Biomedical subjects

D R Harder

Publications and source records attributed to D R Harder.

At least 73 records · Page 4Linked to original sources

PubMed1990

Enhanced vascular tone in the renal vasculature of spontaneously hypertensive rats.

The renal microvascular responses of Wistar-Kyoto and spontaneously hypertensive rats to changes in perfusion pressure were compared using a juxtamedullary nephron microvascular preparation perfused in vitro with a physiological salt solution containing 5% albumin. In the spontaneously hypertensive rats, the internal diameters of arcuate and interlobular arteries and the proximal and distal afferent arterioles averaged 307 +/- 26, 52 +/- 2, 24 +/- 0.9, and 22 +/- 1.2 microns, respectively, at 80 mm Hg. They were 18-35% smaller (p less than 0.05) than the corresponding vessels measured in Wistar-Kyoto rats. In low calcium media, the arcuate and interlobular arteries and the proximal and distal afferent arterioles of spontaneously hypertensive rats exhibited a greater dilation than the vessels of Wistar-Kyoto rats. These observations suggest that the diameters of the preglomerular vasculature of the spontaneously hypertensive rats are reduced because of an elevated vascular tone rather than structural changes narrowing the lumen of these vessels. These results suggest that enhanced vascular tone in the preglomerular vasculature of juxtamedullary nephrons may contribute to the elevated renal medullary vascular resistance and resetting of the pressure-natriuretic relation previously observed in spontaneously hypertensive rats.

Animals↗

PubMed1990

Role of the vascular endothelium in regulating the response of small arteries of the dog kidney to transmural pressure elevation and reduced PO2.

The goal of this study was to determine the role of the vascular endothelium in regulating the response of small renal arteries to increased transmural pressure and reduced PO2. Canine small renal arteries (496 +/- 32 microns) were isolated, cannulated with micropipettes, and mounted in an in vitro chamber that allowed transmural pressure, tissue bath PO2, and vessel lumen PO2 to be controlled. In intact arteries, elevation of transmural pressure caused contractile activation and a progressive depolarization (0.17 +/- 0.09 mV/mm Hg pressure increase) of the vascular smooth muscle cells. Endothelial damage by air perfusion caused a 13-14% reduction in resting diameter and an 18 +/- 3 mV depolarization of the vascular smooth muscle, but eliminated the progressive contraction and depolarization that occurred in intact vessels in response to transmural pressure elevation. Reduction of either tissue bath O2 concentration or vessel lumen PO2 significantly enhanced norepinephrine-induced contractions of the arteries. Endothelial damage prevented the enhancement of norepinephrine-induced contractions by reduced PO2 in the vessels. The results of this study suggest that the vascular endothelium regulates both myogenic contraction and the enhancement of norepinephrine-induced contractions by reduced PO2 in small arteries of the dog kidney.

Animals↗

PubMed1990

Endothelium-dependent modulation of endothelin-induced vasoconstriction and membrane depolarization in cat cerebral arteries.

Effects of endothelin, the newly discovered, endothelium-derived vasoactive peptide, were examined on isolated, pressurized cat arteries. Responses to increasing concentrations of porcine endothelin (10(-12) to 10(-8) M) were compared in endothelium intact and denuded vessel segments utilizing diameter and membrane potential measurements. Endothelin produced a concentration-dependent contraction in arteries with intact endothelium with a maximal response of 26 +/- 2.8% of the initial diameter in physiological saline solution at 100 mm Hg of intraluminal pressure. Removal of the endothelium sensitized the smooth muscle to endothelin and increased the maximal response to 48 +/- 2.3% of the initial diameter. After incubation of the intact vessels with 10(-5) M indomethacin, the endothelin response was potentiated to the maximal value of 66 +/- 1.26% of the initial diameter of the intact vessels, and to 61 +/- 2.6% of the denuded arteries. The arterial smooth muscle cells were depolarized by 6 +/- 0.45 mV after administration of endothelin to the vessels, and by 10 +/- 0.8 mV in the denuded vessels. This depolarization was not altered by indomethacin. Our data suggest that endothelin has a direct effect on the smooth muscle of cat cerebral vascular tissue, and the sensitivity of these vessels to endothelin may be modulated by the release of dilatory cyclooxygenase products derived from the intact endothelium.

Animals↗

PubMed1989

Pressure-dependent contraction of rat juxtamedullary afferent arterioles.

Pressure-diameter relations were studied in rat afferent arterioles using an isolated, juxtamedullary nephron preparation perfused with a saline solution containing 5% albumin. Angiotensin I (10 microM), angiotensin II (0.1 microM), and norepinephrine (10 microM) increased perfusion pressure, and norepinephrine, but not angiotensin I or II, contracted afferent arterioles, indicating that the vessels are reactive. The control diameter of the afferent arterioles that exhibited pressure-dependent contraction (n = 58) averaged 30.8 +/- 1.1 micron at perfusion pressure of 80 mm Hg. When pressure was increased from 80 to 120 and then to 180 mm Hg, the diameter of these arterioles decreased by 16.4 +/- 2.1%. Glomerular capillary pressure was well autoregulated and averaged 45.2 +/- 2.2, 50.2 +/- 2.4, and 53.0 +/- 3.0 mm Hg, respectively, at perfusion pressures of 80, 120, and 180 mm Hg. Administration of vasodilators or a Ca2+-free solution eliminated the contractile response to pressure elevations; rather, the diameter of these vessels increased significantly by 17.5 +/- 5.1% and 32.0 +/- 9.4%, respectively, when pressure was increased from 80 to 180 mm Hg. Blocking tubuloglomerular feedback mechanism, with furosemide or by removal of the renal papilla (which interrupts the delivery of fluid to the macula densa), eliminated the pressure-dependent contraction of the afferent arterioles. Instead the diameter of these vessels increased by 27.0 +/- 7.8% and 36.0 +/- 5.6%, respectively, when the pressure was increased from 80 to 120 and then to 180 mm Hg. These results demonstrate that juxtamedullary nephrons perfused in vitro autoregulate glomerular capillary pressure.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin I↗

PubMed1989

Pressure releases a transferable endothelial contractile factor in cat cerebral arteries.

When exposed to an increasing transmural pressure, middle cerebral arteries of the cat exhibit reduction of internal diameter which is mediated by vascular muscle cell depolarization. This laboratory has recently demonstrated that this "pressure-induced" activation is dependent upon the presence of an intact endothelium. The present studies were undertaken to determine if this phenomenon is due to inhibition of tonically released endothelium-derived relaxing factors (EDRF) or release of a contractile substance. When cerebral arterial segments were pressurized to between 40 and 160 mm Hg there was 13.2% reduction in internal diameter accompanied by significant muscle cell depolarization from -53 +/- 2.7 to -22 +/- 1.4 mV. There was a significant positive correlation between the delta Em and step increases in transmural pressure. These excitatory responses were lost and vessels dilated to pressure when the endothelium was removed. Upon exposing the denuded vessel to a pressurized intact donor, the denuded vessel recovered its ability to contract and depolarize suggesting that a contractile substance might be released from the vascular endothelium upon pressurization. The EDRF antagonist oxyhemoglobin did not alter the excitatory response to pressure in these isolated cerebral arteries further suggesting that the reduction in diameter and muscle cell depolarization results from the release of a contractile substance from the vascular endothelium and not inhibition of EDRF.

Animals↗

PubMed1989

Mechanism of action of EDRF on pressurized arteries: effect on K+ conductance.

Experiments were performed to study the cellular mechanism of endothelium-derived relaxing factor (EDRF) on vascular smooth muscle. Rat femoral arteries were cannulated and pressurized to 100 mm Hg. Vascular smooth muscle membrane potential (Em) and diameter responses to perfusion with 5 x 10(-6) M acetylcholine (ACh) were measured in vessels precontracted with 5 x 10(-6) M norepinephrine (NE). Hyperpolarization (-35 +/- 1.2 to -66 +/- 2.0 mV) and dilation were observed during ACh administration. Both responses were abolished on removal of the endothelium with collagenase. A bioassay was developed in which two vessel segments from the same artery were connected in series. The downstream vessel was deendothelialized while the endothelium of the upstream vessel remained intact. The protocol used was the same as in the first set of measurements. Hyperpolarization and dilation were observed in both vessels during ACh perfusion. However, when the direction of the perfusate flow in the bioassay system was reversed so that the deendothelialized vessel was upstream, only the "endothelium-intact" vessel demonstrated vascular smooth muscle hyperpolarization. To examine the ionic mechanism underlying the hyperpolarization presumably by released EDRF, the Em was measured as a function of increasing extracellular potassium ([K+]o). In the presence of ACh (but not NE) the maximum depolarization produced by a decade increase of [K+]o (10-100 mM) was 50 mV. In the deendothelialized vessel, this depolarization was decreased significantly to 39 mV. Addition to the superfusate of 10 mM tetraethylammonium, a K+ channel blocker, significantly reduced the hyperpolarization caused by ACh-induced EDRF release.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

PubMed1989

Mechanisms of pressure-induced myogenic activation of cerebral and renal arteries: role of the endothelium.

Control of blood flow is mediated by a variety of metabolic, neurogenic, myogenic and other mechanisms. The basic response of the myogenic mechanisms is a mechanical stimulus that activates vascular muscle via a sensor within the vessel wall. We studied the way in which increases in transmural pressure within the physiological range can activate cerebral and renal vascular muscle by depolarizing the muscle cell membrane. These membrane electrical effects depend on the external calcium concentration [( Ca2+]o) and on the presence of an intact vascular endothelium. Bioassay experiments have shown that an elevation in pressure releases a vasoactive product(s) from endothelial cells, which mediates the pressure-dependent membrane depolarization in these vessels. These responses may affect the autoregulation of blood flow in that vessels become smaller as pressures increase between 60 and 160 mmHg.

Animals↗

PubMed1988

Increased sensitivity of cat cerebral arteries to serotonin upon elevation of transmural pressure.

Segments of middle cerebral artery (MCA) were isolated from cat brains and cannulated allowing manipulation of transmural pressure (TP). These cannulated vessel segments were mounted in a specially fabricated myograph allowing measurement of internal diameter with the aid of a high resolution binocular microscope and a video imaging system. Internal diameter was then measured as a function of topically applied serotonin at 3 different levels of TP: 60, 100, and 140 mmHg. As TP was elevated from 60 to 140 mmHg the sensitivity to serotonin increased from an ED50 value of 1.3 x 10(-8) to 3.5 x 10(-10) M. We have yet to explore the mechanisms involved in the "pressure-mediated" increase in cerebrovascular sensitivity to serotonin; however, it may be related to the muscle membrane depolarization we have observed previously in response to elevations in TP. Such findings may account for the discrepancies in dose ranges for serotonin thought to be active in vivo vs. the higher concentrations needed to elicit responses in isolated vessels.

Animals↗

PubMed1988

Early effects of experimental diabetes on central catecholamine concentrations.

Six days after induction of diabetes norepinephrine (NE), 3,4-dihydroxyphenylalanine and dopamine concentrations were determined in spinal cord, cerebellum, pons/medulla oblongata and the remaining brain of diabetic rats. Only cerebellar NE levels were significantly increased by 21% compared to age-matched controls. This effect was not seen when diabetic rats received insulin replacement therapy. Fifty-two days after induction of diabetes cerebellar NE levels in diabetic rats were further increased (+37%), implying an enduring and progressive effect. The data show that even with a short period of uncontrolled diabetes central neurochemical alterations occur.

Animals↗

PubMed1987

Distribution of excitatory junction potential amplitude in cat cerebral arteries: examination of its quantal nature and modulation by opiates.

We used scorpion venom to release small amounts of an excitatory neurotransmitter from adventitial nerves in cat left anterior descending cerebral artery. We used glass microelectrodes to measure and record postsynaptic electrical events of minimal amplitude. These events were similar to postsynaptic spontaneous and electrically evoked excitatory junction potentials (ejp's) seen in skeletal muscle. We performed a frequency analysis of the ejp amplitudes to determine if they fit a unimodal or multimodal distribution. We also investigated the effects of phentolamine, norepinephrine, hydromorphone, and morphine on ejp amplitude and frequency in the artery. Statistical analysis of the ejp frequency and amplitude revealed a multipeaked distribution with decreasing peaks. These results were similar to the distribution reported for acetylcholine release in skeletal muscle. The ejps were inhibited by phentolamine, which suggested that these events were adrenergically mediated. Norepinephrine and the opiates, hydromorphone and morphine, reduced the frequency and amplitude of the ejp's. The vessels also constricted to increasing doses of norepinephrine both under control conditions and in the presence of opiate. These results suggest that norepinephrine blocks the ejp's by a feedback mechanism at the presynaptic membrane and that endorphins and/or enkephalins, also acting at this presynaptic site, may modulate neurotransmission in the cerebral circulation.

Animals↗

PubMed1987

The effect of alkaline pH and transmural pressure on arterial constriction and membrane potential of hypertensive cerebral arteries.

These studies were undertaken to examine the effect of alkalosis to modify "pressure-induced" activation of isolated cerebral arteries from spontaneously hypertensive rats (SHR) and their normotensive Wistar-Kyoto (WKY) controls. At pH 7.4 and PCO2 of 34 torr elevation of transmural pressure from 0-140 mm Hg resulted in myogenic activation preceded by membrane depolarization in both SHR and WKY. The degree of developed myogenic tone in SHR was elevated above WKY. Alkalosis (pH 7.4-7.7) depolarized and activated SHR cerebral arteries to a greater extent than WKY. Furthermore, both the electrical and mechanical responses to elevation in transmural pressure were exaggerated in SHR compared to WKY at pH 7.7 (PCO2 constant at 34 torr). Manipulation of PCO2 at constant pH of 7.4 had similar effects on "pressure-induced" myogenic tone in both SHR and WKY. Thus, cerebral arteries from both SHR and WKY depolarize and develop myogenic tone in response to increasing transmural pressure. This response is augmented in SHR, but to a much greater extent upon elevation of extracellular pH, while PCO2 is maintained within normal limits. The implications of these findings are discussed.

Animals↗

PubMed1987

Altered splenic catecholamine concentrations during experimental allergic encephalomyelitis.

Catecholamine concentrations of the spleen were studied with neurochemical techniques in rats injected with myelin basic protein to produce an experimental allergic encephalomyelitis (EAE). Thirteen to 14 days postinoculation the affected rats showed peak clinical signs of weakness, especially in the lower extremities. Resolution of the disease then progressed rapidly with full clinical recovery at day 21. Splenic concentrations of norepinephrine (NE), 3,4-dihydroxyphenylalanine (DOPA), epinephrine (EPI) and 3,4-dihyxroxyphenylethylamine (DA) were determined by HPLC with electrochemical detection. DOPA concentrations were significantly increased (+62%) while DA concentrations were decreased (-29%) in the EAE rats on day 14 postinoculation. NE and EPI concentrations tended to be elevated in the EAE group, but this was not statistically significant. No differences in splenic catecholamines were detected on day 7 and 52 postinoculation between EAE and control animals. These results indicate that changes in the metabolic pathways of splenic catecholamines occur at the peak of the clinical symptoms of EAE; the increase in DOPA and the decrease in DA concentrations suggest that the activity of DOPA-decarboxylase or its co-factor is altered.

Animals↗

PubMed1987

Effect of morphine on electrophysiological properties of circular and longitudinal muscles.

We have measured the effects of morphine on the intracellular electrophysiological properties of smooth muscle cells from the circular and longitudinal muscle layers of the canine jejunum. Morphine hyperpolarized the circular muscle membrane by approximately 12 mV and increased the electrical control activity (ECA) amplitude and dV/dt. Morphine had no significant effect on the electrical properties of the longitudinal muscle cells. The morphine-induced hyperpolarization of the circular muscle membrane was blocked by tetrodotoxin (TTX) and naloxone, but not by atropine and hexamethonium, propranolol, or phentolamine. Morphine significantly increased the slope of the resting membrane potential vs. the log of the potassium concentration in bathing medium from 38 to 50 mV/decade. The sodium permeability to potassium permeability ratio, calculated from the Goldman constant field equation, was reduced by morphine from 0.13 to 0.07 at mM of K+. The above results suggest that when measured by intracellular techniques, morphine hyperpolarizes the circular muscle membrane by release of a nonadrenergic, noncholinergic neurotransmitter. The mechanism of this hyperpolarization is consistent with an increase in potassium conductance.

Animals↗

PubMed1987

Vascular muscle cell depolarization and activation in renal arteries on elevation of transmural pressure.

The goal of this study was to define some of the cellular and ionic mechanisms of smooth muscle cell activation in dog renal arteries exposed to physiological levels of transmural pressure. Isolated interlobular arteries were cannulated and connected to a pressure reservoir to allow manipulation of transmural pressure in 20-mmHg increment steps from 20 to 120 mmHg. As transmural pressure was increased, vascular smooth muscle exhibited a linear depolarization from an average resting potential of -57 +/- 2 mV at 20 mmHg to -38 +/- 2.4 mV at 120 mmHg. Spontaneous action potentials could often be recorded at pressures greater than 80 mmHg. These appeared to occur primarily at bifurcation points of branching arteries. Vascular smooth muscle depolarization and action potentials occurring in response to increases in transmural pressure were associated with a maintenance of internal diameter of the vessel segments despite increases in transmural pressure in the range between 60 and 120 mmHg. The "pressure-induced" activation of vascular smooth muscle contraction and spontaneous action potentials of small renal arteries at higher transmural pressures were blocked on Ca2+ channel inhibition with verapamil (10(-6) M). These data document a membrane ionic mechanism (probably increased Ca2+ influx) for pressure-induced myogenic activation of isolated renal arteries. It is interesting that the contraction of these vessels occurs over the pressure range in which autoregulation of renal blood flow normally occurs. The physiological significance of these responses needs to be determined.

Animals↗

PubMed1987

Pressure-induced myogenic activation of cat cerebral arteries is dependent on intact endothelium.

These studies were designed to determine the role of cerebral vascular endothelium in the "myogenic" depolarization and contraction observed in isolated cat middle cerebral arteries exposed to high transmural pressures. With intact endothelial cells we observed, on elevation of transmural pressure in cannulated isolated arteries, significant membrane depolarization, action potential generation, and reduction in internal diameter. After perfusion of the same vessels with collagenase and elastase for short periods of time to disrupt the endothelial layer, all previous responses to elevation of transmural pressure were no longer seen. Even though enzyme perfusion had no effect on membrane potential at "control" levels of transmural pressure, it abolished the pressure-dependent depolarization, action potential generation, and constriction. Furthermore, the contractile response to agonist stimulation was maintained after endothelial disruption via enzymes, showing that this method of endothelial disruption did not appreciably damage muscle cells. The data document a dependence of an intact endothelium in mediating the activation of isolated cat cerebral arteries in response to a changing transmural pressure. Thus, it is possible that the endothelial cell may serve as a transducer in the autoregulatory response to pressure.

Action Potentials↗

PubMed1987

Electrical stimulation of the endothelial surface of pressurized cat middle cerebral artery results in TTX-sensitive vasoconstriction.

The purpose of this study was to examine the electrical and mechanical responses of cat middle cerebral arteries to electrical stimulation of the adventitial vs. intimal surface of the vessels and to determine the responses as a function of transmural pressure. Middle cerebral arteries were cannulated at both ends. Within each cannula was a stimulating electrode. Electrical stimulation (0.5-msec square current pulses at 0.5 Hz yielding 160 microA of current between electrodes) resulted in significant reduction in diameter that was greater at both 40 and 80 mm Hg vs. 100 or 140 mm Hg. Conversely, adventitial stimulation of perivascular nerves with transmural platinum stimulating electrodes resulted in significant vasodilation. The constrictor response to intimal stimulation, as well as the dilatory response to adventitial stimulation, was blocked by tetrodotoxin. The constrictor response to luminal stimulation was enhanced by scorpion toxin demonstrating a functional role for tissues containing fast Na+ channels. Perfusion with collagenase to disrupt the endothelium also abolished the constrictor response to luminal stimulation. The divergence of responses between adventitial and luminal surface stimulation may suggest that different cell layers within a blood vessel serve different functions, one to increase resistance and another to decrease resistance. For example, in cat middle cerebral arteries, the adventitial nerves (i.e., via reflexes) may increase flow, while blood-borne substances may mediate release of agents that reduce flow.

Animals↗

PubMed1987

Possible cellular mechanism for cerebral vasospasm after experimental subarachnoid hemorrhage in the dog.

This study was undertaken to examine some of the cellular ionic mechanisms responsible for the cerebral vasospasm that occurs as a consequence of subarachnoid hemorrhage (SAH). After cisternal injection of autologous blood we documented spasm of the basilar artery upon angiography from 4 to 7 d postictus in six dogs. When these basilar arteries were isolated we observed marked membrane depolarization and enhanced electrical spike activity compared with controls. The slope of the membrane potential vs log [K]0 curve was significantly reduced in arteries exposed to SAH. Further analysis supported the concept that such altered muscle cell properties resulted from reduction in resting K+ conductance (gk). Exposure of arteries in vitro to nicorandil (10(-9)-10(-7)M) (a drug which acts by increasing gk) hyperpolarized the muscle cells and increased internal diameter. Infusion of nicorandil (3-5 micrograms/kg per min) to intact, anesthetized animals reversed, by 50%, the reduction in basilar artery diameter after experimental SAH.

Action Potentials↗

PubMed1986

Effect of reduced oxygen availability upon myogenic depolarization and contraction of cat middle cerebral artery.

The goal of this study was to determine whether electrophysiological mechanisms contribute to the relaxation of cat middle cerebral artery in response to decreased ambient Po2 and whether decreased Po2 alters the myogenic depolarization and contraction of this vessel in response to elevations in transmural pressure. In one series of experiments, arterial segments (200-500 micron outer diameter) were isolated and mounted in an in vitro tension transducer to allow continuous measurement of active tension as bath Po2 was reduced. In these experiments, vessel relaxation occurred primarily between 150 mm Hg Po2 and 40 mm Hg Po2, suggesting that cerebral arteries are sensitive to alterations of Po2 in the physiological range. Relaxation did not result from the activation of dilator nerves in the vessel wall, since it was unaffected by tetrodotoxin. Arterial segments were also cannulated with micropipettes and subjected to elevations in transmural pressure during 300 mm Hg Po2 and 50 mm Hg Po2 superfusion. During 300 mm Hg Po2 superfusion, cannulated vessels exhibited myogenic depolarization and maintained their diameter as transmural pressure was increased; 50 mm Hg Po2 superfusion inhibited spontaneous spike activity, decreased the slope of the myogenic depolarization, and partially inhibited vessel contraction in response to elevated transmural pressure. These effects are independent of the parenchymal cell environment and appear to be mediated, at least in part, by electrophysiological mechanisms.

Animals↗