Biomedical subjects
Publications and source records attributed to K R Spring.
The lateral intercellular spaces of Necturus gallbladder epithelium were seen and measured while the living tissue was perfused in a new chamber. The compliance of the lateral cell membranes was calculated from the measured pressure-volume characteristics of the lateral intercellular spaces.
Explore the source record for details and available documents.
Necturus proximal tubule lumen was filled with solutions of Na36Cl or [36Cl]tetramethylammonium ([36Cl]TMA) and the tracer disappearance was measured. With these tracers it was possible to differentiate between chloride fluxes across the cellular and the extracellular shunt pathways. Since it was previously shown that chloride does not enter tubule cells from the lumen unless Na is also present in the lumen, the [36Cl]TMA disappearance rate gave the shunt flux of chloride while the Na36Cl disappearance rate gave the sum of the transcellular and the shunt fluxes. The transcellular tracer chloride flux was unaffected by changes in the transepithelial potential difference, and the rate constant for the chloride flux from lumen to cell was identical to that previously reported for luminal sodium entry. These observations support the conclusion that a coupled transport of NaCl, in an electrically silent form, occurs across the luminal membrane of the Necturus proximal tubule cell. Shunt chloride flux was directly proportional to the electrical driving force, indicating diffusional chloride movement out of the lumen into the shunt pathway.
The dependence of proximal tubular sodium and fluid readsorption on the Na(+) concentration of the luminal and peritubular fluid was studied in the perfused necturus kidney. Fluid droplets, separated by oil from the tubular contents and identical in composition to the vascular perfusate, were introduced into proximal tubules, reaspirated, and analyzed for Na(+) and [(14)C]mannitol. In addition, fluid transport was measured in short-circuited fluid samples by observing the rate of change in length of the split droplets in the tubular lumen. Both reabsorptive fluid and calculated Na fluxes were simple, storable functions of the perfusate Na(+) concentration (K(m) = 35-39 mM/liter, V(max) = 1.37 control value). Intracellular Na(+), determined by tissue analysis, and open-circuit transepithelial electrical potential differences were also saturable functions of extracellular Na(+). In contrast, net reabsorptive fluid and Na(+) fluxes were linearly dependent on intracellular Na(+) and showed no saturation, even at sharply elevated cellular sodium concentrations. These concentrations were achieved by addition of amphotericin B to the luminal perfusate, a maneuver which increased the rate of Na(+) entry into the tubule cells and caused a proportionate rise in net Na(+) flux. It is concluded that active peritubular sodium transport in proximal tubule cells of necturus is normally unsaturated and remains so even after amphotericin-induced enhancement of luminal Na(+) entry. Transepithelial movement of NaCl may be described by a model with a saturable luminal entry step of Na(+) or NaCl into the cell and a second, unsaturated active transport step of Na(+) across the peritubular cell boundary.
Steady-state bidirectional sodium fluxes were measured across Necturus proximal tubules. New methods for capillary perfusion and collection of venous effluent enabled flux determination to be made from the appearance of luminal tracer in the capillaries. Fluxes and permeability were measured in the absence of net fluid reabsorption. The sodium permeability measured in the plasma-to-lumen direction was 3 X 10(-6) cm/s. The flux ratio (lumen-to-plasma/plasma-to-lumen) was about twice the passive value calculated from the measured concentrations and potentials. Estimates for the permeability and flux across the shunt pathway were obtained from nonsteady-state flux determinations. The shunt pathway appeared to be the most significant route for passive sodium movement from plasma-to-lumen. Nonsteady-state tracer measurements also enabled an estimate to be made of the lumenal cell membrane permeability and unidirectional sodium flux. Two-thirds of the lumen-to-plasma flux was calculated to traverse the cellular path and the remainder through the shunt. Approximately one-third of the intracellular sodium was found to exchange rapidly with tracer.