bluuebunny/crossref_metadata_2025 huggingface dataset:
Amino acid content of rat renal cortex and the response to in vitro incubation
Authors: Blazer-Yost B. , Reynolds R. , Segal S.
Published: December 2017

The concentration of aspartic acid, threonine, serine, glycine, and alanine is significantly higher in newborn rat renal cortex than in the adult tissue, while phenylalanine and histidine are higher in the adult. When adult cortical slices are placed in bicarbonate buffer at room temperature for 20 min there is a 30-60% decrease in the levels of all amino acids except for lysine, which is slightly higher, and methionine and serine, which do not change. Under the same conditions, newborn cortical slices reveal a similar decrease in only glycine, tyrosine, histidine, and the branched-chain amino acids. On subsequent in vitro incubation of the cortical slices at 37 degrees C for 120 min the concentrations in adult tissue remain at the lower values observed on removal from buffer at room temperature except that glutamic acid, glycine, and lysine levels decrease further and serine increases to the concentration found in fresh tissue. Newborn tissue when incubated at 37 degrees C for 120 min shows amino acid concentrations comparable to unincubated fresh tissue for all except aspartic acid, glutamic acid, serine, and phenylalanine, which reach levels higher than unincubated tissue. The ability of newborn tissue to maintain amino acid pools may play a role in the enhanced transport of some amino acids resulting from preincubation at 37 degrees C (Reynolds et al. Science 184: 68-69, 1974; Reynolds and Segal, Biochim. Biophys. Acta 406: 513-525, 1976).
Sodium in renal tubular acidification kinetics
Authors: de Mello Aires M. , Malnic G.
Published: December 2017

Renal proximal tubules and their peritubular capillaries were perfused with mammalian Ringer solutions containing different sodium concentrations. In stop-flow microperfusion experiments, the pH was measured by means of antimony microelectrodes, permitting calculation of rates of H ion secretion and bicarbonate reabsorption. These rates, as well as transepithelial pH and bicarbonate gradients, were significantly reduced at ambient concentrations of 20 and 4 meq/liter Na+. However, even at the lowest sodium concentrations (4 meq/liter), H ion secretion was still 74%, and bicarbonate reabsorption of 64% of control rates. In similar conditions, sodium reabsorption as measured by the split-droplet technique fell to practically zero. Ouabain, 10(-3) M, in capillaries reduced bicarbonate reabsorption by 31%, and 3 X 10(-4) M furosemide in lumen and capillaries reduced acidification by 29%. At pH 8--9 in capillaries, sodium transport was normal while acidification was markedly reduced. These data show that low sodium levels impair renal tubular acidification, but they do not support a rigid coupling of these transport processes.
Urinary concentrating mechanism in the desert rodent Psammomys obesus
Authors: Jamison R. L. , Roinel N. , de Rouffignac C.
Published: December 2017

The mechanism by which osmolality rises in fluid in the juxtamedullary descending limb was examined in Psammomys obesus by micropuncture of the left renal papilla. The concentration of Na, Cl, K, Mg, and phosphorus was determined by electron probe; Cl concentration by both probe and the microcoulometric technique of Ramsay et al. (J. Exp. Biol. 32: 822--829, 1955). The mean urine-to-plasma ultrafiltrate (U/P) osmolality was 5.5 +/- 0.46 (SE). In 43 samples obtained from the end-descending limb in 16 animals, mean values were: tubule fluid-to-plasma ultrafiltrate (TF/P) osmolality, 3.64 +/- 0.30; (TF/P)In, 6.96 +/- 0.52; (TF/P)Na, 3.53 +/- 0.27; (TF/P)Cl-e, 4.11 +/- 0.37 (-e is electron probe determination); and (TF/P)Cl-r, 4.25 +/- 0.37 (-r is determination by method of Ramsay). In terms of percentage of filtered ion remaining at the end-descending limb, Na was 54 +/- 3.7%, Cl-e was 61 +/- 3.6%, and Cl-r was 63 +/- 4.0%. The percentage of filtered chloride remaining averaged 80% or more in three animals, equalled or slightly exceeded 100% in six individual descending limb samples, and in all animals was a function of loop fluid osmolality (y = 8.61x + 31.9, r = 0.66, P less than 0.01). The results are most consistent with the thesis that both water extraction and transepithelial NaCl addition contribute to the rise in osmolality of fluid in the descending limb of the Psammomys.
Uric acid transport in brush border membrane vesicles isolated from rabbit kidney
Authors: Boumendil-Podevin E. F. , Podevin R. A. , Priol C.
Published: December 2017

The transport of uric acid was studied in brush border membrane vesicles isolated from rabbit kidney. The uptake of uric acid by the vesicles was osmotically sensitive and occurred in the absence of significant uric acid degradation. Under the conditions used to evaluate transport, urate binding to the membranes represented only 10––15% of the total uptake. The initial rate of uptake was linear over the concentration range 0.04––8 mM urate. Uptake of urage was Na+ gradient independent. It was dependent on external pH and temperature with Q10 near 3. The urate uptake was inhibited reversibly by p-chloromercuribenzoate. Probenecid, ouabain, cyclic adenosine 3',5'--monophosphate, and its dibutyryl derivative had no appreciable effects. Pyrazinoic acid and pyrazinamide stimulated urate uptake. Experiments performed with osmotically shocked vesicles demonstrated that this stimulatory effect resulted from increased binding of urate to the membranes. These results indicate that in several ways urate transport in vesicles resembles that observed with more physiologically intact preparations.
Effects of the oral converting enzyme inhibitor SQ 14225 in experimental high output failure
Authors: Williams G. M. , Davis J. O. , Freeman R. H. , DeForrest J. M. , Seymour A. A. , Rowe B. P.
Published: December 2017

The relation of the renin-angiotensin-aldosterone system to sodium retention was studied in dogs with an aortic-caval fistula and high output failure by administering orally the new converting enzyme inhibitor SQ 14225. The acute response to an initial oral dose of SQ 14225 (10 mg/kg) consisted in a fall in arterial pressure (BP) from 97 to 67 mmHg, plasma aldosterone concentration (PAC) from 21.7 to 11.3 ng/100 ml, creatinine clearance (CCr) from 89 to 44 ml/min, and filtration fraction (FF) from 39 to 18%, whereas plasma renin activity (PRA) increases from 12.7 to 36.1 ng angiotensin I.ml-1.h-1 and renal sodium excretion was unchanged. It is suggested that the marked fall in BP and CCr offset the drop in PAC and FF which favored a natriuresis. The daily responses to SQ 14225 for 3 days also showed a fall in BP and PAC but sodium excretion increased. In the animal with the best response, a striking natriuresis occurred. These findings demonstrate an important role for the renin-angiotensin-aldosterone system in experimental high output failure.
Renal metabolism of N6,O2'-dibutyryl adenosine 3',5'-monophosphate
Authors: Coulson R. , Harrington W. W.
Published: December 2017

Metabolism of dibutyryl cyclic AMP was studied by including the 3H- or C-labeled nucleotide (0.1 mM, 5 mumol) in the recirculating perfusate of the isolated rat kidney. Kidneys were perfused with nucleotide for 60 min. Dibutyryl cyclic AMP was almost completely cleared from the perfusate, about one-quarter as urinary excretion principally by probenecid-sensitive secretion and about one-half as metabolism beyond 3'-phosphate bond cleavage. The principal metabolite, N6-monobutyryl adenosine, accounted for one-third of added dibutyryl cyclic AMP. The remaining metabolites were ATP, ADP AMP, and N6-monobutyryl AMP. Dibutyryl cyclic AMP (0.1 or 1.0 mM) elevated renal ATP but did not alter uricogenesis. Both dibutyryl cyclic AMP and cyclic AMP at 0.2 mM produced similar activation and subcellular redistribution of renal protein kinase. N6-monobutyryl adenosine, unlike adenosine, had no effect on the renal activity of adenylate cyclase, low Km cyclic AMP phosphodiesterase, and protein kinase. Dibutyryl cyclic AMP is like exogenous cyclic AMP in that it penetrates the rat kidney, activates protein kinase, and is metabolized to ATP (R. Coulson, J. Biol. Chem. 251: 4958-4967, 1976), but is unlike cyclic AMP in its extent of secretion and metabolism to ATP and urate and in its formation of the unique metabolites N6-monobutyryl AMP and N6-monobutyryl adenosine.
Amphotericin B and K+ transport across excised toad urinary bladder
Authors: Gatzy J. T. , Reuss L. , Finn A. L.
Published: December 2017

The transmural electric PD of bladders bathed by Na2SO4 Ringer was not affected by amphotericin (5 x 10(-6) M, mucosal) but the PD followed the direction for K+ diffusion in the presence of a transmural K+ gradient. Increases in bathing solution K+ increased conductance. Ouabain pretreatment did not affect drug-induced changes in PD or conductance. Unidirectional fluxes of radiolabeled Na+ and K+ but not SO42- across the short-circuited bladder were increased by amphotericin. Ninety percent of the rise in the serosal-to-mucosal flow of Na+ disappeared when mucosal Na+ was replaced by choline. Amphotericin induced a 20-fold increase in mucosal-to-serosal K+ flux but K+ serosal-to-mucosal flow increased 200-fold. This flux asymmetry persisted for 110 min, was abolished by pre- or posttreatment with ouabain, and was immeasurable when bathing solution K+ was increased from 2.4 to 59 meq/liter. With 2.4 meq K+/liter the ratio of active Na+ reabsorption to K+ secretion was 8 to 1, but K+ secretion was not closely linked to Na+ transport. The results suggest that amphotericin induces a paracellular K+-selective path, Na+ isotope exchange, and K+ secretion.
Effect of sodium restriction on plasma renin activity and renin granules in rat kidney
Authors: Morimoto S. , Abe R. , Fukuhara A. , Tanaka K. , Yamamoto K.
Published: December 2017

The present study was carried out to procure detailed information on the relationship between chronic sodium restriction and renin content of kidneys at a subcellular level in the rat. Renin granules (RG) were separated by a discontinuous sucrose-density gradient (from 1.2 to 1.7 M) centrifugation. In control rats, RG were mainly recovered in the fractions corresponding to 1.5 M sucrose, whereas most of the mitochondria, lysosomes, and microsomes equilibrated in upper fractions. The RG fraction contained approximately 60% of total granular renin activity. Low sodium intake for 4 wk resulted in a 12.4-fold increase in plasma renin activity and led to a 2.6-fold increase in renin activity of the RG fraction. But in sodium-restricted rats there was no alteration in the distribution pattern of renin activity on sucrose-density gradients, indicating that there was no change in the density of RG. These results provide evidence for increased renin activity in storage granules following chronic sodium restriction.
Volume expansion-induced alterations in proximal tubule chloride gradient in the rat
Authors: Galla J. H. , Luke R. G.
Published: December 2017

To elucidate the mechanisms by which acute volume expansion (AVE) induces a decrease in proximal tubule transepithelial chloride gradient, male Sprague-Dawley rats were studied before and after AVE with Ringer lactate. In group 1, after AVE equivalent to 10% body wt, there were decreases in both tubule fluid to plasma inulin ratio ((TF/P)In) (from 2.28 +/- 0.10 to 1.57 +/- 0.05) and tubule fluid to ultrafiltrate chloride ratio ((TF/UF)Cl) (from 1.25 +/- 0.02 to 1.18 +/- 0.02). Group 2 was studied during carbonic anhydrase inhibition (CAI) produced by benzolamide before and during superimposed AVE (20% body wr). Both (TF/P)In (from 1.91 +/- 0.10 to 1.41 +/- 0.08) and (TF/UF)Cl (from 1.07 +/- 0.02 to 1.01+/- 0.01) decreased. Group 3 was studied during maintained AVE (15% body wt) as a control for group 4, in which CAI was superimposed on maintained AVE. In group 3, (TF/P)In and (TF/UF)Cl did not change, but in group 4 CAI was associated with a decrease in (TF/P)In (from 1.55 +/- 0.05 to 1.21 +/- 0.05) and in (TF/UF)Cl (from 1.16 +/- 0.01 to 1.04 +/- 0.07). These data suggest that in the superficial proximal convoluted tubule of the rat, AVE-induced alterations in transepithelial chloride gradient are dependent on a mechanism(s) other than changes in carbonic anhydrase-mediated bicarbonate reabsorption.
A network thermodynamic model of salt and water flow across the kidney proximal tubule
Authors: Thomas S. R. , Mikulecky D. C.
Published: December 2017

Page F638: S. R. Thomas and D. C. Mikulecky. “A network thermodynamic model of salt and water flow across the kidney proximal tubule.” Page F640: In equation 4, (c12 – 2c1c2 + c22) should be (c22 – c12)/2. Page 646: In the program listing of Fig. 4 the line 00320 RJS3 1 6 2.457E4 should read instead 00320 RJS3 11 16 2.457E4.
Renal handling of phenol red. IV. Tubular localization in rabbit and rat kidney in vivo
Authors: Sheikh M. I. , Moller J. V.
Published: December 2017

In the rabbit kidney accumulation of phenol red in cortex tissue is directly related to phenol red excretion. In histological preparations of rabbit cortex the major part of phenol red is localized to the pars recta of proximal tubules at low plasma concentrations of dye. The extra uptake of dye by the pars recta is abolished by administration of a high dose of probenecid and also by high plasma dye concentrations, when dye secretion is low relative to tubular reabsorption. Tissue accumulation of phenol red in the rat exhibits features similar to those in the rabbit. However, extra dye uptake in the pars recta is maintained after administration of probenecid, and disappears after intravenous injection of phenol red during ureteral occlusion to impede access of dye from tubule fluid to the luminal membrane. It is concluded that in the rabbit phenol red uptake by the pars recta probably is due to tubular secretion across the peritubular membrane, whereas in the rat extra uptake of dye by this segment is consistent with uptake at the luminal cell membrane.
Lactate absorption in Thamnophis proximal tubule: transport versus metabolism
Authors: Brand P. H. , Stansbury R. S.
Published: December 2017

Proximal tubules from the kidney of Thamnophis (garter snake) were perfused in vitro and unidirectional fluxes of lactate measured using L(+)-[U-14C]lactate, (lactate concentration, 1 mM). The lumen-to-bath (absorptive) flux (Jlb lact) significantly exceeded the bath-to-lumen flux (backflux) (Jbl lact) in each of 12 tubules (seven distal proximal and five proximal proximal). The flux ratio (Jlb lact/Jbl lact) was approximately 3.00. At flow rates of 13-16 nl/min and lactate concentration of 1 mM the net flux was about 1.60 pmol . min-1 . mm-1 in both proximal proximal and distal proximal segments. Both fluxes were decreased by perfusion at 5 degrees C. To determin e the contribution of metabolism of lactate to its absorption, Jlb lact was measured at 25 degrees C in 10 distal proximal tubules during perfusion with [14C]lactate, lactate concentration, 1 mM, and with [methoxy-3H]inulin. In these experiments, the amount of 14C found in the bath was 93% of the amount of 14C absorbed from the lumen. Chromatography showed that all of the 14C found in the bath was [14C]lactate. These data establish that in Thamnophis proximal tubule lactate absorption occurs against an electro chemical gradient by transport of the intact lactate molecule without significant metabolism.
Competition between sodium reabsorption and gluconeogenesis in kidneys of steroid-treated rats
Authors: Silva P. , Ross B. , Spokes K.
Published: December 2017

Kidneys of rats treated with methylprednisolone show altered substrate requirements for sodium reabsorption when perfused in vitro. Such kidneys synthesize glucose from lactate at twice the rate of control. Optimum sodium reabsorption is not seen with glucose, which is normally the preferred substrate. Sodium reabsorption is restored toward normal by the combination of glucose and butyrate, by pyruvate, or by 3-mercaptopicolinate. All of these results point to a metabolic adaptation in the kidney; butyrate may improve sodium reabsorption by sparing glucose, pyruvate is a gluconeogenic precursor and an effective fuel of respiration, and 3-mercaptopicolinate is an inhibitor of gluconeogenesis. In kidneys from rats treated with methylprednisolone there is an increased requirement for metabolic energy because of the increased rate of gluconeogenesis. It is suggested that the availability of energy from glucose oxidation is limited in part by the diversion of pyruvate back to glucose. Under these special circumstances, gluconeogenesis competes with sodium reabsorption in the intact kidney.
Ischemia and tubule obstruction during acute renal failure in dogs: mannitol in protection
Authors: Burke T. J. , Cronin R. E. , Duchin K. L. , Peterson L. N. , Schrier R. W.
Published: December 2017

The pathogenetic factors involved in norepinephrine- (NE) induced reversible acute renal failure (ARF) were examined in untreated (U) and mannitol-treated (M) animals. At 3 and 24 h after NE infusion renal blood flow (RBF) was significantly higher in M compared to U animals (174 vs. 138 and 191 vs. 148 ml/min, respectively, both P less than 0.05). At 3 h, glomerular filtration rate (GFR) was higher in M animals (8 vs. 4 ml/min, P less than 0.01), while at 24 h protection was even greater (18 vs. 3 ml/min, P less than 0.01). In U animals proximal tubule pressure (Pt) was lower at 1 h than before NE (13 vs. 23 mmHg, P less than 0.01); from 1 to 3 h Pt increased to elevated levels in parallel with restoration of RBF (r = 0.62, P less than 0.01). At 3 h in U animals stop-flow pressure (SFP), as an index of glomerular capillary pressure, was below normal (35 vs. 44 mmHg, P less than 0.05) yet Pt was increased (35 vs. 23 mmHg, P less than 0.05). Thus little transglomerular pressure gradient was present for ultrafiltration. Further evidence of tubular obstruction was obtained by microperfusion at 6 nl/min, which increased Pt from 30 to 45 mmHg (P less than 0.001), a finding not present in unobstructed tubules. Delayed excretion (approximately 20 min) of microinjected [3H]inulin also was compatible with renal ischemia and tubule obstruction. Microinjection studies provided no evidence for backleak of tubular fluid. At 1 h, Pt was higher in M vs. U animals (31 vs. 13 mmHg, P less than 0.05). In M animals at 3 h SFP was normal (50 vs. 44 mmHg) and Pt was below SFP (32 vs. 50 mmHg, P less than 0.01), thus preserving a substantial transglomerular pressure gradient for ultrafiltration. In summary, reduced GFR in U animals is characterized by a combination of reduced glomerular capillary pressure and tubule obstruction. In contrast, animals receiving mannitol were protected against ARF through maintenance of glomerular capillary pressure and prevention of tubular obstruction, perhaps by increasing Pt within the first hour of the NE insult.
Membrane mechanisms for transepithelial amino acid absorption and secretion
Authors: Schafer J. A. , Barfuss D. W.
Published: December 2017

Although amino acids are normally almost completely absorbed by the proximal nephron, specific amino acids may also exhibit active secretion under the appropriate circumstances. This brief review examines details of the influx and efflux transport processes across both lumina and basolateral membranes of the proximal tubule and through the paracellular pathway as determinants of the direction and rate of transepithelial amino acid transport. Net absorption depends upon active uptake from the lumen into tubular cells by cotransport with Na+. At least some proximal tubule cells also actively accumulate amino acids from their peritubular environment by active transport across basolateral membranes. The net direction of transepithelial movement depends on the relative rates of passive amino acid exit from the cell across luminal and basolateral membranes. Since exit across the basolateral membrane appears to be facilitated whereas luminal exit is not, normally net absorption occurs. The minimum luminal amino acid concentration and excretion rate are also dependent on a relatively low paracellular amino acid permeability, especially in the latest segments of the proximal nephron where the transepithelial concentration gradient is the greatest.
Phosphate transport by isolated rabbit cortical collecting tubule
Authors: Peraino R. A. , Suki W. N.
Published: December 2017

Renal handling of phosphate occurs in the proximal convoluted tubule. Absorption of this anion also occurs in the pars recta and distal convoluted tubule, the latter a structurally and functionally diverse segment. The purpose of the present investigation was to examine phosphate transport by the cortical collecting duct of the rabbit. Segments of cortical collecting tubule, beyond the last cortical branch, were isolated and perfused in vitro with an artificial solution simulating plasma as the perfusing and bathing medium. The perfusion solution contained either 3 or 25 mM bicarbonate. Net phosphate transport was measured using 32P as the radionuclide tracer, with identical specific activity in perfusing and bathing solutions. A net absorptive flux for phosphate was demonstrated, amounting to 2-3% of the delivered load. In addition, this absorptive flux was linearly related to perfusion rate and, thus, delivered load, but independent of the lumen bicarbonate concentration or pH.
Analysis of beta-adrenoceptors mediating renin release produced by isoproterenol in conscious dogs
Authors: Himori N. , Izumi A. , Ishimori T.
Published: December 2017

Types of beta-adrenoceptors mediating renin release induced by isoproterenol were investigated in conscious dogs. The nonselective beta-adrenoceptor blocking drugs propranolol, D-32, and pindolol significantly inhibited increases in heart rate and plasma renin activity and a fall of blood pressure produced by intravenous infusion of isoproterenol (10 microgram . kg-1 . 20 min-1). d-Propranolol and d-D-32 did not inhibit these three responses to isoproterenol. The selective beta 1-adrenoceptor blocking drug atenolol, at the oral dose of 6 mg/kg, which selectively suppressed isoproterenol-induced tachycardia, significantly inhibited the renin release caused by isoproterenol. By contrast, the renin release induced by isoproterenol was not modified by the selective beta 2-adrenoceptor blocking drug IPS-339 at an oral dose of 3 mg/kg, which fully and selectively antagonized the fall of blood pressure in response to isoproterenol. There was good correlation between suppression of isoproterenol-induced renin release and that of isoproterenol-induced tachycardia after various beta-adrenoceptor blocking drugs. These results lead to the conclusion that in conscious dogs the beta-adrenoceptors mediating release are mainly of the beta 1 type.
Relation of Na+ reabsorption to utilization of O2 and lactate in the perfused rat kidney
Authors: Cohen J. J. , Merkens L. S. , Peterson O. W.
Published: December 2017

When the ratio delta TNa+/delta QO2 is used to estimate the energy requirements for net Na+ reabsorption (TNa+), it is assumed that the entire change in renal O2 uptake (delta QO2) is utilized only for the delta TNa+. However, if increases in renal synthetic work also occur when TNa+ is increased, the energy cost for TNa+ will be overestimated. We perfused the substrate-limited isolated rat kidney at 38 degrees C, pH 7.4, a mean arterial pressure of 120 mmHg, and mean lactate concentrations between 0 and 8.3 mM. We measured QO2, TNa+, net reabsorption of lactate (Tlac), net utilization of lactate (Qlac), lactate decarboxylation rate (Qlacox), as well as the net entry rate of lactate into biosynthetic pathways (Qxslac). When no exogenous substrate was present (rates are means, g wet wt-1 . min-1) GFR was 351 +/- 38 microliter, %TNa+ was 54 +/- 2%, and QO2 was 2.85 +/- 0.31 mumol; there was also a loss of about 20% of renal tissue K+ content. When mean [lactate] greater than or equal to 0.73 mM, the loss of tissue K+ was completely prevented and %TNa+ increased to and remained at about 85%. At mean [lactate] of 8.3 mM, Tlac was 5.1 +/- 0.6 mumol, QO2 was 6.12 +/- 1.24 mumol, and GFR was 709 +/- 83 microliter. Qlac, delta Qlacox and delta TNa+ increased in parallel with each other and approaches maximal rates as [lactate] was raised. By contrast, Tlac increased as a linear function of perfusate [lactate] and was not related to changes in Qlac. The molar increases in TNa+ were 10- to 20-fold greater than the increases in Tlac. It is more probable, therefore, that lactate enhances TNa+ by providing energy from its oxidation rather than by a co-transport phenomenon. At all concentrations of lactate, more lactate was utilized (Km = 1.2 mM; Vmax = 3.4) than was decarboxylated (Km = 1.6 mM; Vmax = 1.7), indicating that as lactate concentration increased, both the synthesis of new products from lactate and Na+ reabsorption increased. We conclude that the ratio delta TNa+/delta QO2, overestimates the energy cost of Na+ reabsorption. In order to obtain an accurate estimate of the energy requirements for TNa+ in kidney, the simultaneous changes in the rate of net biosynthetic work must also be quantified as TNa+ is changed.
Hydrogen transport in rabbit kidney proximal tubules--Na:H exchange
Authors: Bichara M. , Paillard M. , Leviel F. , Gardin J. P.
Published: December 2017

In order to examine the cellular mechanisms of H transfer, we studied the [14C]DMO-derived cell pH in rabbit separated renal proximal tubules. Under normal conditions, cell pH (7.508 +/- 0.008, SE) was found to be more alkaline than medium pH (7.404 +/- 0.009, SE). The [14C]DMO-derived cell pH was not affected either by 10(-2) M unlabeled DMO or 10(-3) M probenecid. Removal of external sodium resulted in cell acidification. Addition or 10(-3) M ouabain to the medium dissipated the transmembrane sodium gradient [Na]o/[Na]i and lowered cell pH. However, the DMO-derived cell pH observed in the presence of ouabain was higher than the calculated cell pH, assuming passive distribution of H ions across the cell membrane. In the presence of ouabain, reducing [Na]o/[Na]i led to cell acidification, and augmenting [Na]o/[Na]i caused cell alkalinization. These results indicate that in the proximal tubule: 1) the H transfer from cell to lumen is active, whereas the HCO3 transfer across the basal membrane may be passive; 2) there is evidence for a Na:H exchange via a countertransport mechanism; and 3) Na:H exchange may not be the only mechanism responsible for active H transfer.
Interactions of temperature and ADH on transport processes in cortical collecting tubules
Authors: Hebert S. C. , Andreoli T. E.
Published: December 2017

We evaluated the temperature dependence of the permeability coefficients for ADH-independent and ADH-dependent zero volume flow diffusion of THO (PDw, micron/s), for ADH-dependent zero volume flow diffusion of the highly lipophilic solute n-[3H]butanol (PDb, micron/s), and for ADH-dependent lumen-to-bath osmosis (Pf, micron/s) in rabbit isolated cortical collecting tubules. The ADH-dependnet Pf and PDw data are consistent with the hypothesis that water crosses the apical plasma membranes of these tubules through narrow aqueous channels by single-file diffusion; and using the raw temperature-dependent data, we calculate that these channels contain congruent to six H2O molecules per channel, both in the presence and in the absence of ADH. Apparent activation energies (EA, kcal/mol), as an upper estimate of the true activation energies, were calculated for these transport processes. The EA for osmosis for 6–23 degrees C, 9.7 +/- 1.1 kcal/mol, was indistinguishable from that reported previously for 23–37 degrees C either in the presence of absence of ADH, and from the ADH-independent EA for THO diffusion, 10.11 +/- 0.68 kcal/mol, measured in the range 15–37 degrees C. We interpret these data to indicate that in the absence of ADH THO diffusion at zero volume flow is hindered primarily by the same sites, narrow channels in apical membranes. The apparent EA for ADH-dependent zero volume flow THO diffusion, 5.11 +/- 0.40 kcal/mol, measured over the temperature range 6–38 degrees C, is approximately half the apparent EA for osmosis in the presence or absence of ADH and ADH-independent THO diffusion. The former value, when corrected for diffusion constraints in series with apical plasma membranes, becomes indistinguishable from the latter values. These date are consistent with the possibility that two factors contribute to the ADH-mediated disparity between Pf and PDw in these tubules: cytosolic diffusion constraints that impede ADH-dependent THO diffusion but not osmotic volume flow, and narrow aqueous channels as the primary route for water flux through apical plasma membranes. Finally, the EA for n-butanol diffusion shows a break in the Arrhenius plot. Above 23 degrees C, the EA is 4.77 +/- 0.77 kcal/mol and n-butanol diffusion is impeded primarily by the cytosol. Below 23 degrees C, the EA rises to approximately 9 kcal/mol because the hydrophobic regions of apical plasma membranes contribute an increasing fraction of the total resistance to n-butanol diffusion.
Calcium and magnesium transport along the inner medullary collecting duct of the rat
Authors: Bengele H. H. , Alexander E. A. , Lechene C. P.
Published: December 2017

We have measured calcium and magnesium transport along the inner medullary collecting duct (IMCD) in rats by the microcatheterization technique. The fraction of filtered divalent cation ((F/P)Ca,Mg/In) as a function of IMCD length was analyzed by linear regression. In eight control rats calcium reabsorption was demonstrated, inasmuch as there was a significant correlation and slope, P less than 0.001. (F/P)Ca/In decreased from 2.2 +/- 0.3% at the beginning of the IMCD to 0,84 +/- 0.1% in the urine. (F/P)Mg/In was not changed along the IMCD, After acute thyroparathyroidectomy (TPTX) (F/P)Ca/In increased to 4.6 +/- 0.4% at the beginning of the IMCD and 1.55 +/- 0.1% in the urine. The slope was not significantly different from the control group. (F/P)Mg/In was unchanged and was not different from the control group. Accordingly, net Ca reabsorption occurs along the IMCD and the fractional Ca reabsorption is not altered by TPTX. Net Mg transport along the IMCD was absent in both control and TPTX rats; The increased calciuresis after TPTX is not dependent on IMCD function nor is Mg excretion under the conditions studied.
PTH inhibition of bicarbonate transport by proximal convoluted tubules
Authors: McKinney T. D. , Myers P.
Published: December 2017

These studies examined the effect of parathyroid hormone (PTH), dibutyryl cyclic AMP DBcAMP, and 8-bromo-cyclic AMP BrcAMP on HCO3- transport by rabbit superficial proximal convoluted tubules perfused in vitro. Bicarbonate was estimated as total CO2 measured microcalorimetrically. At slow perfusion rates with 25 mM HCO3- in the perfusate and bath, PTH (0.1 U/ml in the bath) caused the total CO2 in tubular fluid to rise from 10.2 to 19.9 mM. The hormone had no effect on the total CO2 concentration in tubules perfused with HCO3(-)-free perfusates. With HCO3(-) in the perfusate and bath, PTH reduced the rates of fluid and total CO2 absorption to 57 and 48% of control values, respectively. PTH had no effect on the rates of fluid absorption and total CO2 secretion when HCO3(-)-free perfusates were used. The effects of DBcAMP and BrcAMP (10(-7) M in the bath) were similar to those of PTH. 5'-AMP (10(-6) M in the bath) did not alter the total CO2 concentration of tubular fluid when the tubules were perfused at slow rates with HCO3- in the perfusate and bath. Ouabain (10(-5) M in the bath) caused the total CO2 concentration in tubules perfused at slow rates with HCO3--free perfusates to rise from 8.9 to 12.7 mM. PTH caused no further change in the total CO2 concentration in the presence of ouabain.
HCO3-Cl exchange transport in the adaptive response to alkalosis by turtle bladder
Authors: Cohen L.
Published: December 2017

The isolated turtle urinary bladder acidifies its mucosal (M) solution, and the rate of acidification (JH) is equivalent to the short-circuit current after Na+ transport is abolished by ouabain. When HCO3(-) is present in the serosal solution it is secreted into M in an electroneutral exchange for absorbed Cl-. The rate of HCO3(-) secretion (JHCO3(-)) can be measured by pH stat titration after JH is nullified by an opposing pH gradient. With use of these methods JH and JHCO3 were measured sequentially in bladdes from control animals and animals fed NaHCO3 (alkalosis) or NH4Cl (acidosis). JH in alkalosis (57 +/- 6 micro A) was ot different from control values (53 +/- 7 micro A). JHCO3, however, was nearly 40% higher in alkalosis (1.63 +/- 0.11 vs. 1.17 +/- 0.14 mu mol x h-1 x 8 cm-2). In contrast, JHCO3 in acidosis was similar to control values (0.89 +/- 0.15 mu mol x h-1 x 8 cm-2) but JH was increased. As judged from Cl- fluxes, neither alkalosis nor acidosis altered the electroneutral coupling between HCO3(-) secretion and Cl- absorption. JH and JHCO3 appear to be independent processes in the turtle bladder that are capable of responding independently to physiologic changes in the acid-base status of the intact animal.
Glomerular ultrafiltration dynamics: euvolemic and plasma volume-expanded rats
Authors: Arendshorst W. J. , Gottschalk C. W.
Published: December 2017

With the use of micropuncture techniques, glomerular dynamics were characterized in euvolemic Munich-Wistar (Chapel Hil) rats. Single nephron glomerular filtration rate (SNGFR) and plasma flow (SNGPF) averaged 31 nl/min and 100 nl/min, respectively. Efferent effective ultrafiltration pressure (PUF) was greater than zero in each animal (7 mmHg, P less than 0.001), indicating filtration pressure disequilibrium. Mean PUF (PUF) was 15 mmHg and specific glomerular filtration coefficient (Kf) values averaged 0.033 nl/(s x mmHg). Similar experiments were performed on Munich-Wistar rats from B.M. Brenner's colony. Filtration pressure equilibrium was reached in rats with SNGPF < 112 nl/min; disequilibrium existed when SNGPF was > 168 nl/min. Overall, PUF (15 mmHg) was similar to that in Chapel Hill rats, SNGFR (49 nl/min) was greater, as was specific Kf (0.066 nl/(s.mmHg)). These observations document important Kf differences in the two colonies. In other experiments on Chapel Hill rats, plasma volume expansion produced increases in SNGPF and renal plasma flow (RPF). However, SNGFR (33 nl/min) and GFR (1.2 ml/(min x g kidney wt)) were unchanged, as were SNGFR/GFR and SNGPF/RPF ratios. SNGFR was relatively insensitive to increased plasma flow because of the relatively low Kf and filtration pressure disequilibrium during euvolemia. In additional experiments, GFR was also only weakly dependent on RPF in conscious rats.
Effects of phosphate loading and parathyroid hormone on starling renal phosphate excretion
Authors: Wideman R. F. , Clark N. B. , Braun E. J.
Published: December 2017

The effects of phosphate loading were examined in intact, parathyroidectomized (PTX), and PTX starlings infused with parathyroid extract (PTX +/- PTE). In intact starlings, there was no correlation between the filtered phosphate load and the quantity of phosphate excreted. Net secretion predominated at lower filtered loads (2.5-20 mu mol . kg-1 . min-1) but net reabsorption occurred at filtered loads between 20 and 35 mu mol . kg-1 . min-1. Phosphate-loaded PTX starlings reabsorb phosphate over a wide range of filtered loads (5-45 mu mol . kg-1 . min-1). Excretion was significantly correlated with the filtered load in PTX birds (r = 0.943). Phosphate loaded PTX + PTE starlings secreted phosphate when filtered loads ranged from 2.5 to 30 mu mol . kg-1 . min-1. Excretion also was significantly correlated with the filtered load in this group (r = 0.972). The results of these experiments indicate that a) variations in endogenous PTH release in intact starlings result in wide variations in the renal pattern of phosphate excretion during phosphate loading; b) net phosphate secretion rarely occurs during phosphate loading in PTX starlings, but always occurs in PTX + PTE starlings; and c) both phosphate secretion and phosphate reabsorption are limited by maximal tubular transport capacities. Additional evidence indicates that PTH inhibits tubular phosphate reabsorption in starlings.
Interaction between antidiuretic and parathyroid hormones on urine concentration
Authors: Humes H. D. , Simmons C. F. , Brenner B. M.
Published: December 2017

Experiments were performed on 26 acutely thyroparathyroidectomized (TPTX) Sprague-Dawley rats undergoing maximum water diuresis to determine whether the rise in urinary osmolality (Uosmol) in response to a submaximal dose of antidiuretic hormone (ADH) is modified by exogenous administration of parathyroid hormone (PTH). During administration of a submaximal dose of PTH to 11 TPTX rats, the ADH-induced increase in Uosmol averaged 267 +/- 15 mosmol, or twice the average increment of 131 +/- 18 mosmol observed when the same dose of ADH was given prior to PTH infusion (P < 0.001). This difference could not be attributed to changes in endogenous ADH release, renal hemodynamics, or solute excretion, and was not observed in a second group of eight other water-diuretic TPTX rats given sham PTH infusion. A third group of seven water-diuretic TPTX rats were studied with verapamil, a compound known to antagonize calcium ion entry into cells. Pretreatment of these rats with intravenous verapamil abolished the PTH potentiation of the Uosmol response to ADH described above. We conclude, therefore, that PTH enhances the Uosmol response to ADH, perhaps via a mechanism requiring a PTH-mediated change in the cellular calcium concentration or content of cells important in the urinary concentrating process.
Electrical properties of the medullary collecting duct in the rat
Authors: Hayslett J. P. , Backman K. A. , Schon D. A.
Published: December 2017

Studies were performed in the rat to determine the transepithelial potential difference (PD) of the medullary collecting duct (MCD). With the microcatheter as an exploring electrode the PD from duct lumen to the reference electrode, placed in peritoneal fluid, was approximatley 1 mV, lumen positive, in the MCD. It was unlikely that the polarity and small magnitude of PD was due to an artifact created by the microcatheter, since essentially similar results were obtained by transmural puncture of the inner third of the duct using Ling-Gerard glass electrodes and glass microelectrodes, tip OD 3-5 micron. Moreover, a PD of a few millivolts, lumen positive, was observed in experimental states characterized by marked increases in potassium and hydrogen ion secretion and sodium absorption. Further studies showed that the diffusion potential existing within the interstitial fluid compartment, between papilla and reference electrode, was approximately +2 mV, owing to the higher NaCl concentration in the papillary area. These data demonstrate, therefore, that the transepithelial PD in MCD approaches zero in control animals, and is not substantially altered by marked changes in net electrolyte transport.
CO2 equilibria in renal tissue
Authors: Malnic Gerhard
Published: December 2017

Disequilibrium situations within the bicarbonate/CO2 buffer system have been observed in the kidney. On the one hand, a Pco2 difference between final urine and arterial blood has been detected. On the other, a disequilibrium pH was found in cortical renal tubules and attributed to either increased carbonic acid concentrations or to Pco2 levels above those of arterial blood. Recent methodological developments have yielded better insight into these disequilibrium situations. They include, besides the use of pH microelectrodes, the introduction of microcalorimetric total CO2 determinations and Pco2 microelectrodes. There is agreement concerning the finding of a Pco2 10–40 mmHg above that in arterial blood in renal cortical tubules; however, stellate vessel Pco2 equal to tubular Pco2 was found by only one group of investigators. According to their view, diffusion equilibrium exists between all cortical structures, a finding disputed by others using a similar methodology. CO2 permeability of tubular epithelium is also still controversial, with values ranging from one-half (microcalorimetric method) to one-twentieth (pH-equilibration method) of an equivalent water layer having been reported. Two positions emerge on the basis of recent experimental results: the existence of very high permeability to CO2 and consequent diffusion equilibrium in renal cortex, and the occurrence of CO2 diffusion limitation leading to the establishment of finite Pco2 gradients across renal tubular epithelium.

disequilibrium pH; CO2 permeability; tubular Pco2; CO22 fluxes; carbonic anhydrase
Chloride concentration gradient in newborn dogs in the presence of distal nephron blockade
Authors: Haramati Aviad , Kleinman Leonard I.
Published: December 2017

Renal tubular Na+, Cl-, and H2O reabsorption was determined in 14 newborn dogs, 3–29 days of age, and in three adult dogs. In all animals NaCl reabsorption beyond the proximal tubule was blocked with ethacrynic acid (2 mg/kg) and amiloride (2.4 mg/kg). During distal blockade, fractional reabsorption of NaCl and water in both newborns and adults was approximately 70%, and there was a urine-to-plasma chloride gradient equal to 1.34 ± 0.01, indicating that the proximal tubules of the newborn as well as those of the adult can generate a transtubular Cl- gradient. Upon administration of acetazolamide (50 mg/kg), there was a dramatic increase in excretion of Na+, Cl-, HCO3-, and water, and a decrease in the transtubular chloride gradient. After acetazolamide, the degree of inhibition of HCO3- reabsorption was well correlated with that of Na+ (r = 0.77) or Cl- (r = 0.74), and Na+ or Cl- inhibition exceeded that of HCO3- In the newborn animal, the ratio of inner-to-outer cortical nephron function is high at birth and declines rapidly during the first few weeks of life. However, there was no correlation between age and changes in either electrolyte excretion or in the transtubular chloride gradient. Therefore, the newborn dog possesses Cl- permselective tubules in the inner cortex that, in the presence of intact HCO3- reabsorption, are capable of establishing a functional transtubular Cl- gradient contributing to NaCl reabsorption.

bicarbonate reabsorption; sodium reabsorption; ethacrynic acid; amiloride; acetazolamide; proximal tubule

Submitted on October 29, 1979

Accepted on April 18, 1980
Juxtamedullary and superficial nephron phosphate reabsorption in the cat
Authors: Goldfarb Stanley
Published: December 2017

The cat kidney possesses discrete venous drainage systems for superficial and deep (juxtamedullary) portions of the renal cortex. Arteriovenous (A-V) extraction and micropuncture studies may thus be used to evaluate PO4 transport in the two nephron populations. Determinations of A-V extraction showed whole kidney percentage of fractional PO4 excretion (%FEPOPO4) was 29.05 ± 2.01%, percentage of fractional delivery of PO4 (%FDPOPO4) from the superficial nephrons was 42.94 ± 2.42%, and that from deep nephrons was 22.35 ± 2.05%. The %FDPOPO4 from superficial nephrons was significantly greater than %FEPOPO4 in urine (P < 0.001) and than %FDPOPO4 from deep nephrons (P < 0.001). Micropuncture studies showed %FDPOPO4 from late distal tubules was 41.63 ± 3.61%, while %FEPOPO4 in urine was 30.24 ± 2.22%. Infusion of acetazolamide increased both %FEPOPO4 in urine and %FDPOPO4 from deep nephrons (A-V extraction studies), but had no effect on %FDPOPO4 from either superficial nephrons (A-V extraction) or from late distal tubules (micropuncture), thus abolishing the differences between superficial and deep compartments. Nephron heterogeneity apparently exists for superficial and juxtamedullary PO4 reabsorption in the cat. Acetazolamide increased %FEPOPO4 in urine primarily by reducing deep nephron PO4 reabsorption.

nephron heterogeneity; micropuncture; acetazolamide

Submitted on November 13, 1979

Accepted on April 10, 1980
Effect of dietary phosphate on transport properties of pig renal microvillus vesicles
Authors: Barrett Paula Q. , Gertner Joseph M. , Rasmussen Howard
Published: December 2017

Dietary phosphate manipulation results in stable adaptive changes in the transport functions of microvillus membrane vesicles isolated from pig renal cortex. When assayed under sodium gradient conditions, phosphate uptake is enhanced 200–400% in vesicles prepared from animals maintained on a low-phosphate diet (0.22%) compared to high-phosphate diet controls (0.82%). When transport is assayed in sodium preequilibrated vesicles, a 100% enhancement of phosphate uptake is demonstrable. Stimulation of phosphate uptake into low-phosphate diet vesicles after the imposition of a sodium chloride gradient is equivalent if uptake is measured at pH 6.0 or 8.0 and can be kinetically characterized as resulting from a Vmax alteration in the phosphate transport system. Microvillus membrane vesicle phosphate transport is maximally stimulated after only 2 days of dietary deprivation. Although a longer period (1 and 2 wk) of phosphate restriction does not further stimulate phosphate transport, it does result in an inhibition of other sodium gradient-dependent transport systems (glucose, alanine).

phosphate transport; phosphate restriction; glucose transport; brush-border vesicles

Submitted on October 22, 1979

Accepted on April 25, 1980
Cell pH and acid transport in renal cortical tissue
Authors: Kleinman J. G. , Brown W. W. , Ware R. A. , Schwartz J. H.
Published: December 2017

Cell pH (pHc) was examined by the [14C]DMO technique in suspensions of proximal tubule fragments from rabbit renal cortex. In buffer with 10 mM HCO3(-), pHc was more alkaline than external pH (pHe) at values of the latter < 7.4. Maximal cell-to-extracellular pH gradients (delta pH) occurred at pHe = 6.8 and below. At pHe > 7.4, pHc was more acid than pHe was. However, pHc was always more alkaline than the electrochemical equilibrium pH. At pHe congruent to 7.0, 60 min of deoxygenation decreased delta pH from 0.22 +/- 0.02 to 0.05 +/- 0.01. Reoxygenation restored delta pH to control values. Incubation with ouabain abolished the delta pH. Both the carbonic anhydrase inhibitor, acetazolamide, and the anion transport inhibitor, 4-acetamido-4'-isothiocyano-2,2'-disulfonic stilbene (SITS), increased delta pH. The studies demonstrate relative intracellular alkalinity in proximal tubule. A fall in pHc occurs with maneuvers that interfere with H+ pumping out of the cells. A rise in pHc occurs with maneuvers that interfere with the disposition of intracellular alkali: slowing of HCO3(-) generation with acetazolamide or blocking of HCO3(-) exit with SITS. The results support a H+-secretory model of proximal tubule acid transport that is dependent on maintenance and dispersal of intracellular alkalinity.