curve from 46 mV/decade to 59 mV/decade which is in good agreement with a Nernstian potential for a K+ selective membrane. These data suggest that while the smooth muscle cells of rat cerebral arteries are relatively insensitive to a small reduction in extracellular pH; reduction of intracellular pH by elevating PCO2 induces hyperpolarization by increasing K+ conductance (gk). However, it is not clear from these experiments if the PCO2 effects are mediated entirely by changes in pH or if there is a direct membrane action of CO2.

Animals↗
PubMed1982

Sympathetic supraspinal control of venous membrane potential in spontaneous hypertension in vivo.

To clarify the mechanisms controlling mesenteric venous membrane potential (Em) in spontaneously hypertensive rats (SHR) and Wistar-Kyoto normotensive rats (WKY), Em was measured in vivo using flexibly mounted glass microelectrodes. Venous diameters were also measured. Ems under control conditions were significantly less in SHR. alpha-Adrenergic blockade with phenoxybenzamine (PBZ) significantly dilated and hyperpolarized SHR small veins but failed to alter the membrane potential of WKY small veins. After blockade with PBZ, membrane potentials were similar in both strains. Propranolol suffusion failed to alter the SHR membrane potential but depolarized WKY small veins. During blockade with PBZ, propranolol depolarized venous membranes equally in both strains. Membrane potentials after spinal section were similar in both strains and equal to the Ems during total adrenergic blockade. The data indicate that, in SHR, the relatively depolarized Em derives from altered alpha-adrenergic input. In SHR, alpha-adrenergic input appears to exert its depolarizing effect, at least in party, by interfering with the expression of endogenous, beta-adrenergic hyperpolarization. The beta-adrenergic influence, like the alpha-adrenergic input in SHR, is dependent on intact supraspinal pathways.

Animals↗
PubMed1981

Membrane activation of smooth muscle from rabbit basilar artery by dopamine.

Intracellular membrane potential (EM) and force development were measured in rabbit basilar artery to help elucidate the mechanism of action of dopamine in this preparation. There was a strong correlation between membrane depolarization and contraction (r = 0.95) between 3 X 10-7 M to 10-4 M dopamine. When the vascular muscle cells were depolarized by elevating [K]0 there was a Em dependent decrease in force development in response to dopamine. Significant reduction of dopamine stimulated force development was observed when the vessel was depolarized by 5-6 mV by excess extracellular K+ and 90% inhibition was seen when the artery was depolarized to -20mV. When Ca++ influx was blocked, dopamine no longer induced force development. Such findings suggest that dopamine contracts rabbit basilar artery by a mechanism involving membrane depolarization. This process may involve an influx of extracellular Ca++ through voltage sensitive channels.

Animals↗
PubMed1981

Norepinephrine effect on in situ venous membrane potential in spontaneously hypertensive rats.

Comparative in situ (innervation and circulation intact) and in vitro measurements of transmembrane potential (Em) were made in vascular smooth muscle cells (VSM) of small (300-500 micrometers) veins of an externalized intestinal mesenteric loop in 13- to 15-wk-old anesthetized spontaneously hypertensive rats (SHR) and Wistar-Kyoto normotensive controls (WKY). During suffusion with physiological salt solution (PSS), the mean in situ Em in SHR was significantly lower (-34 +/- 0.8 mV) than in WKY (-49 +/- 1.3 mV). In situ neural blockade with 1 microgram/ml tetrodotoxin (TTX) in PSS hyperpolarized venous Em in SHR (-45 +/- 1.7 mV) but not in WKY. In vitro Em in SHR (-51 +/- 1.0 mV) and WKY (-54 +/- 1.1 mV), though significantly elevated above respective in situ values, did not differ significantly from each other and were not altered by TTX. Increasing norepinephrine (NE) concentrations in the PSS gradedly depolarized the venous VSM of WKY in situ to a plateau Em of -32 +/- 0.9 mV at 6 microM (1 micrograms/ml) NE but had no significant depolarizing effect on the less-polarized venous VSM of SHR. However, after addition of 1 micrograms/ml TTX to the PSS, the in situ Em in SHR followed a depolarizing NE dose-response curve similar to that observed in WKY (with or without TTX). These results support the hypothesis that the neurogenic vasoconstrictor component of VSM tone is significantly elevated in mesenteric veins of the SHR model of essential hypertension.

Animals↗
PubMed1981

Membrane electrical mechanism of basilar artery constriction and pial artery dilation by norepinephrine.

To study the mechanism by which norepinephrine acts on vascular muscle cell membrane, we recorded membrane potential with intracellular microelectrodes in isolated cat basilar and pial arteries. On addition of norepinephrine concentrations less than 1 microM, pial arteries hyperpolarized and relaxed while basilar arteries depolarized and contracted. Relaxation and hyperpolarization of the pial arteries occurred without the need for addition of any other drug, which indicates the relaxation of spontaneous tone. The relaxation and hyperpolarization could be completely blocked by addition of propranolol before exposure to norepinephrine. The depolarization and contraction of both basilar and pial arteries was blocked by the previous exposure to phentolamine. Electrical spikes were not found spontaneously, but could be induced in both arteries by tetraethylammonium and subsequent addition of norepinephrine, blockable by phentolamine. We conclude that membrane property differences between basilar and pial arteries result in qualitatively different effects of norepinephrine.

Animals↗
PubMed1980

Membrane potential measurements during rat liver regeneration.

Membrane potential was measured in perfused rat liver and was shown to increase from -33 +/- 1.0 mV in livers from normal rats to -50 +/- 1.1 mV in livers from rats 12 hr after partial hepatectomy. The hyperpolarization of the membrane in regenerating liver was no longer evident after perfusion with 1 mM ouabain for 5 min. Ouabain had a small (4 mV) depolarizing effect on membrane potential in normal liver. The potential measured in normal and regenerating liver decreased as a function of the external potassium concentration above 5 mM; however, the potential was more electronegative in regenerating liver compared to normal liver at all values of external potassium concentration, and the differences in potential between the two kinds of cells did not decrease at higher concentrations of external potassium. Thus, a plot of membrane potential vs external potassium concentration resulted in approximately parallel curves for the two different cell types. We conclude that hyperpolarization of the liver cell membrane is an early event during rat liver regeneration and results from an electrogenic Na-K pump.

Animals↗
PubMed1980

Transmembrane potential and amino acid transport in rat hepatocytes in primary monolayer culture.

Transmembrane potential (Em) and alpha-aminoisobutyric acid (AIB) transport were measured in primary monolayer cultures of rat hepatocytes obtained from unoperated control rats and from rats 12 hr following partial hepatectomy. Measurements were performed 20-24 hr after plating the cells. The capacity of both kinds of cells to concentrate AIB depended upon extracellular sodium: however, the steady-state accumulation in regenerating cells was twice that of control cells. Transmembrane potentials, recorded with glass microelectrodes, were -13 +/- 0.6 mV and -27 +/- 1.6 mV in control and regenerating cells, respectively. Ouabain (1 mM) depolarized regenerating cell to -18 +/- 1.0 mV, but it had no effect on control cells. The initial rates of 1 mM AIB transport into control and regenerating cells were 1.2 +/- 0.1 and 3.1 +/- 0.1 nanomoles/mg protein x 4 min, respectively. Ouabain (1 mM) reduced the initial rate of AIB transport into regenerating cells to 2.7 +/- 0.1 nanomoles/mg protein x 4 min, but it had no effect on AIB transport into control cells. Glucagon (10(-7) M) added to control cells 12 hr before measurements hyperpolarized Em to -31 +/- 1.3 mV and increased AIB transport rate to 3.1 nanomoles/mg protein x 4 min. The results suggest a relationship between increases in Em and increases in AIB transport in rat hepatocytes. An electrogenic Na-K pump may be involved in both of these events.

Aminoisobutyric Acids↗
PubMed1980

Active vascular smooth muscle tone and venous membrane potentials during hemorrhage.

To clarify mechanisms leading to failure of compensatory vascular tone in splanchnic blood vessels during prolonged hypotensive stress, anesthetized rats were maintained at a constant mean arterial pressure of 35 mmHg by hemorrhage into an external reservoir until 40% autoinfusion of maximum bled volume. In vivo intracellular membrane potentials (Em) of small intestinal mesenteric veins (300--500 micrometers) were measured before and during the compensatory (bleedout) and decompensatory (autinfusion) phases of the hypotensive period to assess the state of vascular smooth muscle (VSM) excitation. During the compensatory phase, Em decreased from -41 +/- 1 mV (prehemorrhage) to -31 +/- 2 mV, and small venous pressures decreased significantly. The onset of cardiovascular decompensation was associated with hyperpolarization (-53 +/- 3 mV), vasodilation, and return of venous pressure to control levels. Although direct electrical stimulation of the VSM and norepinephrine suffusion still produced constriction late in the hypotensive period, venoconstrictor responses to perivascular nerve stimulation failed progressively. This study indicates that failure of adrenergic neuromuscular transmission contributes significantly to the loss of compensatory VSM tone during hemorrhage.

Animals↗
PubMed1979

Estrogen receptors and effects of estrogen on membrane electrical properties of coronary vascular smooth muscle.

The effect of estrogen stimulation in vitro on the electrical properties of vascular smooth muscle (VSM), and the concentration of estrogen receptors in VSM were measured in isolated coronary arteries. Microelectrode measurements of the dog coronary artery membrane potential (Em) showed quiescent values of -51 millivolts (mV) and an input resistance (rin) of 10 megohms. Addition by diethylstilbestrol (DES) at 10(-6) M hyperpolarized the membrane to -64 mV and reduced input resistance (rin) to 5 megohms within 15 minutes. Extrapolation of the Em vs. log [K]o curve to zero potential gave similar values of [K]i of around 170 mM in both normal and DES treated muscles suggesting that the DES induced hyperpolarization is not due to increased Na-K pump activity. The 0.5% ethanol vehicle alone had no effect on the membrane potentials. Tetraethylammonium ion (TEA) induced action potentials in the previously quiescent tissue. When DES was applied in the presence of TEA, the membrane potential increased and the action potentials were abolished. Scatchard analysis of the estrogen receptor binding demonstrated both a high and a low affinity receptor for estrogen in the VSM. These data indicate that DES hyperpolarizes the VSM cells by a mechanism other than an increased Na-K pump activity. The mechanism of this increased Em may be due to factors which increase K+ conductance either mediated directly through estrogen interaction with its cytosolic receptors or through some unidentified second mechanism.

Action Potentials↗
PubMed1979

Action potential generation in reaggregates of rat aortic smooth muscle cells in primary culture.

Cultured vascular smooth muscle cells were prepared from adult rat aortas. The cells were dispersed by collagenase/elastase, and allowed to reaggregate into small spheres (50-200 micron in diameter) by plating on to cellophane. These primary cultures were incubated for 5-14 days, and then impaled with microelectrodes. The mean resting potential was -55 mV, and the mean input resistance was 9.0 M omega. The cells were quiescent, electrically and mechanically, and electrical stimulation usually did not elicit responses. However, addition of Ba++ (1 mM) or tetraehtylammonium ion (TEA; 5-15 mM) induced excitability (with accompanying contractions), either as spontaneous action potentials or by allowing responses to electrical stimulation. The cells became partially depolarized (e.g., to -36 mV) by these agents, and the input resistance increased. The frequency of spontaneous firing of the Ba++-induced spikes was affected by polarizing current pulses, as expected for pacemaker behavior. Elevation of Ca++ in the bathing solution increased the amplitude (overshoot) of the action potentials, and the spikes were blocked by verapamil (10(-5) M). Electron microscopy showed that the reaggregates consisted of a tight packing of elongated small-diameter cells, some of which exhibited thick and thin myofilaments and 'dense bodies'; many cells possessed surface caveolae. The results of this study demonstrate that reaggregates of arterial smooth muscle in primary culture can maintain functional and morphological characteristics of intact arterial smooth muscle, and therefore provide a useful preparation for the study of vascular smooth muscle function and control.

Action Potentials↗
PubMedDate not supplied

Bepridil blockade of Ca2+-dependent action potentials in vascular smooth muscle of dog coronary artery.

The effect of the new vasodilatory and antianginal compound, bepridil (CERM-1978), was examined on the electrical activity of the vascular smooth muscle of isolated dog coronary arteries. Tetraethylammonium (10 mM) was used to induce excitability in the muscle in the form of Ca2+-dependent overshooting action potentials, whose inward current is carried almost exclusively by Ca2+ ion through voltage-dependent slow channels. Bepridil (5 X 10(-7)--1 X 10(--5) M) produced a dose-dependent depression of the rate of rise and amplitude of these Ca2+ spikes. Complete blockade of the action potentials occurred at 1 X 10(-5) M bepridil. These effects of bepridil were antagonized by elevation of external Ca2+ concentration ([CA]o). The effects of bepridil were substantially reversed by washout after about 30 min. Bepridil (10(-5) M) also produced a small but significant (p less than 0.05) increase in resting membrane resistance (input resistance increased from a mean of 10.1 to 12.4 m omega), accompanied by a small but significant (p less than 0.05) depolarization of 6 mV (from a mean of --51 to --45 mV). These latter effects are consistent with a diminution of the resting K+ conductance (gK) by bepridil. It is concluded that the vasodilatory and antianginal properties of bepridil may be explained by the action of this drug in depressing and blocking the Ca2+ influx into the cells, presumably by acting directly on the voltage-dependent slow channels in the cell membrane, and thereby lowering [Ca]i and thus the degree of contraction. Bepridil has Ca2+-antagonistic (or Ca2+ entry blocking or slow channel blocking) properties much like verapamil, but it is somewhat less potent than verapamil in this action (i.e., complete blockade occurred at 10(-5) M bepridil vs. 2 X 10(-6) M verapamil).

Action Potentials↗