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D R Halm

Publications and source records attributed to D R Halm.

28 records · Page 2Linked to original sources
PubMed1985

Electrophysiology of flounder intestinal mucosa. I. Conductance properties of the cellular and paracellular pathways.

We evaluated the conductances for ion flow across the cellular and paracellular pathways of flounder intestine using microelectrode techniques and ion-replacement studies. Apical membrane conductance properties are dominated by the presence of Ba-sensitive K channels. An elevated mucosal solution K concentration, [K]m, depolarized the apical membrane potential (psi a) and, at [K]m less than 40 mM, the K dependence of psi a was abolished by 1-2 mM mucosal Ba. The basolateral membrane displayed Cl conductance behavior, as evidenced by depolarization of the basolateral membrane potential (psi b) with reduced serosal Cl concentrations, [Cl]s. psi b was unaffected by changes in [K]s or [Na]s. From the effect of mucosal Ba on transepithelial K selectivity, we estimated that paracellular conductance (Gp) normally accounts for 96% of transepithelial conductance (Gt). The high Gp attenuates the contribution of the cellular pathway to psi t while permitting the apical K and basolateral Cl conductances to influence the electrical potential differences across both membranes. Thus, psi a and psi b (approximately 60 mV, inside negative) lie between the equilibrium potentials for K (76 mV) and Cl (40 mV), thereby establishing driving forces for K secretion across the apical membrane and Cl absorption across the basolateral membrane. Equivalent circuit analysis suggests that apical conductance (Ga approximately equal to 5 mS/cm2) is sufficient to account for the observed rate of K secretion, but that basolateral conductance (Gb approximately equal to 1.5 mS/cm2) would account for only 50% of net Cl absorption. This, together with our failure to detect a basolateral K conductance, suggests that Cl absorption across this barrier involves KCl co-transport.

Animals↗
PubMed1985

Electrophysiology of flounder intestinal mucosa. II. Relation of the electrical potential profile to coupled NaCl absorption.

We characterized the hyperpolarization of the electrical potential profile of flounder intestinal cells that accompanies inhibition of NaCl cotransport. Several observations indicate that hyperpolarization of psi a and psi b (delta psi a,b) results from inhibition of NaCl entry across the apical membrane: (a) the response was elicited by replacement of mucosal solution Cl or Na by nontransported ions, and (b) mucosal bumetanide or serosal cGMP, inhibitors of NaCl influx, elicited delta psi a,b and decreased the transepithelial potential (psi t) in parallel. Regardless of initial values, psi a and psi b approached the equilibrium potential for K (EK) so that in the steady state following inhibition of NaCl entry, psi a approximately equal to psi b approximately equal to ECl approximately equal to EK. Bumetanide decreased cell Cl activity (aClc) toward equilibrium levels. Bumetanide and cGMP decreased the fractional apical membrane resistance (fRa), increased the slope of the relation of psi a to [K]m, and decreased cellular conductance (Gc) by approximately 85%, which indicates a marked increase in basolateral membrane conductance (Gb). Since the basolateral membrane normally shows a high conductance to Cl, a direct relation between apical salt entry and GClb is suggested by these findings. As judged by the response to bumetanide or ion replacement in the presence of mucosal Ba, inhibition of Na/K/Cl co-transport alone is not sufficient to elicit delta psi a,b. This suggests the presence of a parallel NaCl co-transport mechanism that may be activated when Na/K/Cl co-transport is compromised. The delta psi a,b response to reduced apical NaCl entry would assist in maintaining the driving force for Na-coupled amino acid uptake across the apical membrane as luminal [NaCl] falls during absorption.

Animals↗
PubMed1984

Potassium transport by turtle colon: active secretion and active absorption.

Measurement of transepithelial potassium fluxes in the absence of transmural electrochemical potential gradients showed that the isolated turtle colon can actively absorb and actively secrete K+. Under short-circuit conditions the active secretory flow was inhibited by mucosal amiloride, whereas the absorptive flow was unaffected by the diuretic. The effects of ouabain and barium on secretory flow were consistent with a simple model involving basolateral uptake by an Na+-K+-ATPase and conductive exist across the apical and basolateral membranes. The active absorptive flux was blocked by mucosal ouabain and by serosal barium. The opposing active flows clearly represented cellular K+ transport, whereas paracellular K+ flows behaved as expected for a free-solution shunt.

Absorption↗
PubMed1984

Control of potassium transport by turtle colon: role of membrane potential.

To more clearly define the role of the transepithelial electrical potential difference (V m----s), potassium permeability, and sodium-potassium pump rate in transcellular potassium transport by isolated turtle colon, we measured transmural potassium fluxes under open-circuit conditions in the presence and absence of putative blockers of potassium transport: amiloride and barium. The results were consistent with the notion that V m----s is a major determinant of cellular potassium secretion, whereas active potassium absorption is insensitive to changes in V m----s. These observations suggest that "coupling" between colonic sodium absorption and potassium secretion in vivo could be due primarily to the effect of the lumen negative V m----s on transcellular secretory potassium flow. Amiloride-induced inhibition of potassium secretion appeared to be due to the reductions in V m----s and sodium-potassium pump rate that accompanied the inhibition of active sodium absorption.

Amiloride↗
PubMed1984

Potassium transport by flounder intestinal mucosa.

We studied the mechanisms of K transport across an epithelium in which NaCl absorption is mediated primarily by Na/K/Cl cotransport at the apical membrane. Rubidium served as a reliable K substitute; under control conditions, both K and Rb were actively secreted. During secretion, K (Rb) enters across the basolateral membrane via the Na/K pump and exits across the apical membrane through K conductance pathways, since serosal ouabain or mucosal barium abolished K secretion, mucosal furosemide or Cl-free media blocked K secretion by interfering with access of Na to the pump, and elevated mucosal solution [K] or [Rb] depolarized the apical membrane electrical potential difference. Mucosal Ba unmasked active Rb absorption that could be blocked by mucosal furosemide. These findings illustrate active K absorption and secretion across an epithelium that comprises a single cell type in which opposing K fluxes across the apical membrane are mediated by Na/K/Cl cotransport entry and conductive K exit. The direction of transepithelial K transport is determined by the relative activities of these pathways.

Animals↗
PubMed1980

Active sodium transport by turtle colon via an electrogenic Na-K exchange pump.

Active sodium absorption by a variety of epithelia is abolished by ouabain, but the obligatory coupling between the movement of sodium and potassium expected from a basolateral (Na+ + K+) ATPase has not been convincingly demonstrated. According to the model of Koefoed-Johnsen and Ussing, the asymmetric cation selectivities of the apical and basolateral membranes prevent basolateral Na-K exchange from being expressed as opposing transmural ion flows. An additional consequence of this asymmetry is that the short-circuit current (Isc) cannot be identified with the current through the sodium-potassium pump. We used the polyene antibiotic, amphotericin-B, to reduce the resistance and the cation selectivity of the apical membrane of isolated turtle colon so that the basolateral membrane current could be dissected into two components: one through a barium-sensitive potassium channel and another which represents the current associated with ouabain-sensitive, electrogenic, Na-K exchange. Comparison of cation fluxes and short circuit current indicates that in these conditions active sodium absorption is entirely attributable to an electrogenic Na-K pump with a stoichiometry of approximately 3Na:2K.

Amphotericin B↗
PubMedDate not supplied

Phosphorylation fails to activate chloride channels from cystic fibrosis airway cells.

Chloride impermeability of epithelial cells can account for many of the experimental and clinical manifestations of cystic fibrosis (CF). Activation of apical-membrane Cl- channels by cyclic AMP-mediated stimuli is defective in CF airway epithelial cells, despite normal agonist-induced increases in cellular cAMP levels. This defect in Cl- channel regulation has been localized to the apical membrane by exposing the cytoplasmic surface of excised membrane patches to the catalytic subunit (C subunit) of cAMP-dependent protein kinase and ATP. In membranes from normal cells, C-subunit activated Cl- channels with properties identical to those stimulated by cAMP-dependent agonists during cell-attached recording. Activation by the C subunit was not observed in CF membranes, but the presence of Cl- channels was verified by voltage-induced activation. The failure of the C subunit to activate the Cl- channels of CF membranes indicates that the block in their cAMP-mediated activation lies distal to induction of cAMP-dependent protein kinase activity and focuses our attention on the Cl- channel and its membrane-associated regulatory proteins as the probable site of the CF defect.

Cell Membrane↗