Vibrating probe analysis of teleost opercular epithelium: correlation between active transport and leak pathways of individual chloride cells.
We have utilized the vibrating probe technique to examine transport by individual chloride cells in the short-circuited fish opercular epithelium. Variability in the steady state and in response to rapid perturbations, including fast-acting hormones and ion replacement, was analyzed. Negative short-circuit currents, corresponding to chloride secretion, were associated with the apical crypts of all but five of 386 chloride cells sampled. Average chloride cell short-circuit current and conductance were 2.7 +/- 0.1 nA and 87.7 +/- 3.8 nS, respectively, or 19 mA cm-2 and 620 mS cm-2 (resistance = 1.6 omega cm2) when normalized to apical crypt surface area. Exposure to 1 microM epinephrine rapidly inhibited the tissue short-circuit current by inhibiting the current pumped by all chloride cells, i.e. all chloride cells have adrenergic receptors. The time course of inhibition for each cell mirrored that of the whole tissue. Reversal of epinephrine inhibition of the tissue short-circuit current by glucagon and phosphodiesterase inhibition was by reversal of epinephrine's inhibition of individual chloride cells, and not by turning on cells which were previously inactive or uninhibited, or by stimulating nonchloride cells. A great amount of variability existed among chloride cells in the ability of these agents to reverse epinephrine-inhibited current. Likewise, considerable variability in the response of chloride cell conductance to these perturbations was observed, and in many instances a clear dissociation between current and conductance was noted. In the steady state, variability among cells in a single tissue always defined a linear relationship between chloride cell current and conductance with zero-current conductance intercept at zero. Equivalent circuit modeling indicates that the leak conductance of chloride cells within a single tissue always contributes the same proportion to the total individual chloride cell conductance, such that the ratio between the conductances of the active and leak pathways of chloride cells is constant. The leak pathway is almost certainly dominated by a sodium-selective paracellular pathway. The results suggest that these cells control the permeability of their paracellular pathway. A possible mechanism for this control is discussed.
J K
JK
Foskett
T E
TE
Machen
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D002712:Chlorides / Q000378:metabolism*; D004553:Electric Conductivity / Q000187:drug effects; D004837:Epinephrine / Q000494:pharmacology; D004848:Epithelium / Q000502:physiology* / Q000648:ultrastructure; D005399:Fishes; D007700:Kinetics; D008433:Mathematics; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D014732:Vibration
D002712:Chlorides; D015056:1-Methyl-3-isobutylxanthine; D004837:Epinephrine