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H H Ussing

Publications and source records attributed to H H Ussing.

At least 37 records · Page 2Linked to original sources

Evaluation of transport pathways for Na+ across frog skin epithelium by means of presteady-state flux ratio.

A method is described that makes it possible to separate the sodium fluxes through the isolated frog skin into sets characteristic of the cellular and the paracellular pathway, respectively. If there are two significant pathways for the ion, and if they differ with respect to flux ratio as well as mean passage time, the flux ratios for the individual pathways can be obtained from a set of inward and outward tracer fluxes, covering the time from the addition of the tracers until the achievement of constant fluxes in both directions.

Animals↗

Determination of the electromotive force of active sodium transport in frog skin epithelium (Rana temporaria) from presteady-state flux ratio experiments.

The presteady-state influxes and effluxes of sodium across frog skin epithelium have been determined as a function of time while all electrophysiological parameters were maintained constant. The fluxes measured were resolved in the fractions which have passed a pathway through the cells and those that have used a paracellular pathway. The procedure is based on the theory that all presteady-state flux ratios have to be equal to the steady-state flux ratio if only one pathway is involved. The flux ratios for the transcellular route were used to calculate the electromotive force of the sodium pump. The calculation hinges on the assumptions (a) that both influx and efflux have to pass through the sodium pump and (b) that single file diffusion of sodium is not taking place anywhere along the path. The validity of both assumptions is discussed. Our calculated values for the electromotive force of the sodium pump EaNa vary between 146 and 200 mV, which is in agreement with the energy of the ATP/ADP system. There is a distinct indication that, as the electrochemical gradient for sodium opposing the transport is being increased, the emf increases towards an asymptotic value around 200 mV. The relation between the value of EaNa and the cellular phosphorylation potential for ATP is discussed.

Animals↗

Analysis of presteady-state Na+ fluxes across the rabbit corneal endothelium.

Instead of the conventional steady-state fluxes, the presteady-state fluxes of 22Na across the rabbit corneal endothelium were measured. In contrast to reports that there is no net Na+ movement across the corneal endothelium, we find a net transport of Na+ across this tissue. The direction of net Na+ flux is from the stromal to the aqueous side and the magnitude is 2.3 +/- 0.4 mueq/cm2 X hr (n = 11, SEM). Net Na+ transport is inhibited in the presence of ouabain (10(-4) M). Acetazolamide (10(-4) M) has only a slight inhibitory effect on the rate of Na+ transport but decreases the transendothelial potential difference by about 30%. The passive component of the Na+ transport has been estimated by analyzing the presteady-state influx and efflux curves and found to occur 10% via cellular and 90% via paracellular routes. The analysis for the separation of the pathways has been based on a recently proposed theory which holds that the flux ratio, regardless of its driving forces, is independent of time.

Acetazolamide↗

Volume regulation of frog skin epithelium.

Previous results (MacRobbie & Ussing 1961) in combination with published values for cellular chloride concentration and for intracellular potentials show that the chloride concentration in frog skin epithelium cells is higher than predicted for equilibrium with the inside bathing solution. Both the apical and the basolateral membrane of these cells are normally almost tight to chloride, so that the maintenance of the high chloride concentration requires little work. A basolateral permeability to chloride is, however, activated by cell swelling, and the cells lose KCl. It is now shown that the KCl thus lost cannot be regained neither in the absence of sodium in the inside bath nor in the presence of furosemide. The volume regulation reactions are, however, independent of the composition of the outside bath. It is concluded that the recovery of KCl by the epithelium is due to a basolateral co-transport of NaCl from medium to cells, combined with return of Na to the medium via the Na-K pump. The co-transport mechanism thus restores the high chloride concentration of the cells, but seems to be virtually dormant unless the cells have lost chloride.

Animals↗

The flux ratio equation under nonstationary conditions.

The time dependent (i.e., nonstationary) unidirectional fluxes through a multilayered system consisting of sandwiched layers of arbitrary composition and exhibiting arbitrary potential and resistance profiles have been calculated, assuming that the flux is governed by the Smoluchowski equation (i.e., a flux resulting from a diffusion process superimposed upon a migration and/or a convection process, where part of the latter may arise from an active transport process). It is shown that during the building up of the concentration profile of the isotope inside the system towards the stationary value the ratio between the two oppositely directed, time-dependent unidirectional fluxes is, from the very first appearance of the isotope in the surrounding solutions, equal to the value of the stationary flux ratio. The practical implications of this result are discussed.

Biological Transport↗

Physiology of transport regulation.

The regulation of biological transport is discussed on the basis of studies on sodium transport through amphibian skin. The following types of regulation are briefly considered: 1) Hormonal regulation 2) Regulation of Na entry by apparent or real saturation of entry path by outside Na 3) Regulation of Na transport by changes in resistance to the counter ion (mostly chloride) 4) Role of cellular Na concentration which may act both by controlling the passive entry of Na and by influencing the pumping rate. 5) Dependence of Na entry upon cell volume. It is shown that a moderate osmotic swelling of ouabain-poisoned skins leads to excessive swelling of the whole epithelium when NaCl is present on the outside. This indicates that cell swelling leads to opening of the Na channels, but it also indicates coupling between the different layers of the epithelium.

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Quantitative relationship between active sodium transport, expansion of endoplasmic reticulum and specialized vacuoles ("scalloped sacs") in the outermost living cell layer of the frog skin epithelium (Rana temporaria)

When an isolated frog skin (Rana temporaria) is exposed to a hydrostatic pressure difference between inside and outside bathing solutions (inside pressure higher than outside) of 20-50 cm of H2O and if under these conditions the skin is short-circuited electrically, small "vacuoles" appear light-microscopically in the outermost living cell layer in the epithelium. The number of such "vacuoles" shows a linear dependency on the rate of active sodium transport as measured by the short-circuit current. Electron-microscopically, the "vacuoles" are interpreted as previously undescribed organelles, the "scalloped sacs" which are about 0.5 mu in diameter, with a wrinkled surface and bounded by a unit membrane. This organelle is in intimate contact with sacs and tubules of smooth endoplasmic reticulum. The observed increase in the number of scalloped sacs usually is accompanied by a significant expansion of the whole system of endoplasmic reticulum. Some of the "vacuoles" seen light-microscopically must indeed be expanded cisternae of endoplasmic reticulum. The findings are discussed in light of the possibility that the scalloped sacs and the endoplasmic reticulum may be involved in active transport of sodium ions.

Animals↗

Some morphological aspects of active sodium transport. The epithelium of the frog skin.

A method was experimentally tested which allows simultaneous morphological and bioelectrical studies of a tissue that performs active sodium transport, i.e., the isolated, surviving frog skin. In a four cell lucite chamber with four separate electric potential and current circuits, skin specimens for morphological observation (light and electron microscopy) were fixed in situ in well-defined functional states. The rate of active sodium transport through the epithelium of Rana temporaria skin was modified by changing the strength of the electric current passed through the specimens. A marked, reversible swelling of the outermost layer of the stratum granulosum was observed during short circuiting of the skin compared to the homogeneous appearance of the epithelium under open circuit conditions. Doubling the ingoing current led to an additional small increase of the swelling or the appearance of islets of cell necrosis in the same layer. There were signs of a slight shrinkage of the underlying cell layers. The observations are discussed in the light of previous bioelectrical and morphological observations.

Animals↗