Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Electric Capacitance”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,639 records · Page 91Linked to original sources
PubMed1996

Some practical biological phantoms for calibrating multifrequency electrical impedance tomography.

Three groups of materials have been assessed with a Hewlett-Packard 4284A impedance analyser and Sheffield Mark 1 electrical impedance tomography (EIT) system for suitability for calibration of multifrequency EIT systems. They were required to be easy to use, stable over several hours, and have complex impedance similar to biological tissue. The groups were: (i) inorganic materials including barium titanate, polystyrene microspheres and fumed silica, all in aqueous suspension; these had phase angles below 1 degrees and so were unsuitable. (ii) Cucumber in KCl solution. Cucumber cortex had a phase angle of 40 degrees at a centre frequency of 50 kHz. Contrast between the cucumber and bathing solution could be selected by varying the KCl concentration. (iii) Polyurethane sponge immersed in packed red cells. The phase angle of packed cells was about 25 degrees at 1 MHz. Sponge resistivities and permittivities when immersed in packed cells were 5-20% higher than the bathing solution itself, for densities of 2-6.2% w/v. Both the biological materials appear suitable for the intended purpose; system (iii) is inherently more stable, and has capacitance in both bathing medium and test object. If an initial accuracy of greater than about +/- 15% is required, direct measurement with an impedance analyser is recommended prior to imaging studies.

Electric Impedance↗
PubMed1994

Dielectric spectroscopy of human erythrocytes: investigations under the influence of nystatin.

When placed in rotating electric fields red blood cells show a typical electrorotation spectrum with antifield rotation in the lower and cofield rotation in the higher frequency range. Assuming a spherical cell geometry, however, dielectrical parameters were obtained that differ from those measured by independent methods. Dielectrophoresis and, in particular, electrorotation yielded lower membrane capacitance values than expected. Introduction of an ellipsoidal model with an axis ratio of 1:2 allowed a description that proved to be consistent with dielectrophoresis and electrorotation data. For control cells an internal conductivity of 0.535 S/m, a specific membrane capacitance of 0.82 x 10(-2) F/m2, and a specific conductance of 480 S/m2 were obtained. The first characteristic frequency (frequency of fastest antifield rotation) and the related rotation speed can be measured quite quickly by means of a compensation method. Thus it was possible to follow changes of dielectric properties on individual cells after nystatin application. Ionophore-membrane interaction caused cell shrinkage in parallel to a decrease of the first characteristic frequency and rotation speed. Analysis of data revealed a decrease of the internal conductivity that is not only caused by ion loss but also, to a large extent, by a strong increase of hindrance because of shrinkage. Ionophore-induced membrane permeabilities can be calculated from volume decrease as well as from electrorotational data. In no case can these permeabilities count for the high membrane-AC conductivity that is attributed to the band-3 anion exchanging protein. The membrane-AC conductance was found not to be decreased for cells in Donnan equilibrium, which had leaked out almost completely.

Biophysical Phenomena↗
PubMed2001

Post-cultured development of basic electrophysiological properties of spinal neurons obtained from rat embryo.

Basic electrical profiles of cultured neurons are modified by multiple factors, such as cell growth, differentiation and cell damage from the isolation procedure. In the present study, we assessed development of electrophysiological properties of rat spinal neurons over the late embryonic and early postnatal period in a neuron-enriched culture. After recovery from acute damage within 2 days after plating, the input conductance and amplitudes of voltage-gated Na(+) and K(+) currents increased parallel to the increase in the cell capacitance. Whether this depended on the period or the growth of the cell area was estimated by normalizing the parameters with the cell capacitance. The input conductance per unit area, the membrane time constant and the Na(+) current density remained constant for two weeks. However, densities of two types of outward K(+) currents, an A-current and a delayed rectifier, required 3--5 days to reach the maximum, although neither thresholds for activation nor sensitivities to blockers (TEA and 4AP) altered. The hyperpolarizing shift of the resting membrane potential became stabilized within 6--8 days, suggesting that the development of the K(+) currents underlay the shift. These results show that passive electrical properties and voltage-gated currents of rat spinal neurons in the neuron-rich culture differ in temporal patterns of development but stabilize at the latest within a week, corresponding to the day of birth.

Animals↗
PubMed1984

Relations among passive electrical properties of lumbar alpha-motoneurones of the cat.

The relations among passive membrane properties have been examined in cat motoneurones utilizing exclusively electrophysiological techniques. A significant relation was found to exist between the input resistance and the membrane time constant. The estimated electrotonic length showed no evident tendency to vary with input resistance but did show a tendency to decrease with increasing time constant. Detailed analysis of this trend suggests, however, that a variation in dendritic geometry is likely to exist among cat motoneurones, such that the dendritic trees of motoneurones projecting to fast-twitch muscle units are relatively more expansive than those of motoneurones projecting to slow-twitch units. Utilizing an expression derived from the Rall neurone model, the total capacitance of the equivalent cylinder corresponding to a motoneurone has been estimated. With the assumption of a constant and uniform specific capacitance of 1 mu F/cm2, the resulting values have been used as estimates of cell surface area. These estimates agree well with morphologically obtained measurements from cat motoneurones reported by others. Both membrane time constant (and thus likely specific membrane resistivity) and electrotonic length showed little tendency to vary with surface area. However, after-hyperpolarization (a.h.p.) duration showed some tendency to vary such that cells with brief a.h.p. duration were, on average, larger than those with longer a.h.p. durations. Apart from motoneurones with the lowest values, axonal conduction velocity was only weakly related to variations in estimated surface area. Input resistance and membrane time constant were found to vary systematically with the a.h.p. duration. Analysis suggested that the major part of the increase in input resistance with a.h.p. duration was related to an increase in membrane resistivity and a variation in dendritic geometry rather than to differences in surface area among the motoneurones. The possible effects of imperfect electrode seals have been considered. According to an analysis of a passive membrane model, soma leaks caused by impalement injury will result in underestimates of input resistance and time constant and over-estimates of electrotonic length and total capacitance. Assuming a non-injured resting potential of -80 mV, a comparison of membrane potentials predicted by various relative leaks (leak conductance/input conductance) with those actually observed suggests that the magnitude of these errors in the present material will not unduly affect the presented results.+4

Animals↗
PubMed1984

Electrical properties of oligodendrocytes in culture.

The electrical properties of immunocytologically identified oligondendrocytes from embryonic mouse spinal cord maintained in culture for 3 to 6 weeks studied by passing current and recording potential changes with two separate intracellular electrodes. The average input resistance was 3.3 M omega and ranged from 0.7 to 16 M omega (n = 35). The input resistance increased by 19% with depolarization and decreased by 9% with hyperpolarization of 25 mV. The membrane time constant determined from the slope of the late exponential tail was 3.45 +/- 2.5 ms SD (n = 15). The specific membrane resistance of three cells was determined by a simplified square pulse analysis combined with measurement of membrane area. Membrane area was estimated from photomicrographs of cells injected with Lucifer Yellow CH and stained with the cell surface-reactive antibody 04 and from electron micrographs. An average specific membrane resistance of 1.3 X 10(3) omega cm2 and specific capacitance of 1.7 mu F/cm2 were calculated. Increasing [K+]o depolarized the cells and decreased the input resistance and the time constant.

Animals↗
PubMed1993

Does isoflurane alter mesenteric venous capacitance in the intact rabbit?

Volatile anesthetics act at a number of sites to alter cardiovascular function and the response of the cardiovascular system to barostatic reflexes. We examined the effects of isoflurane on reflex regulation of mesenteric venous capacitance vessels. To determine whether isoflurane alters mesenteric venous capacitance, continuous direct observations of mesenteric vein diameter, intravenous pressure, and mesenteric sympathetic efferent nerve activity (SENA) were made in 31 chloralose-anesthetized New Zealand white rabbits. Simultaneous measurements were obtained for aortic pressure and heart rate. The responses to changes in baroreceptor activation by means of either bilateral carotid occlusion (BCO) or aortic nerve stimulation (ANS) were studied in one group of 18 rabbits, while the response to direct electric activation by means of celiac ganglion stimulation (CGS) was studied in another group of 13 rabbits. In both groups, isoflurane vapor was administered at levels of 0.75% or 1.5%, and superfused isoflurane was administered directly to the vessel in doses of either 3% or 5% equilibrated with physiologic salt solution. Anesthetic levels were verified by mass spectrometry for expired gas and by gas chromatography for blood and superfusate levels. Inhaled isoflurane reduced hemodynamic variables and SENA in a dose-dependent fashion, but these same variables were unaffected by superfused isoflurane. One and one-half percent inhaled isoflurane abolished all reflex responses to baroreceptor stimulation in mesenteric capacitance veins and in SENA, but superfused isoflurane produced no corresponding attenuation of reflex responses to baroreceptor stimulation. Neither inhaled nor superfused isoflurane suppressed the reflex venoconstriction in response to CGS. Both inhaled and superfused isoflurane dilated the baseline vein diameter before stimulation. These results indicate that isoflurane dose increase the diameter of mesenteric venous capacitance vessels and inhibits reflex-induced constriction of mesenteric veins, whereas mesenteric sympathetic efferent nerve activity decreases and the reflex responses to activation of the carotid sinus and aortic baroreceptors are attenuated by inhaled isoflurane. The mechanism of this action appears to be primarily through the inhibition of central or peripheral sympathetic ganglionic transmission of barostatic control.

Animals↗
PubMed1994

Physiology, morphology and detailed passive models of guinea-pig cerebellar Purkinje cells.

1. Purkinje cells (PCs) from guinea-pig cerebellar slices were physiologically characterized using intracellular techniques. Extracellular caesium ions were used to linearize the membrane properties of PCs near the resting potential. Under these conditions the average input resistance, RN, was 29 M omega, the average system time constant, tau 0, was 82 ms and the average cable length, LN, was 0.59. 2. Three PCs were fully reconstructed following physiological measurements and staining with horseradish peroxidase. Assuming that each spine has an area of 1 micron 2 and that the spine density over the spiny dendrites is ten spines per micrometre length, the total membrane area of each PC is approximately 150,000 microns 2, of which approximately 100,000 microns 2 is in the spines. 3. Detailed passive cable and compartmental models were built for each of the three reconstructed PCs. Computational methods were devised to incorporate globally the huge number of spines into these models. In all three cells the models predict that the specific membrane resistivity, Rm, of the soma is much lower than the dendritic Rm (approximately 500 and approximately 100,000 omega cm2 respectively). The specific membrane capacitance, Cm, is estimated to be 1.5-2 muF cm-2 and the specific cytoplasm resistivity, Ri, is 250 omega cm. 4. The average cable length of the dendrites according to the model is 0.13 lambda, suggesting that under caesium conditions PCs are electrically very compact. Brief somatic spikes, however, are expected to attenuate 30-fold when spreading passively into the dendritic terminals. A simulated 200 Hz train of fast, 90 mV somatic spikes produced a smooth 12 mV steady depolarization at the dendritic terminals. 5. A transient synaptic conductance increase, with a 1 nS peak at 0.5 ms and a driving force of 60 mV, is expected to produce approximately 20 mV peak depolarization at the spine head membrane. This EPSP then attenuates between 200- and 900-fold into the soma. Approximately 800 randomly distributed and synchronously activated spiny inputs are required to fire the soma. 6. The passive model of the PC predicts a poor resolution of the spatio-temporal pattern of the parallel fibre input. An equally sized, randomly distributed group of approximately 1% of the parallel fibres, activated within a time window of a few milliseconds, would result in approximately the same composite EPSP at the soma.

Animals↗
PubMed1997

Scanning Single-Electron Transistor Microscopy: Imaging Individual Charges

A single-electron transistor scanning electrometer (SETSE)-a scanned probe microscope capable of mapping static electric fields and charges with 100-nanometer spatial resolution and a charge sensitivity of a small fraction of an electron-has been developed. The active sensing element of the SETSE, a single-electron transistor fabricated at the end of a sharp glass tip, is scanned in close proximity across the sample surface. Images of the surface electric fields of a GaAs/AlxGa1-xAs heterostructure sample show individual photo-ionized charge sites and fluctuations in the dopant and surface-charge distribution on a length scale of 100 nanometers. The SETSE has been used to image and measure depleted regions, local capacitance, band bending, and contact potentials at submicrometer length scales on the surface of this semiconductor sample.

Journal Article↗
PubMed1990

Inward rectification in the inner segment of single retinal cone photoreceptors.

1. Single cone photoreceptors were dissociated from the retina of a lizard with the aid of papain. The majority of the cells lost their outer segments but had well-preserved, large synaptic pedicles. Electrical properties of the cells were studied with tight-seal electrodes in the whole cell configuration. On the average, cone inner segments had a resting potential of -55 mV, and at this potential their input resistance was 2.6 G omega and their capacitance was 8 pF. 2. Under current clamp the cones exhibited a pronounced anomalous voltage rectification in response to hyperpolarizing currents. The voltage rectification was eliminated by external Cs+. 3. The Cs(+)-sensitive current underlying voltage rectification was isolated by blocking other currents present in the cone. Co2+ blocked a voltage-dependent Ca2+ current and a Ca2(+)-dependent Cl- current, and tetraethylammonium (TEA)+ blocked a delayed-rectifier K+ current. 4. The Cs(+)-sensitive current was activated by hyperpolarization to potentials more negative than -50 mV, and its current-voltage (I-V) relationship exhibited inward rectification. 5. The inward-rectifying current was selective for K+, but not exclusively. Increasing external K+ concentration 10-fold shifted the reversal potential by 13 mV. If Na ions also permeate through the inward-rectifying channels, the ratio of permeabilities (PK+/PNa+) in normal solution is approximately 3.9. 6. The kinetics of the inward-rectifying current were described by the sum of two exponentials, the amplitudes and time constants of which were voltage dependent. 7. The voltage dependence of the inward-rectifying current was described by Boltzmann's function, with half-maximum activation at -79 mV and a steepness parameter of 7.5 mV. 8. The voltage dependence and kinetics of the inward-rectifying current suggest that it is inactive in a cone photoreceptor in the dark. However, it becomes activated in the course of large hyperpolarizations generated by bright-light illumination. This activity will modify the waveform of the photovoltage--the current will generate a depolarizing component that opposes the light-generated hyperpolarization.

Animals↗
PubMed2001

Hyperinsulinism of infancy: the regulated release of insulin by KATP channel-independent pathways.

Hyperinsulinism of infancy (HI) is a congenital defect in the regulated release of insulin from pancreatic beta-cells. Here we describe stimulus-secretion coupling mechanisms in beta-cells and intact islets of Langerhans isolated from three patients with a novel SUR1 gene defect. 2154+3 A to G SUR1 (GenBank accession number L78207) is the first report of familial HI among nonconsanguineous Caucasians identified in the U.K. Using patch-clamp methodologies, we have shown that this mutation is associated with both a decrease in the number of operational ATP-sensitive K+ channels (KATP channels) in beta-cells and impaired ADP-dependent regulation. There were no apparent defects in the regulation of Ca2+- and voltage-gated K+ channels or delayed rectifier K+ channels. Intact HI beta-cells were spontaneously electrically active and generating Ca2+ action currents that were largely insensitive to diazoxide and somatostatin. As a consequence, when intact HI islets were challenged with glucose and tolbutamide, there was no rise in intracellular free calcium ion concentration ([Ca2+]i) over basal values. Capacitance measurements used to monitor exocytosis in control and HI beta-cells revealed that there were no defects in Ca2+-dependent exocytotic events. Finally, insulin release studies documented that whereas tolbutamide failed to cause insulin secretion as a consequence of impaired [Ca2+]i signaling, glucose readily promoted insulin release. Glucose was also found to augment the actions of protein kinase C- and protein kinase A-dependent agonists in the absence of extracellular Ca2+. These findings document the relationship between SUR1 gene defects and insulin secretion in vivo and in vitro and describe for the first time KATP channel-independent pathways of regulated insulin secretion in diseased human beta-cells.

ATP-Binding Cassette Transporters↗
PubMed1985

Passive water flows driven across the isolated rabbit ileum by osmotic, hydrostatic and electrical gradients.

Water flows generated by osmotic and hydrostatic pressure and electrical currents were measured in sheets of isolated rabbit ileum at 20 degrees C. Flows across the mucosal and serosal surfaces were monitored continuously by simultaneous measurement of tissue volume change (with an optical lever) and net water flows across one surface of the tissue (with a capacitance transducer). Osmotic gradients were imposed across the mucosal and serosal surfaces of the tissue separately, using probe molecules of various sizes from ethanediol (68 Da) to dextrans (161 000 Da). Flows across each surface were elicited with very short delay. The magnitudes of the flows were proportional to the osmotic gradient and related to the size of the probe molecule. Osmotic flow across the mucosal surface was associated with streaming potentials which were due to electro-osmotic water flow. The mucosal surface is a heteroporous barrier with narrow (0.7 nm radius, Lp (hydraulic conductivity) = (7.6 +/- 1.6) X 10(-9) cm s-1 cmH2O-1) cation-selective channels in parallel with wide neutral pores (ca. 6.5 nm radius, Lp = (2.3 +/- 0.2) X 10(-7) cm s-1 cmH2O-1) which admit large pressure-driven backflows from the submucosa to the lumen. There is additional evidence for a further set of narrow electroneutral pores less than 0.4 nm radius with Lp less than 7 X 10(-9) cm s-1 cmH2O-1. The serosal surface has neutral pores of uniform radius (ca. 6.5 nm), Lp = (7.6 +/- 1.6) X 10(-8) cm s-1 cmH2O-1. Hypertonic serosal solutions (100 mM-sucrose) cause osmotic transfer of fluid from isotonic mucosal solutions into the submucosa, expand it, and elevate the tissue pressure to 19.6 +/- 3.2 cmH2O (n = 4). Conversely, hypertonic mucosal solutions (100 mM-sucrose) draw fluid out of the submucosa in the presence of isotonic serosal solutions, collapse the submucosa, and lower the tissue pressure to -87.7 +/- 4.6 cmH2O (n = 5). Water flows coupled to cation movement could be generated across the mucosal surface in both directions by brief direct current pulses. The short latency of onset and cessation of flow (less than 2 s), absence of polarization potentials, and high electro-osmotic coefficients (range 50-520 mol water F-1), together with the presence of streaming potentials during osmotically generated water flows indicate electro-osmotic water flow through hydrated channels in the tight junctions and/or lateral intercellular spaces.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1996

Impedance spectra of tumour tissue in comparison with normal tissue; a possible clinical application for electrical impedance tomography.

Electrical characteristics of living tissues have been investigated for a long time in the search for further methods to complement the traditional investigations of pathology and physiology. Tumour tissue has been shown to exhibit a larger permittivity and conductivity than normal tissues. This might be associated with the fact that tumour cells have a higher water content and sodium concentration than normal cells, as well as different electrochemical properties of their cell membranes. To our knowledge only a few contributions on this subject have been published. This study describes an additional application on measurements of the complex impedance of tumour and normal tissues, in order to compare the impedance features of the two tissue types. The tissue sample is placed in a measuring cell in which the temperature is controlled. The measuring cell is connected to an impedance meter able to measure the complex impedance in terms of real and imaginary part curves for frequencies from 1.5 kHz to 700 kHz. The four-electrode principle is used with the current injected by the outer electrodes and the voltage difference measured between the inner electrodes. The current can be altered up to 1 mA. The instrument can be calibrated with known resistance and capacitance networks connected to the input of the instrument in order to minimize the measurement errors. The calibration routine uses a polynomial adaptation and can be applied interactively. Measurements performed by the instrument show promising results. Preliminary results show that this method can be extended to a new application for detection of tumour tissue by electrical impedance tomography (EIT).

Animals↗
PubMed1999

L-type calcium current and contractility in ventricular myocytes from mice overexpressing the cardiac beta 2-adrenoceptor.

OBJECTIVES: The reported increase in basal activity of hearts from transgenic mice (TG4) overexpressing the human beta 2-adrenoceptor (beta 2-AR) was explained by spontaneously active beta 2-ARs that stimulate the beta-adrenergic cascade in the absence of an agonist. In order to examine altered myocardial function on a cellular level, we have investigated L-type calcium current (ICa,L) and cell shortening in ventricular myocytes from TG4 hearts. Myocytes from littermates (LM) and wild type animals (WT) served as controls. METHODS: Cardiac beta-AR density was measured by [125I]-iodocyanopindolol binding to ventricular membranes. ICa,L was assessed by standard whole-cell voltage clamp technique. Contractility was measured as cell shortening in ventricular myocytes and as force of contraction in electrically stimulated left atria. RESULTS: Overexpression of beta 2-ARs was confirmed by an almost 400-fold increase in beta-AR density. The beta 1:beta 2-AR ratio in WT mice was 71:29. Myocytes from TG4 and LM mice were similar in size as judged by membrane capacitance and two dimensional cell area. ICa,L amplitude was significantly lower in TG4 than in LM myocytes (with 2 mM [Ca2+]o -4.82 +/- 0.48 vs. -6.56 +/- 0.38 pA/pF, respectively). In TG4 myocytes, the ICa,L response to isoproterenol (1 microM) was almost abolished. Cell shortening was not different in physiological [Ca2+]o, but smaller in maximum [Ca2+]o when comparing TG4 to control myocytes. Basal force of contraction in left atria did not differ between TG4 and LM at any age investigated. In TG4 left atria the inotropic response to isoproterenol was also absent, whereas responses to high [Ca2+]o or dibutyryl-cAMP (1 mM) were present but reduced. The rate of spontaneous beating of right atria was elevated in TG4 mice. CONCLUSIONS: Since only spontaneous beating rate but neither basal ICa,L amplitude nor basal contractile activity were elevated, our data fail to reveal evidence for spontaneously active, stimulating beta 2-ARs in left atrium and ventricle. A contractile deficit unrelated to the beta-adrenoceptor pathway is evident in TG4 myocytes and left atria.

Adrenergic beta-Agonists↗
PubMed1990

Two-frequency impedance plethysmograph: real and imaginary parts.

A four-channel impedance plethysmograph has been designed. Impedance signals are obtained at two frequencies by measuring both real and imaginary parts. Particular attention has been paid to the sine wave generation circuits that provide system versatility. The required phase-sensitive demodulation is achieved by means of analogue multiplexers. Results show that there are significant variations in the thoracic equivalent capacitance related to respiration and that there is an increased sensitivity to cardiac activity at low frequencies.

Analog-Digital Conversion↗
PubMed1979

Bacteriorhodopsin in model membranes. A new component of the displacement photocurrent in the microsecond time scale.

A quasi-short-circuit (tunable voltage clamp) measurement method with microsecond time resolution was applied to a bacteriorhodopsin model membrane formed by a novel interfacial technique. A new component (B1) of the displacement photocurrent was recorded: it has no detectable latency at an instrumental time constant of 1.5 museconds, and persists at 5 degrees C. In addition, a slower component (B2) of opposite polarity inhibited by low temperature (5 degrees C) and low pH (pH = 3.0) was recorded. The technique is very sensitive for the study of fast capacitative photoresponses in model membranes, and allows the detection of charge displacements in bacteriorhodopsin associated with distinct stages of the photochemical transformation.

Bacteriorhodopsins↗
PubMed1999

Factors influencing capacitance-based monitoring of microbial growth.

Microbiological impedance devices are routinely used by food and manufacturing industries, and public health agencies to measure microbiological growth. Factors contributing to increases and decreases in capacitance at the culture medium-electrode interface are poorly understood. To objectively evaluate the effects of temperature, cell density and medium conductivity on capacitance, admittance values from an impedance device were standardized; capacitance was converted to susceptance to allow unit comparisons with conductance. Although increases in temperature increased susceptance, a linear relationship could not be established between the change of susceptance with temperature and conductance of the medium. Cell density by itself had no measureable effect on susceptance or conductance, indicating that cells did not impede the movement of ions in the medium or around the electrode. In a low conductivity medium, increases in conductance by the addition of ions resulted in a concomitant increase of susceptance values. However, in a high conductivity medium, increases in conductance resulted in little or no increase of susceptance values because ions saturated the electrode surface. Susceptance increased when Escherichia coli, Pseudomonas aeruginosa, Alcaligenes faecalis and Staphylococcus aureus were grown in high conductivity media because protons produced by metabolically active bacteria balance more charge on the electrode than other ions. Increases in susceptance due to bacterial growth and metabolism in low conductivity media were attributed to both increases in protons and ionic metabolites. These results indicate that capacitance may provide a better measure of microbial growth and metabolism than conductance.

Bacteria↗
PubMed1999

Capacitative calcium entry in smooth muscle cells from preglomerular vessels.

Calcium entry via voltage-gated L-type channels is responsible for at least half of the increase in cytosolic calcium ([Ca(2+)](i)) in afferent arterioles following agonist stimulation. We sought the presence of capacitative calcium entry in fresh vascular smooth muscle cells (VSMC) derived from rat preglomerular vessels. [Ca(2+)](i) was measured using fura-2 ratiometric fluorescence. Vasopressin V1 receptor agonist (V1R) (10(-7) M) increased [Ca(2+)](i) by approximately 100 nM. A calcium channel blocker (CCB), nifedipine or verapamil (10(-7) M), inhibited the response by approximately 50%. V1R in the presence of CCB increased [Ca(2+)](i) from 106 to 176 nM, confirming that calcium mobilization and/or entry may occur independent of voltage-gated channels. In nominally Ca(2+)-free buffer, V1R increased [Ca(2+)](i) from 94 to 129 nM, denoting mobilization; addition of CaCl(2) (1 mM) further elevated [Ca(2+)](i) to 176 nM, indicating a secondary phase of Ca(2+) entry. Similar responses were obtained when CCB was present in calcium-free buffer or when EGTA was present. In nominally Ca(2+)-free medium, the sarcoplasmic reticulum Ca(2+)-ATPase inhibitors (SRCAI), thapsigargin and cyclopiazonic acid (CPA), increased [Ca(2+)](i) from 97 to 128 and 143 nM, respectively, and to 214 and 220 nM, respectively, when 1 mM extracellular Ca(2+) was added. In the presence of verapamil, the results with CPA acid were nearly identical. In Ca(2+)-free buffer, the stimulatory effect of V1R or SRCAI on the Ca(2+)/fura signal was quenched by the addition of Mn(2+) (1 mM), demonstrating divalent cation entry. These studies provide evidence for capacitative (store- operated) calcium entry in VSMC freshly isolated from rat preglomerular arterioles.

Animals↗
PubMed1985

Pump currents generated by the purified Na+K+-ATPase from kidney on black lipid membranes.

The transport activity of purified Na+K+-ATPase was investigated by measuring the electrical pump current induced on black lipid membranes. Discs containing purified Na+K+-ATPase from pig kidney were attached to planar lipid bilayers in a sandwich-like structure. After the addition of only microM concentrations of an inactive photolabile ATP derivative [P3-1-(2-nitro)phenylethyladenosine 5'-triphosphate, caged ATP] ATP was released after illumination with u.v.-light, which led to a transient current in the system. The transient photoresponse indicates that the discs and the underlying membrane are capacitatively coupled. Stationary pump currents were obtained after the addition of the H+, Na+ exchanging agent monensin together with valinomycin to the membrane system, which increased the permeability of the black lipid membrane for the pumped ions. In the absence of ADP and Pi the half saturation for the maximal photoeffect was obtained at 6.5 microM released ATP. The addition of ADP decreased the pump activity. Pump activity was obtained only in the presence of Mg2+ together with Na+ and Na+ and K+. No pump current was obtained in the presence of Mg2+ together with K+. The electrical response was blocked completely by the Na+K+-ATPase-specific inhibitors vanadate and ouabain. No pump currents were observed with a chemically modified protein, which was labelled on the ATP binding site with fluoresceine isothiocyanate. The method described offers the possibility of investigating by direct electrical measurements ion transport of Na+K+-ATPase with a large variety of different parameters.

Animals↗