No abstract

Circulating concentrations of oxytocin increase to 10-40 pM in rats in response to osmotic stimuli, suggesting that oxytocin could play a role in regulation of water balance. The present studies tested whether oxytocin at such concentrations increases osmotic water permeability (Pf) in isolated perfused terminal inner medullary collecting ducts (IMCD). In IMCD segments from Sprague-Dawley rats, 20 pM oxytocin added to the peritubular bath caused a two- to threefold increase in Pf, whereas 200 pM oxytocin increased Pf by five- to sixfold (n = 8, P < 0.01). IMCD from Brattleboro rats, which manifest central diabetes insipidus, exhibited a 2.8-fold increase in Pf in response to 20 pM oxytocin and a 4.7-fold increase in response to 200 pM oxytocin. However, in Brattleboro rats, the response to 20 pM oxytocin was dependent on prior water restriction of the rats. Immunoblotting showed no change in the expression of the aquaporin-CD water channel in Brattleboro rats in response to water restriction. Nevertheless, immunofluorescence studies of inner medullary tissue from Brattleboro rats revealed a marked redistribution of the aquaporin-CD water channels to a predominantly apical and subapical localization in IMCD cells in response to water restriction, similar to the redistribution seen in response to vasopressin. Mathematical modeling studies revealed that the measured increase in Pf in response to oxytocin is sufficient to generate a concentrated urine. We conclude that oxytocin can function physiologically as an antidiuretic hormone, mimicking the short-term action of vasopressin on water permeability, albeit with somewhat lower potency.

A modified Newton-Raphson method for solving finite difference equations for the renal counterflow system is described. The method has proved generally stable and efficient, and has given significant computational results for a variety of models: calculations on single solute models of the coupled vasa recta nephron counterflow system have shown that for large water and solute permeabilities of the exchanging membranes, behavior of the non-ideal system approaches that of the previously described ideal central core model. Concentration by salt and urea mixing in two solute models has been analyzed and previous conclusions from central core models have been found to remain valid in non-ideal systems. The numerical solutions have set some order of magnitude bounds on permeability requirements for concentration in different types of non-ideal systems. Finally, from the detailed concentration profiles it has been possible to relate the rate of free energy creation and dissipation from transmembrane transport of solutes and water to the net rate of free energy efflux from the counterflow system, and so to compute in a given model the fraction of power used for solute concentration.It has been proposed in previous papers (1-3) that the behavior of the intricately coupled nephrovascular counterflow system of the renal medulla (4) approaches as a limiting case that of a four-tube model: the vascular counterflow exchanger is represented by a single tube-the central core, closed at the papillary end and open at the corticomedullary junction-which exchanges with three other tubes corresponding respectively to ascending Henle's limb (AHL), descending Henle's limb (DHL), and collecting duct (CD). Under the assumption that total solute concentrations in core, DHL, and CD are nearly the same at each level of the medulla, it has been possible to develop an approximate analytic theory of the ideal central core concentrating engine and so of the medullary counterflow system. This assumption implies very high solute and water permeability of the vasa recta and very high osmotic water permeability and (or) solute permeability of DHL and CD. The behavior of non-ideal models with finite permeabilities will deviate from that of the ideal central core model. In general, the differential equations describing non-ideal models must be solved numerically. For certain single-solute models this has been done by converting the two-point boundary value problem to an initial value problem (5), but this method tends to be unstable, requiring very good initial estimates to converge, and does not extend readily to two-solute models.In this paper we outline a modified Newton-Raphson method for solving globally finite difference equations approximating the differential equations. The method has proved generally applicable to a variety of models of the renal counterflow system. In this paper we summarize some significant preliminary computational results. Detailed descriptions of both the method and the results are in preparation. [2) [3] [4] ...

The present study was carried out to test directly whether isolated perfused rabbit cortical collecting ducts (CCDs) spontaneously generate a luminal disequilibrium pH. We determined disequilibrium pH as the difference between 1) the actual luminal pH measured by perfusing the lumen with a membrane-impermeant pH-sensitive dye [1,4-dihydroxyphthalonitrile (1,4-DHPN)] and 2) equilibrium pH calculated from the measured total CO2 concentration in fluid collected at the end of the tubule. When the peritubular bath and perfusate had the same composition, a statistically significant acidic disequilibrium pH was found (mean -0.14 units). To determine whether the disequilibrium pH is due to an absolute lack of luminal carbonic anhydrase, we measured the effective rate constant for carbonic acid dehydration in the lumen (k-1). To do this, a lumen-to-bath NH3 concentration gradient was imposed, and the luminal pH was measured along the tubule with 1,4-DHPN. NH3 absorption caused a luminal disequilibrium pH (due to dissociation of NH+4 to NH3 and H+), whose profile along the lumen is dependent on k-1 and NH3 permeability (PNH3). PNH3 and k-1 were estimated from the luminal pH profiles using a mathematical model of proton and buffer transport. The measured k-1 (37 s-1) is within the reported range of values for uncatalyzed H2CO3 dehydration. Calculations demonstrate that the measured PNH3 (2 X 10(-3) cm/s) is high enough and the measured k-1 is low enough to explain ammonia secretion rates seen in previous studies. We conclude that proton secretion in the CCD generates an acidic luminal disequilibrium pH, associated with an absolute lack of luminal carbonic anhydrase, which enhances the net rate of NH3 secretion.