A B S T R A C T To study the mechanism of phosphate reabsorption by the proximal tubule and the effect of parathyroid hormone (PTH), microperfusion experiments were carried out in rats. Segments of proximal tubule isolated by oil blocks were perfused in vivo with one of three solutions, each containing 152 meq/liter Na' and 2 mmol/liter phosphate, but otherwise differing in composition. The pH of solution 1 was 6.05-6.63, indicating that 60-85% of the phosphate was in the form of H2PO4. The pH of solution 2 was 7.56-7.85, and 85-92% of the phosphate was in the form of HP04=. Solution 3 contained HC03-and glucose and had a pH of 7.50-7.65. When the proximal tubules were perfused with solution 1, the "2P concentration in the collected perfusate was found to be consistently lower than in the initial perfusion solution. In sharp contrast, when the tubules were perfused with solutions 2 or 3, s'P concentration usually rose above that in the initial solution. Water (and presumably Na') reabsorption, as measured with ['H]inulin, was the same with the acid and alkaline solutions. Administration of partially purified PTH clearly prevented the fall in phosphate concentration with the acid solution, but had a less discernible effect on phosphate reabsorption with the two alkaline solutions. Measurements of pH within the perfused segments with antimony microelectrodes demonstrated that PTH enhanced alkalinization of the acid perfusion solution. The findings are consistent with the view that H2POf is reabsorbed preferentially over HPO4=. This can be attributed to either an active transport mechanism for H2PO4J or selective membrane permeability to this anion. PTH appears to either inhibit an active transport process for H2PO4-, or to interfere with pas-

Controversial evidence exists as to whether electroneutral Cl-OH exchange occurs across the proximal tubular brush border membrane of the mammalian kidney. To help resolve this controversy, we carried out a series of experiments using rabbit renal cortical brush border membrane vesicles to determine the effect of changes in pH on 36Cl uptake across this membrane. Reducing pH equally inside and outside of the vesicle stimulated Cl-Cl exchange as well as chloride uptake. A proton gradient (out greater than in) stimulated and caused an overshoot in unidirectional Cl uptake. All three of these processes were inhibited by 4-acetoamido-4-isothiocyano-2,2-disulfonic stilbene. Valinomycin with K+ out-in added to shunt any proton diffusion potential, minimally inhibited pH gradient-dependent Cl uptake, and carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone (FCCP), added to increase the proton diffusion potential, minimally stimulated pH gradient-dependent Cl uptake. Sodium-glucose cotransport (an electrogenic system) was used to assess the efficacy of these ionophores. FCCP markedly enhanced and valinomycin markedly inhibited pH gradient stimulated sodium-glucose cotransport. Valinomycin added to vesicles with a 180/0 meq K+ out greater than in gradient stimulated less Cl uptake than a 6.0/7.5 pH out greater than in gradient. We conclude that these results provide strong evidence for the existence of an electroneutral Cl-OH exchanger or Cl-H symporter in this membrane.

Silicone rubber injections of methyl salicylate-cleared rat kidneys were performed. In 50 of 56 injections of superficial nephrons with their accompanying blood supply, the efferent vessel and early proximal tubule were closely approximated. In 18 of 21 tubular injections filling through the pars recta, the proximal tubule folded upon itself with early and late proximal segments, in close contact, located over their parent glomerulus, and the midproximal segments separate and located over their parent interlobular artery. The distribution of blood was serially through the early-late proximal region above the glomerulus via a long unbranched efferent vessel, via branches over the capsular surface, via capillaries down through the midproximal region, then into the interlobular vein. The observed anatomical pattern of the superficial nephron appears to permit direct functional interactions between the juxtaposed early and late proximal tubule, and in turn may effect midproximal function via the distribution of blood (modified by early proximal) from the efferent vessel to midproximal convolutions. In addition, the relationship between specific segments of the proximal tubule and specific portions of the postglomerular peritubular blood supply may be important in determining the distribution of peritubular physical forces to these nephrons.

The majority of the oxygen consumed by the rat kidney appears to occur in the proximal tubule. Therefore changes in metabolically linked ion transport in this segment of the nephron should result in changes in renal oxygen consumption. To study the role of bicarbonate reabsorption in metabolically linked proximal tubular ion transport a series of micropuncture-clearance-extraction experiments were performed comparing the effects of the carbonic anhydrase inhibitor benzolamide and of hypertonic sodium bicarbonate infusion with control conditions in the rat. End-proximal tubular fluid and chloride reabsorption were measured. From these, the rates of sodium and bicarbonate reabsorption were estimated. Simultaneously with the tubular fluids, extraction collections were obtained for determination of renal oxygen consumption. Both benzolamide and hypertonic bicarbonate reduced proximal tubular fluid reabsorption while concomitantly reducing the transepithelial gradient for chloride. The mean rate of renal oxygen consumption did not differ from the control rate in either experimental group and could be dissociated from the calculated net rates of proximal tubular sodium, chloride, and bicarbonate reabsorption. We interpret these data as evidence that proximal tubular hydrogen ion secretion supporting bicarbonate reabsorption requires at most small amounts of oxidative energy, less than detectable by these techniques. The data, in contrast, support the conclusion that the chloride-bicarbonate transepithelial gradient appears to be an important passive driving force in vivo for proximal tubular fluid reabsorption.

We administered an organic mercurial diuretic compound, an organic mercurial non-diuretic compound and an inhibitor of carbonic anhydrase to anesthetized dogs. Using conventional clearance methods, we measured renal plasma flow, filtration rate, sodium reabsorption and oxygen consumption. The mercurial diuretic compound blocked a fraction of both sodium reabsorption and oxygen consumption. The non-diuretic compound affected neither sodium reabsorption nor oxygen consumption. The inhibitor of carbonic anhydrase blocked the reabsorption of a fraction of the filtered sodium without affecting oxygen consumption. We conclude that (1) sodium chloride reabsorption is dependent upon oxidative metabolism for energy; (2) binding of renal cortical sulfhydryl compounds affects neither sodium transport nor oxidative metabolism; (3) the mechanism for sodium reabsorption by ion exchange is not dependent upon oxidative metabolism for energy.