Biomedical subjects

J B Stokes

Publications and source records attributed to J B Stokes.

At least 73 records · Page 4Linked to original sources

PubMed1989

Reduction in sensitivity to Cl- channel blockers by HCO3- -CO2 in rabbit cortical collecting duct.

We examined the ability of HCO3- -CO2 to modify the potency of Cl- channel blockers in the renal cortical collecting duct (CCD) for the following two reasons. 1) From a practical point of view, there is, to our knowledge, no information regarding the effect of the HCO3- -CO2 buffer system on the potency of Cl- channel blockers. 2) We showed in the companion manuscript [Am. J. Physiol. 257 (Cell Physiol. 26): C94-C101, 1989] that HCO3- -CO2 stimulates transepithelial anion exchange in the CCD. Based on precedent in the literature, we postulated that HCO3- stimulates the basolateral membrane Cl- conductance. Here, we demonstrate that several Cl- channel blockers can reduce CCD Cl- self exchange when the solutions are buffered in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES). Concentrations of blockers producing 80% inhibition in HEPES, pH 7.4, produced only 20% inhibition in 25 mM HCO3- -CO2, pH 7.4. The ability of HCO3- -CO2 to reduce blocker potency had an IC50 of only 2 mM. We also examined interactions of HCO3- -CO2 and blockers with regard to the principal cell basolateral Cl- conductance. Blockers did not alter the Rb+ flux, a marker of K+ transport, but did reduce transepithelial conductance (GT), i.e., the blockers inhibited the principal cell basolateral Cl- conductance. As was the case with intercalated cell anion exchange, GT measurements indicated that HCO3- -CO2 impaired the ability of Cl- channel blockers to inhibit the principal cell Cl- conductance.(ABSTRACT TRUNCATED AT 250 WORDS)

Amiloride↗

PubMed1989

Stimulation of Cl- self exchange by intracellular HCO3- in rabbit cortical collecting duct.

In rabbit cortical collecting duct, Cl- self exchange accounts for most of the transepithelial Cl- tracer rate coefficient, KCl (nm/s); a small fraction is effected by Cl--HCO3- exchange and Cl- diffusion. We previously reported that changing from a CO2-free N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) bath to a 5% CO2-25 mM HCO3- bath stimulates Cl- self exchange. Here, we examine in further detail the individual components of the CO2-HCO3- system that stimulate KCl. Addition of 0.5% CO2 to a HEPES bath (final pH = 7.24) stimulated KCl by 70 +/- 19 nm/s, a delta KCl comparable to that induced by 1% CO2 (pH 7.12), 6% CO2 (pH 6.6), or 6% CO2-25 mM HCO3- (pH 7.4). The roles of intracellular pH (pHi) and HCO3- concentration were examined by clamping pHi using high K+ and nigericin. Increasing pHi from 6.9 to 7.6 in solutions without exogenous CO2 or HCO3- increased KCl by 71 +/- 17 nm/s. These results suggest that pHi might regulate anion exchange. However, during such a pHi-shift experiment, metabolically derived CO2 produces a concomitant change in intracellular HCO3- concentration [( HCO3-]i). To determine whether an increase in [HCO3-]i could stimulate Cl- self exchange, we replaced HEPES with 6% CO2-5 mM HCO3- isohydrically (pHi clamped at 6.9). With this increase in [HCO3-]i at constant pHi, KCl increased by 51 +/- 10 nm/s. These maneuvers had negligible effects on Cl- diffusion and Cl--HCO3- exchange. These experiments demonstrate that increases in cell [HCO3-] (or perhaps CO2) can stimulate transepithelial anion exchange.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1989

K+ and Rb+ transport by the rabbit CCD: Rb+ reduces K+ conductance and Na+ transport.

We compared transport of K+ and Rb+ across the rabbit cortical collecting duct to gain insight into the mechanisms of K+ secretion. Passive tracer fluxes, active secretory rates, electrophysiological behavior, and the ability of each ion to support Na+-K+-ATPase activity were determined. When active transport was inhibited by amiloride, K+ permeability was twice the Rb+ permeability. Transepithelial conductance (GT) was half as great in solutions where 5 mM Rb+ replaced 5 mM K+. When 4 mM Ba2+ was added to the lumen, both Rb+ and K+ permeability fell to values not different from that expected for paracellular diffusion. The relationship between Ba2+-induced changes in the K+ and Rb+ permeabilities and in the simultaneously measured GT provides strong evidence that K+ transport across the apical membrane is largely, if not exclusively, conductive. We also determined that net K+ secretion is greater than net Rb+ secretion (when each is the abundant ion). The reasons for this difference probably involve several steps in the K+ secretory process and include the following: 1) reduced ATPase activity in the presence of Rb+ (approximately 80%) compared with K+, 2) reduction of Na+ absorption, and 3) partial blockade of the apical (and perhaps basolateral) K+ conductance. Although there were quantitative differences between K+ and Rb+ transport, we found no evidence suggesting that these ions are transported by different mechanisms.

Absorption↗

PubMed1988

Absence of transepithelial anion exchange by rabbit OMCD: evidence against reversal of cell polarity.

In the rabbit cortical collecting duct (CCD), Cl tracer crosses the epithelium predominantly via an anion exchange system that operates in either a Cl-Cl or Cl-HCO3 exchange mode. In the present study, we used the 36Cl lumen-to-bath rate coefficient (KCl, nm/s), a sensitive measurement of CCD transepithelial anion transport, to investigate the nature of Cl transport in the medullary collecting duct dissected from inner stripe, outer medulla (OMCD). The KCl in OMCD perfused and bathed in HCO3-Ringer solution was low (46.2 +/- 8.5 nm/s) and similar to that value observed in the CCD when anion exchange is inhibited and Cl permeates the epithelium by diffusion. Unlike KCl in CCD, KCl in OMCD was not stimulated by adenosine 3',5'-cyclic monophosphate (cAMP). OMCD KCl was not altered by bath Cl and/or HCO3 removal, demonstrating the absence of transepithelial Cl-Cl and Cl-HCO3 exchange. To test the hypothesis that metabolic alkalosis could reverse the polarity of intercalated cells and thus induce an apical Cl-HCO3 exchanger in H+-secreting OMCD cells, we measured KCl in OMCD from rabbits made alkalotic by deoxycorticosterone and furosemide. Although the base-line KCl was slightly higher than in OMCD from control rabbits, the value was still far lower than the KCl under comparable conditions in CCD. Moreover, KCl in OMCD from alkalotic rabbits was unchanged by cAMP, or by sequential removal of bath HCO3 and Cl. Immunocytochemistry using peanut lectin and a monoclonal antibody to-erythrocyte band 3 failed to reveal any evidence for alkalosis-induced reversal of either CCD or OMCD intercalated cell polarity.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkalosis↗

PubMed1988

Passive NaCl transport in the flounder urinary bladder: predominance of a cellular pathway.

The urinary bladder of the winter flounder is a high-resistance epithelium that can absorb Na and Cl in an electrically silent manner. This active absorption (mucosa-to-serosa) of NaCl is, apparently uniquely, inhibited by mucosal hydrochlorothiazide (HCTZ). These experiments evaluated the notion that virtually all of the cellular Na and Cl permeation could be inhibited by mucosal HCTZ. Mucosal Ba2+ reduced the transepithelial conductance from 0.74 +/- 0.08 to 0.60 +/- 0.06 mS/cm2. Mucosal HCTZ reduced the serosa-to-mucosa flux (backflux) of Na from 0.70 +/- 0.08 to 0.29 +/- 0.03 mueq.cm-2.h-1 and the backflux of Cl from 1.92 +/- 0.22 to 0.38 +/- 0.03 mueq.cm-2.h-1. The treatment with these two agents caused the sum of the partial ionic conductances for Na and Cl to approximate the measured transepithelial conductance. In response to the imposition of a transepithelial voltage, the HCTZ-insensitive Na and Cl backfluxes behaved largely as predicted by the laws of simple ionic diffusion, although there was still a detectable cellular backflux. As judged from dilution voltages and tracer fluxes, the diffusional (paracellular) pathway(s) is nonselective for Na and Cl. The HCTZ-sensitive cellular Na and Cl backfluxes are dependent on the presence of mucosal Na and Cl. Neither backflux is significantly inhibited by serosal application of commonly used inhibitors of Na or Cl transport. The results demonstrate that the majority of passive Na and Cl flux is via a cellular pathway. The translocation across the apical membrane probably involves the same NaCl cotransport process responsible for NaCl absorption.

Animals↗

PubMed1988

Characteristics of the paracellular pathway of rabbit cortical collecting duct.

The nature of the paracellular pathway of the rabbit cortical collecting duct (CCD) was examined under conditions designed to eliminate all cellular ion transport. Transcellular conductive pathways were blocked by addition of amiloride and Ba2+ to the perfusate. Cl self-exchange was eliminated by removing Cl from the bath solution, and HCO3 transport was eliminated by omitting HCO3 and CO2 from the solutions. The residual transepithelial conductance (GT) and radioisotopic tracer flux under these conditions probably occur via the paracellular pathway. The GT measured in nontransporting CCD bathed in NaCl solutions was 1.1-1.2 mS/cm2. When Na or Cl was replaced by a less mobile ion, the GT decreased by an amount commensurate with the decrease in solution conductivity. The Na-to-Cl permeability ratio determined by NaCl dilution voltages ranged from 0.55 to 0.82. An independent estimate of paracellular selectivity was obtained by comparing the lumen-to-bath tracer rate coefficients for Na (kNa) and Cl (kCl). The ratio kNa:kCl was 0.75. These observations suggest that the paracellular pathway displays a Na:Cl permselectivity not substantially different from the ratio of their mobilities in water (0.65). Additional experiments demonstrated that the summed partial ionic conductances of Na and Cl calculated from the tracer fluxes were in close agreement with the measured GT. We conclude that the paracellular conductance of CCD is nonselective in character and approximately 1-2 mS/cm2 in magnitude.

Animals↗

PubMed1988

Characteristics of papillary collecting duct cells in primary culture.

We examined the electrophysiological and Na+ transport characteristics of rat papillary collecting duct (PCD) cells grown in primary cultures. Grown as monolayers on polycarbonate filters, the cells displayed similar morphological characteristics to native epithelia. They also bound Dolichus biflorus lectin, a property shared by native cells. Monolayers developed a peak electrical resistance of 100-200 omega.cm2 and a transmonolayer voltage of less than 2 mV. Similar values were measured in the perfused, native PCD of the same species as well as PCD cells cultured from rabbit and bovine kidneys. Hamster cells did not readily develop confluent monolayers under the same conditions. Exposure of the cultured cells to 10% fetal calf serum for 24 h caused the Na+ uptake across the apical membrane to double, an effect not reproduced by indomethacin, insulin, vasopressin, aldosterone, dexamethasone, or hexamethylene bisacetamide (an inducer of differentiation). Amiloride (1 mM) inhibited Na+ uptake by 50-80%. The measured short-circuit current did not correlate with Na+ uptake and was clearly dissociated by exposure to serum. The results suggest that there is more than one mechanism of ion transport by the rat PCD.

Amiloride↗

PubMed1988

Inhibition of cortical collecting tubule chloride transport by organic acids.

Cl self-exchange by the rabbit cortical collecting tubule (CCT) occurs via an apical anion exchanger in series with a basolateral Cl conductance. We studied the effects of organic acids on CCT Cl self-exchange. We found no evidence for transport of acid anions by the self-exchange system. Rather, Cl self-exchange was inhibited by a variety of organic acids. The degree of inhibition correlated with the chloroform/water partition coefficient and was enhanced by lowering pH, indicating inhibition by the lipid-soluble, protonated species. Inhibition by the representative acid iso-butyrate was dose-dependent and showed sidedness (basolateral greater than apical). Iso-butyrate also reversibly reduced transepithelial conductance without altering K permeability, suggesting inhibition of the principal cell basolateral Cl conductance. Because small organic compounds with similar lipid solubilities but no carboxyl group had no effect, both the carboxyl group and the lipid-solubility of organic acids appear to be important. The results are consistent with blockade of chloride channels by organic acids.

1-Butanol↗

PubMed1987

Chloride transport by the cortical and outer medullary collecting duct.

The processes by which chloride is transported by the cortical and outer medullary collecting tubule have been most extensively studied using in vitro microperfusion of rabbit tubules. Chloride appears to be transported by three major mechanisms. First, Cl can be actively reabsorbed by an electroneutral Cl-HCO3 exchanger localized to the apical membrane of the HCO3-secreting (beta-type) intercalated cell. Cl exits this cell via a basolateral Cl channel. This anion exchange process can also operate in a Cl self-exchange mode, is stimulated acutely by beta-adrenergic agonists and cAMP, and is regulated chronically by in vivo acid-base status. Second, Cl can diffuse passively down electrochemical gradients via the paracellular pathway. Although this pathway does not appear to be selectively permeable to Cl, it is large enough to allow for significant passive reabsorption. Third, Cl undergoes recycling across the basolateral membrane of the H+-secreting (alpha-type) intercalated cell. HCO3 exit from this cell brings Cl into the cell via electroneutral Cl-HCO3 exchange; Cl then exits the cell via a Cl channel. Cl transport is thus required for acidification and alkalinization of the urine. Both of these processes exist in the cortical collecting tubule. Their simultaneous operation allows fine tuning of acid-base excretion. In addition, these transport systems, when functioning at equal rates, effect apparent electrogenic net Cl absorption without changing net HCO3 transport. These systems may play an important role in regulating Cl balance.

Animals↗

PubMed1987

Purification of rat papillary collecting duct cells: functional and metabolic assessment.

Rat renal papillary collecting duct (PCD) cells were isolated using collagenase and hyaluronidase digestion and a three-step low-speed centrifugation. As assessed by binding of the lectin Dolichos biflorus and determination of vasopressin-sensitive adenylate cyclase and Na+-K+-ATPase, the enrichment of PCD cells over a crude papillary cell preparation was 1.8, 2.4, and 1.4, respectively. Microscopic evaluation indicated that the preparation was greater than 90% pure PCD cells. The isolated cells were viable as evident from the high K/Na ratio of intracellular electrolytes measured by electron probe analysis (5.3), from the high ATP/ADP ratio (2.15), and the metabolic response to alterations in Na transport. Exposure to 2 mM ouabain or removal of Na reduced O2 consumption by 25-35%; the uncoupler carboxylcyanide-m-chlorophenylhydrazone more than doubled O2 consumption. In the presence of 14 mM glucose and at a PO2 of 100 Torr the cells produced substantial quantities of lactate. This aerobic glycolysis may account for greater than 20% of the ATP production. In the presence of rotenone, glycolysis increased by 56% and was able to maintain the cellular ATP level at 65% of control. In the absence of any exogenous substrate PCD cells respired normally and had a close to normal ATP content, but lactate production was markedly decreased. These results demonstrate that viable PCD cells can be isolated from rat kidney. At normal PO2 and in the presence of D-glucose the cells show a substantial amount of aerobic glycolysis, although their mitochondrial respiration is not rate limiting. In the absence of glucose the cells derive the majority of their energy from an as yet unidentified endogenous substrate.

Adenosine Triphosphate↗

PubMed1986

Variability of functional characteristics of MDCK cells.

We measured several functional parameters of MDCK cells cultured as monolayers in order to more fully characterize their ion transport properties. Most of the present studies were completed with five groups (A-E) of MDCK cells studied from passage 62 to 78. Each group represents the same subline of MDCK cells after having been frozen, stored, and thawed at passage 62 or 64. The median transmonolayer resistances of the groups were 507, 149, 284, 72, and 126 omega X cm2. Addition of amphotericin B to the apical solution induced a oubain-sensitive transepithelial current. The apical membrane voltage and fractional resistance exhibited a wide range of values in two of the groups studied, with mean values of -32 mV and 0.68 in group B and -40 mV and 0.78 in group E. Neither apical nor basolateral membrane displayed significant Na+ conductance. K+ conductance was present in the basolateral but not in the apical membrane. Acidification or alkalinization of the apical solution was dependent on the conditions used to study the cells. The 4,4'-diisothyocyano-2,2'-disulfonic stilbene inhibited acidification (or induced alkalinization), whereas increasing ambient HCO-3 concentration induced alkalinization. The results of these studies indicate qualitatively similar behavior between five groups of MDCK cells but significant quantitative differences between the groups. Analysis of the variability of the measured parameters indicated that there were no differences as a function of passage number within a group. The factors responsible for functional differences between groups are not known but may be related to the cell storage process.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed1986

Patterns of K+ permeation following inhibition of Na+ transport in rabbit cortical collecting tubule.

The passive (lumen-to-bath) K+ permeation (KK) of rabbit cortical collecting tubules was measured before and after inhibition of Na+ transport. Inhibition of the Na-K pump with ouabain reduced KK. This result contrasts sharply with the previously described increase in KK observed following inhibition of Na+ transport with amiloride. These opposite changes in KK are owing to the fact that a substantial component of the lumen-to-bath K+ permeation involves a transcellular pathway. Amiloride, because it hyperpolarizes the apical membrane, increases KK; ouabain, because it depolarizes the cell, decreases KK. Previous results have also suggested that the cell K+ permeability is secondarily altered by these agents so that the changes in voltage and permeability are additive. These patterns of changes in KK were used to evaluate the mechanism of action of two agents that partially inhibit Na+ transport: vasopressin and prostaglandin (PG) E2. Their effect on KK was qualitatively similar to that of amiloride. In amiloride-treated tubules, neither vasopressin nor PGE2 altered KK. Neither did they alter the normal reduction in KK caused by pump inhibition. Thus they did not have any direct effect on K+ permeability. These results are consistent with the thesis that vasopressin and PGE2 inhibit Na+ absorption by reducing apical membrane permeability. The relation between the regulation of Na+ absorption and K+ permeation may have important implications for the regulation of K+ secretion by the cortical collecting tubule.

Amiloride↗

PubMed1986

Regulation of chloride self exchange by cAMP in cortical collecting tubule.

The hormonal control of Cl transport was examined in rabbit cortical collecting tubules using the lumen-to-bath 36Cl tracer rate coefficient (KCl, nm/s). Tracer movement via Cl-HCO3 exchange was minimized by using HCO3-CO2-free solutions. The electrical driving force was minimized by treating with amiloride. Under these conditions, net Cl transport was zero, yet there was a large KCl that fell 88% on removing bath (trans) Cl. These results are consistent with the mechanism of tracer flux being predominantly Cl self exchange. KCl fell spontaneously with time in vitro; after this decline KCl could be stimulated with 8-bromo-cAMP. cAMP present from the onset of perfusion prevented the time-dependent fall in KCl. When tracer movement was restricted to diffusion by eliminating Cl self exchange (0 Cl bath), cAMP had no effect on KCl. Although both isoproterenol and vasopressin are known to stimulate adenylate cyclase in this epithelium, only isoproterenol mimicked the cAMP effect on KCl. The isoproterenol effect was blocked by either propranolol or prostaglandin E2. Lumen addition of the disulfonic stilbene DIDS had no effect on KCl. Lumen addition of furosemide or trichloromethiazide had minimal or no effect. Taken together, these results indicate that Cl self exchange is regulated by beta-adrenergic agents acting via cAMP. The lack of an effect of vasopressin suggests cellular heterogeneity in this response to cAMP.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed1986

Stimulation of chloride transport by HCO3-CO2 in rabbit cortical collecting tubule.

We examined both the role of HCO3-CO2 in Cl transport as well as the effect of in vivo acid-base status on Cl transport by the rabbit cortical collecting tubule. The lumen-to-bath 36Cl tracer flux, expressed as the rate coefficient KCl, was measured in either HEPES-buffered (CO2-free) or HCO3-CO2-containing solutions. Amiloride was added to the perfusate to minimize the transepithelial voltage and thus the electrical driving force for Cl diffusion. Because KCl fell spontaneously with time in HCO3-CO2 solutions in the absence but not the presence of cAMP, we used cAMP throughout to avoid time-dependent changes. Acute in vitro removal of bath HCO3-CO2 reduced KCl. Acetazolamide addition in HEPES-buffered solutions also lowered KCl; KCl could be restored to control values by adding exogenous HCO3-CO2 in the presence of acetazolamide. In vivo acid-base effects on Cl transport were determined by dissecting tubules from either NaHCO3-loaded or NH4Cl-loaded rabbits. Tubules from HCO3-loaded rabbits had higher rates of Cl self exchange. Acute in vitro addition of bath HCO3-CO2 increased KCl and did so to a greater degree in tubules from HCO3-loaded rabbits. Most of this effect of HCO3-CO2 addition on KCl could not be accounted for by Cl-HCO3 exchange; rather, it appeared due to stimulation of Cl self exchange. The data are consistent with 36Cl transport occurring via Cl-HCO3 exchange as well as Cl self exchange. Both processes are acutely stimulated by HCO3 and/or Co2, and both are chronically regulated by in vivo acid-base status.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1986

Effects of inhibitors of Cl conductance on Cl self-exchange in rabbit cortical collecting tubule.

Electroneutral vs. conductive pathways of Cl transport were examined by measuring transepithelial conductance (GT) and the lumen-to-bath 36Cl rate coefficient (KCl). Experimental conditions minimized both Cl-HCO3 exchange [HCO3/CO2-free, N-2-hydroxyethylpiperazine-N'-2-ethane-sulfonic acid (HEPES)-buffered solutions] and the electrical driving force for paracellular Cl diffusion (amiloride in the perfusate, transepithelial voltage near zero). Two agents known to inhibit Cl conductances in other epithelia, anthracene-9-carboxylate (9AC, 1 mM) and diphenylamine carboxylate (DPC, 0.1-0.5 mM) reversibly reduced GT and KCl when added to the bath. Both reduced KCl to values consistent with paracellular diffusion. Bath DPC had no effect on GT in the presence of 4 mM lumen Ba2+, suggesting that the DPC-sensitive conductance is in series with an apical K conductance, i.e., resides on the basolateral membrane. Lumen DPC also reduced GT and KCl, but was less potent than bath DPC. Because the lumen DPC effect on GT was also blocked by lumen Ba2+, lumen DPC probably inhibits a basolateral Cl conductance. K removal and ouabain (0.5 mM) had no effect on KCl, suggesting that Cl tracer movement is not predominantly through the principal cell. We assume that these agents are inhibiting Cl conductive pathways and propose a model in which transcellular Cl movement through the intercalated cell occurs via an apical electroneutral entry step in series with a basolateral conductive pathway.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1985

Mineralocorticoid effect on K+ permeability of the rabbit cortical collecting tubule.

Mineralocorticoid hormones stimulate Na+ absorption and K+ secretion by the cortical collecting tubule. There is good evidence that this stimulation involves increasing luminal membrane Na+ permeability and the turnover rate (or number) of the Na+-K+ pumps. These experiments were designed to examine whether mineralocorticoid hormones also increase cell K+ permeability. Using 42K tracer measurements in tubules treated with amiloride to inhibit active Na+ and K+ transport, passive K+ permeation increased with increasing mineralocorticoid effect. Net Na+ absorption and the (passive) K+ efflux rate coefficient (KK) showed a linear relationship. The stimulatory effect was evident in vitro since 0.2 microM aldosterone added to the bath of tubules harvested from NaCl-loaded rabbits increased KK at 3 hrs while time controls showed no change. Since these tubules were also treated with amiloride, this increase in KK was not dependent on increasing Na+ absorption. The results indicate that in addition to the well-described effects of aldosterone on Na+ permeability and cell metabolism, the mineralcorticoid effect includes an increase in cellular K+ permeability.

Aldosterone↗

PubMed1985

Modulation of vasopressin-induced water permeability of the cortical collecting tubule by endogenous and exogenous prostaglandins.

These experiments were designed to test the thesis that prostaglandins produced by the cortical collecting tubule cells could modulate the vasopressin-induced osmotic water permeability (Pf). The dose-response curve for vasopressin-sensitive Pf showed the Km to be 1 microU ml-1. Exogenous PGE2 and PGF2 alpha (0.1 microM) inhibited the Pf induced by 1 microU ml-1 vasopressin when they were present in the bath solution. PGE2 (0.1 microM) in the lumen failed to inhibit the normal vasopressin-induced Pf, thus indicating an asymmetrical effect. Exposure of the tubule to 10 microM meclofenamate following stimulation of Pf by 0.2, 1.0, 10, or 100 microU ml-1 vasopressin failed to further increase the Pf. Pretreatment with meclofenamate or arachidonic acid (AA) failed to produce a different Pf response from controls. Neither naproxen (10 microM) nor AA altered significantly the Pf induced by 1 microU ml-1 vasopressin while methylisobutylxanthine, as expected, significantly enhanced Pf. The stable endoperoxide analogs U-44069 and U-46619, which mimic the actions of thromboxane A2 in many systems and which can stimulate osmotic water flow in the toad bladder, had no effect on Pf. Acidifying the lumen to pH 5.2 enhanced the Pf induced by 1 microU ml-1 vasopressin but subsequent exposure to meclofenamate did not cause an additional increment. These experiments demonstrate that exogenous prostaglandins are effective only from the basolateral surface of the cortical collecting tubule; that endogenous prostaglandins, if produced by these epithelial cells, do not produce demonstrable effects on vasopressin-sensitive Pf; and that endogenously produced thromboxane is not the likely reason for these results. Finally, the cortical collecting tubule response to many factors modulating Pf is considerably different from salientian urinary bladders.

1-Methyl-3-isobutylxanthine↗

PubMed1984

Pathways of K+ permeation across the rabbit cortical collecting tubule: effect of amiloride.

These experiments were designed to examine passive K+ transport by the rabbit cortical collecting tubule. Potassium diffusion voltages were used to assess the presence of apical and basolateral K+ conductances. With amiloride (0.1 mM) in the lumen, reproducible K+ diffusion voltages from both the lumen and bath were obtained. Amiloride enhanced the magnitude of these voltage deflections (delta VT). There were time-dependent increases in the K+ diffusion voltage but the steady-state values were highly reproducible in the same tubule. In the amiloride-treated tubules, delta VT induced by raising bath [K+] to 20 mM was larger than that produced by the same increase in lumen [K+]. To evaluate whether the amiloride-treated tubule had, as suggested by the K+ diffusion voltages, substantial K+ permeabilities on both apical and basolateral membranes, the K+ rate coefficient (lumen-to-bath, KK) was measured before and after amiloride treatment. The amiloride-induced increase in KK, from 66 +/- 6 to 205 +/- 35 nm/s, was significantly larger than could be accounted for by the changes in transepithelial voltage or membrane voltages alone. This discrepancy could be due to single-file diffusion across the apical membrane and/or the (secondary) enhancement of K+ permeability following inhibition of Na+ transport.

Amiloride↗