Proton Pump Activity of Mitochondria-rich Cells : The Interpretation of External Proton-concentration Gradients

We have hypothesized that a major role of the apical H+-pump in mitochondria-rich (MR) cells of amphibian skin is to energize active uptake of Cl− via an apical Cl−/HCO3−-exchanger. The activity of the H+ pump was studied by monitoring mucosal [H+]-profiles with a pH-sensitive microelectrode. With gluconate as mucosal anion, pH adjacent to the cornified cell layer was 0.98 ± 0.07 (mean ± SEM) pH-units below that of the lightly buffered bulk solution (pH = 7.40). The average distance at which the pH-gradient is dissipated was 382 ± 18 μm, corresponding to an estimated “unstirred layer” thickness of 329 ± 29 μm. Mucosal acidification was dependent on serosal pCO2, and abolished after depression of cellular energy metabolism, confirming that mucosal acidification results from active transport of H+. The [H+] was practically similar adjacent to all cells and independent of whether the microelectrode tip was positioned near an MR-cell or a principal cell. To evaluate [H+]-profiles created by a multitude of MR-cells, a mathematical model is proposed which assumes that the H+ distribution is governed by steady diffusion from a number of point sources defining a set of particular solutions to Laplace's equation. Model calculations predicted that with a physiological density of MR cells, the [H+] profile would be governed by so many sources that their individual contributions could not be experimentally resolved. The flux equation was integrated to provide a general mathematical expression for an external standing [H+]–gradient in the unstirred layer. This case was treated as free diffusion of protons and proton-loaded buffer molecules carrying away the protons extruded by the pump into the unstirred layer; the expression derived was used for estimating stationary proton-fluxes. The external [H+]-gradient depended on the mucosal anion such as to indicate that base (HCO3−) is excreted in exchange not only for Cl −, but also for Br− and I−, indicating that the active fluxes of these anions can be attributed to mitochondria-rich cells

Passive cation permeability of turtle colon: Evidence for a negative interaction between intracellular sodium and apical sodium permeability

The role of intracellular sodium in the regulation of apical sodium permeability was investigated in an electrically “tight” epithelium, the turtle colon. In the presence of low mucosal sodium (3 mM) and serosal ouabain, an inhibitor of the basolateral sodium pump, the apical membrane retained a substantial amiloride-sensitive, sodium conductance and the basolateral membrane exhibited a barium-sensitive potassium conductance in parallel with a significant sodium (and lithium) conductance. In the presence of a high mucosal sodium concentration (114 mM), however, inhibition of active sodium absorption by ouabain led to a disappearance of the amiloride-sensitive, transepithelial conductance that was due, at least in part, to a virtual abolition of the apical sodium permeability. Two lines of evidence indicate that this permeability decrease was dependent upon an increase in intracellular sodium content. First, raising the mucosal sodium concentration from 3–114 mM in the presence of ouabain reversibly inhibited the amiloride-sensitive conductance. The time course of the decline in conductance paralleled the apparent intracellular accumulation of sodium in exchange for potassium, which was monitored as a transient deflection in the amiloride-sensitive, short-circuit current. Second, the inhibitory effect of mucosal sodium-addition was markedly attenuated by serosal barium, which prevented the accumulation of sodium by blocking the electrically coupled, basolateral potassium exit. These results support the notion of a “negative feedback” effect of intracellular sodium on the apical sodium permeability.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47451/1/424_2004_Article_BF00583286.pd

Overview of the mars pathfinder mission : launch through landing, surface operations, data sheets, and science results

Mars Pathfinder successfully landed at Ares Vallis on July 4, 1997, deployed and navigated a small rover about l00 m clockwise around the lander, and collected data from three science instruments and ten technology experiments. The mission operated for three months and returned 2.3 Gbits of data, including over 16,500 lander and 550 rover images, 16 chemical analyses of rocks and soil, and 8.5 million individual temperature, pressure and wind measurements. Pathfinder is the best known location on Mars, having been clearly identified with respect to other features on the surface by correlating five prominent horizon features and two small craters in lander images with those in high-resolution orbiter images and in inertial space from two-way ranging and Doppler tracking. Tracking of the lander has fixed the spin pole of Mars, determined the precession rate since Viking 20 years ago, and indicates a polar moment of inertia, which constrains a central metallic core to be between 1300 and ~2000 km in radius. Dark rocks appear to be high in silica and geochemically similar to anorogenic andesites; lighter rocks are richer in sulfur and lower in silica, consistent with being coated with various amounts of dust. Rover and lander images show rocks with a variety of morphologies, fabrics and textures, suggesting a variety of rock types are present. Rounded pebbles and cobbles on the surface as well as rounded bumps and pits on some rocks indicate these rocks may be conglomerates (although other explanations are also possible), which almost definitely require liquid water to form and a warmer and wetter past. Airborne dust is composed of composite silicate particles with a small fraction of a highly magnetic mineral, interpreted to be most likely maghemite; explanations suggest iron was dissolved from crustal materials during an active hydrologic cycle with maghemite freeze dried onto silicate dust grains. Remote sensing data at a scale of a kilometer or greater and an Earth analog correctly predicted a rocky plain safe for landing and roving with a variety of rocks deposited by catastrophic floods, which are relatively dust free. The surface appears to have changed little since it formed billions of years ago, with the exception that eolian activity may have deflated the surface by ~3-7 cm, sculpted wind tails, collected sand into dunes, and eroded ventifacts (fluted and grooved rocks). Pathfinder found a dusty lower atmosphere, early morning water ice clouds, and morning near-surface air temperatures that changed abruptly with time and height. Small scale vortices, interpreted to be dust devils, were observed repeatedly in the afternoon by the meteorology instruments and have been imaged

Current—voltage curve of sodium channels and concentration dependence of sodium permeability in frog skin

1. The inward facing membranes of in vitro frog skin epithelium were depolarized with solutions of high K concentration. The electrical properties of the epithelium are then expected to be governed by the outward facing, Na-selective membrane. 2. In this state, the transepithelial voltage (V) was clamped to zero and step-changes of Na activity in the outer solution ((Na)(o)) were performed with a fast-flow chamber at constant ionic strength, while the short-circuit current was recorded. 3. At pre-selected times after a step-change of (Na)(o) the current response (I) to a fast voltage staircase was recorded. This procedure was repeated after blocking the Na channels with amiloride to obtain the current—voltage curve of transmembrane and paracellular shunt pathways. The current—voltage curve of the Na channels was computed by subtracting the shunt current from the total current. 4. The instantaneous I(Na)—V curve thus obtained at a given (Na)(o) could easily be fitted with the constant field equation in the range between -50 and zero mV. This fit yielded approximate estimates of P(Na), the Na— permeability of the Na-selective membrane (at this (Na)(o)) and the cellular Na activity, (Na)(c). As residual properties of the serosal membrane were ignored the computed values are expected to underestimate the true ones. 5. At constant (Na)(c), the steady-state value of 1/P(Na) increases linearly with (Na)(o). Error analysis and the effect of drugs show that the dependence is not due to the residual properties of the inward facing membranes but reflects the true behaviour of P(Na). 6. The steady-state P(Na) at a given (Na)(o) is smaller than the transient P(Na) observed right after a stepwise increase of (Na)(o) to this value. The time constant of P(Na)-relaxation is in the order of seconds. 7. In conclusion, Na transport through open Na-selective channels of the outward facing membrane of the stratum granulosum cells can be described as an electrodiffusion process which as such does not saturate with increasing (Na)(o). However, when added to the outer border of the membrane Na causes a decrease of P(Na) within several seconds. It is considered that binding of Na results in closure of Na channels