Fernando Garcia Diaz

Boston University | BU · Department of Physiology and Biophysics

Neurons of the cochlear ganglion are endowed with a set of voltage-gated ion channels that enable them to encode and transmit sound information from the cochlear receptors to the brain. The temporal expression pattern of the K+ currents in chick cochlear ganglion neurons during embryonic development was analyzed using whole-cell voltage clamp techn...

Calcium entry through voltage-dependent channels play a critical role in neuronal development. Using patch-clamp techniques we have identified the components of the macroscopic Ca2+ current in acutely-isolated chick cochlear ganglion neurons and analysed their functional expression throughout embryonic development. With Ba2+ as a charge carrier, th...

The aim of our study was to define the effects of acidosis on the contractility of trabecular smooth muscle. Rabbit corpus cavernosal strips were mounted in organ chambers to measure isometric tension. Additionally, intracellular free Ca2+ concentration ([Ca2+]i) and tension were measured simultaneously utilising the intracellular fluorescent dye,...

Studies of glucose transporter activity and anti-glucose transporter (GLUT1) immunoblots were performed on different endothelial cell primary cultures (brain capillary, adrenal capillary and aortic) to determine their response to glucose deprivation. Cell cultures were exposed to glucose deprivation (0.5 mM) for 48 h periods and refed (11.0 mM) for...

To investigate the effects of severe hypoxia on trabecular smooth muscle contractility. Strips of rabbit corpus cavernosum were mounted in organ chambers to measure isometric tension. In some experiments intracellular free Ca2+ concentration and tension were measured by the intracellular fluorescent dye FURA-2 and isometric tension recording simult...

Whole-cell and single channel currents were studied in cells from frog (R. pipiens and R. catesbiana) skin epithelium, isolated by collagenase and trypsin treatment, and kept in primary cultures up to three days. Whole-cell currents did not exhibit any significant time-dependent kinetics under any ionic conditions used. With an external K gluconate...

Ionic gradients and fluxes are involved in a number of cellular functions including nerve and muscle excitation and transepithelial absorption and secretion. It is of critical importance in these cases to obtain accurate measurements of intracellular ionic activities both at rest and during cell activation. Since the miniaturization of ion-selectiv...

Knowledge of the voltage dependencies of apical and basolateral conductances is important in determining the factors that regulate transcellular transport. To gain this knowledge it is necessary to distinguish between cellular and paracellular currents and conductances. This is generally done by sequentially measuring transepithelial current/voltag...

Cell Na activity, acNa, was measured in the short-circuited frog skin by simultaneous cell punctures from the apical surface with open-tip and Na-selective microelectrodes. Skins were bathed on the serosal surface with NaCl Ringer and, to reduce paracellular conductance, with NaNO3, Ringer on the apical surface. Under control conditions acNa averag...

In studies of apical membrane current-voltage relationships, in order to avoid laborious intracellular microelectrode techniques, tight epithelia are commonly exposed to high serosal K concentrations. This approach depends on the assumptions that high serosal K reduces the basolateral membrane resistance and potential to insignificantly low levels,...

In voltage-clamped epithelia the cell membrane potential transient during a + 10-mV transepithelial pulse conforms to the expected behavior for a series combination of two linear resistance-capacitance (RC) circuits. The evolution of the cell potential is characterized by a single time constant with values of 30-130 ms in frog skin and Necturus gal...

Cell K activity,a k, was measured in the short-circuited frog skin by simultaneous cell punctures from the apical surface with open-tip and K-selective microelectrodes. Strict criteria for acceptance of impalements included constancy of the open-tip microelectrode resistance, agreement within 3% of the fractional apical voltage measured with open-t...

A device is described that permits continuous measurement of electrophysiological parameters in epithelial tissues in the open-circuit mode. Transepithelial potential (VT) and microelectrode (either conventional or ion-selective) potential (VM) are directly measured. Application of transepithelial current pulses allows continuous monitoring of tran...

It has been suggested that distribution of lateral interspace resistance in association with a highly conductive junction can significantly affect the measurement of outer membrane(o)/epithelial(t) voltage divider ratios (Fo = delta Vo/delta Vt), thereby leading to erroneous inferences regarding the outer membrane fractional resistance [fRo = Ro/Rc...

The absorption of water, electrolytes, and nutrient substances by the small intestine has been a central concern of gastrointestinal physiology for many years. Beginning as early as the mid-seventeenth century, there is a more or less continuous history of experimental investigation in this field (Parsons 1968). In this area, as in many other areas...

The epithelial and cellular effects of clamping the transepithelial potential (Vt, mucosa reference) have been investigated in the Necturus gallbladder. Following initial equilibration at short circuit, tissue conductance gt was 4.1 +/- 1.2 (SD) mS/cm2, the apical potential Va was -76 +/- 8 mV, and the apical fractional voltage on brief voltage per...

Open-tip and Cl–-selective microelectrodes were used to study the effect of external pH on apical membrane potential (V a) and intracellular chloride activity (a Cl i ) in epithelial cells ofNecturus gallbladder. Increasing the pH from 7.2 to 8.2 in the mucosal, the serosal, or in both bathing solutions simultaneously, hyperpolarizedV a (control...

As stated in the introduction, the purpose of this report has been to illustrate how the measurement of steady-state intracellular ionic activities, a technique that has proved to be of great importance in studying the energetics of transmembrane ionic transfer processes, can, with appropriate assumptions, be used to obtain information concerning t...

Intracellular Na+, K+, and Cl- activities (aiNa, aiK, aiCl) and transapical membrane potentials (V0) were measured with liquid ion-exchanger and open-tip microelectrodes in isolated short-circuited frog skins (R. pipiens) incubated at 23 degrees C in normal amphibian Ringer's solution. Under control conditions aiNa = 14 +/- 3 mM, aiK = 132 +/- 10 m...

This report reviews the use of ion-selective microelectrodes to measure intracellular ionic activities and ionic electrochemical potential differences across cell membranes. Particular emphasis is placed on the electrochemical characteristics of liquid ion-exchanger microelectrodes. Methods for assessing the effect of interfering ions on the electr...

Intracellular C1, K and Na activities (a Cl i ,a k i anda Na i ) and transmucosal membrane potential (E m) in epithelial cells ofNecturus gallbladder were measured at different external Na concentrations ([Na]o), with liquid ion-exchanger and conventional microelectrodes. Bladders were mounted in a divided chamber at 23C between identical...

Much evidence implicates the transapical Na+ electrochemical potential gradient as an energy source for the accumulative transfer of sugars, amino acids, and other solutes across the mucosal membrane of absorptive cells in the small intestine. Recently, this gradient has also been shown to be involved in electrically neutral Na+-coupled Cl-transpor...

The sodium-selective ligand 1,1,1-tris[1(1)-(2(1)-oxa-4(1)-oxo-5(1)-aza-5(1)-methyl)dodecanyl]propane dissolved in 3-nitro-o-xylene containing a small amount of the lipophilic anion tetrachlorophenyl borate was used as a liquid ion-exchanger in sodium-selective microelectrodes. The microelectrodes gave rapid, stable responses that were linear funct...

Na+, K+ and Cl- concentrations (cij) and activities (aij), and mucosal membrane potentials (Em) were measured in epithelial cells of isolated bullfrog (Rana catesbeiana) small intestine. Segments of intestine were stripped of their external muscle layers, and bathed (at 25 degrees C and pH 7.2) in oxygenated Ringer solutions containing 105 mM Na+ a...

Mucosal hypertonicity produces a drop of transepithelial potential difference in isolated rat jejunum with a half time of about 15 s. The same effect is obtained when the osmorality of both bathing solutions is raised simultaneously. Serosal hypertonicity produces an increase of transepithelial potential difference an order of magnitude lower than...