A B S T R A C T To study the physical properties of renal tubular basement membranes directly, the epithelial layer of single isolated perfused rabbit proximal convoluted, proximal straight, and cortical collecting tubules was removed with sodium desoxycholate. Tubular segments were perfused using micropipets. The distal end of each segment was occluded in order to simplify the measurement of transmembrane water flow. The relation between outer tubular diameter and applied transmural pressure was identical in intact tubules and their respective isolated tubular basement membranes indicating that the basement membrane determines tubular distensibility. Young's modulus for basement membranes from all tubular segments corresponded to that of tendon collagen. Membrane hydraulic conductivity was measured in two ways: (a) from the rate of transmural flow in response to an applied difference in hydrostatic pressure and, (b) from the rate of transmural flow in response to a difference in colloid osmotic pressure. The hydraulic conductivity of tubular basement membranes was 300-800 times greater than that of the intact epithelial layer. Basement membrane hydraulic conductance was similar to that of peritubular and glomerular capillaries in vivo. The hydrostatic conductance of tubular basement membranes exceeded the osmotic conductance by 3-10-fold owing largely to the fact that the membranes were moderately permeable to the osmotic solute (albumin). In view of these findings we suggest that oncotic and hydrostatic pressure may play an important role in the movement of tubular absorbate from the epithelial compartment into the renal interstitium.

A morphometric technique is used to estimate the absolute and relative surface areas of the brush border microvilli and cell walls bordering lateral intercellular spaces. In isolated, perfused proximal tubule from rabbit, the luminal and lateral surfaces are equal in area. For proximal convoluted tubules (PCT) each surface is 4.1 +/- 0.3 mu2/mu3 of epithelial cell volume or approximately 2.9 X 10(6) mu2/mm of tubule length. In proximal straight tubules (PST) the areas are 2.6 +/- 0.2 mu2/mu3 or 1.2 X 10(6) mu2/mm. Brush border enlarges the apical cell surface 36-fold in PCT and 15-fold in PST. The luminal and lateral cell surfaces each are approximately 20-fold (in PCT) and 10-fold (in PST) greater than the areas of the basal cell surface and tubule basement membrane. These data may be important in the context of an intercellular transport model.

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The interrelationships between changes in intracellular calcium concentration ([Ca2+]i) and intracellular pH in Madin-Darby canine kidney cells and kidney glomerular epithelial cells exposed to various stimuli were analyzed simultaneously using a new design of a fluorescence video microscope. Cells were double labeled with indo 1 and SNARF 1 dyes and were excited simultaneously at 350 and 540 nm. Images at four emission wavelengths were captured simultaneously at 405, 475, 575, and 640 nm at 30 frames/s for the two ratio dyes. SNARF sensitivity to pH between 6.5 and 8.0 was unchanged by [Ca2+]i. The SNARF ratio maps were used to correct the pH-dependent changes in the calculation of local cell calcium. NH4Cl loading produced the expected alkalinization and a concurrent rise in [Ca2+]i. When the NH4Cl was removed and the cells became acidic, a second rise in [Ca2+]i was recorded. Both changes in [Ca2+]i were from intracellular stores since they persisted in the absence of extracellular calcium. The findings demonstrate the need for pH correction of indo 1 recordings.

In electron micrographs of proximal convoluted (PCT) and proximal straight tubules (PST), epithelial height was divided into five zones of equal thickness. Morphometric techniques were used to calculate surface area of cell wall bordering intercellular channels in each zone. Surface concentration of total lateral cell surface is 3.85 mu2/mu3 of PCT and 2.90 mu2/mu3 of PST. For tubules normalized to outer diameter = 40mu and inner diameter = 25mu, total lateral area is 29 X 10(5) mu2/mm of PCT and 22 X 10(5) mu2/mm of PST. Zone 5 adjacent to basement membrane has similar area (congruent to 17 X 10(5) mu2/mm) and fine structure in PCT and PST. However, the luminal four-fifths of the two cells differ markedly. Lateral area in PCT zones 1 through 4 increases approximately exponentially (from 1.1 X 10(5) to 6.4 X 10(5) mu2/mm) and constitutes 44% of total area. Respective areas in PST increase at a rate greater than exponential (from 0.7 X 10(5) to 2.6 X 10(5) mu2/mm) but constitute only 23% of total. From these data and the estimated number of cells per millimeter of tubule (825), circumferences of individual cells were estimated and quantitative three-dimensional cell models were constructed. The shape of intercellular channels is similar to that of the space between concentric, truncated and pleated horns.