Summary
We have adapted the vibrating probe extracellular recording technique to use on an epithelium under voltage clamp in an Ussing chamber. The vibrating probe allows very low drift measurements of current density immediately over the epithelial surface. These measurements allowed sites of electrogenic transport in the epithelium to be localized with a spatial resolution of 5 μm. The technique was applied to the opercular membrane of the teleost fish, the tilapia,Sarotherodon mossambicus. The mitochondrion-rich “chloride cells” were shown to be the only sites of electrogenic ion transport in this heterogeneous epithelium. Cell sampling experiments demonstrated variable negative short-circuit currents associated with nearly all of approximately 300 chloride cells examined, which appeared to account for all of the tissue shortcircuit current. Current-voltage relations for individual cells were also measured. Conductance associated with chloride cells (i.e. cellular and junctional pathways) accounted for all but 0.5 mS/cm2 of the tissue conductance, with the balance apparently accounted for by leak pathways near the edge of the tissue. Current and conductance associated with other cell types was at least 50-fold smaller than for the chloride cell. Chloride-free solutions reduced chloride cell current and conductance by 98 and 95%, respectively.
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Bobrycki, V.A., Mills, J.W., Macknight, A.D.C., DiBona, D.R. 1981. Structural responses to voltage clamping in the toad urinary bladder. I. The principal role of granular cells in the active transport of sodium.J. Membrane Biol. 60:21–33
Cereijido, M., Stefani, E., Martinez-Palomo, A. 1980. Occluding junctions in a cultured transporting epithelium: Structural and functional heterogeneity.J. Membrane Biol. 53:19–32
Davenport, W.B. 1970. Probability and Random Processes. p. 518. McGraw-Hill, New York
DiBona, D.R., Sherman, B., Bobrycki, V.A., Mills, J.W., Macknight, A.D.C. 1981. Structural response to voltageclamping in the toad urinary bladder. II. Granular cells and the natriferic action of vasopressin.J. Membrane Biol 60:35–44
Foskett, J.K., Bern, H.A., Machen, T.E., Connor, M. 1983. Chloride cells and the hormonal control of teleost fish osmoregulation.J. Exp. Biol. Rev. 5:(in press)
Foskett, J.K., Logsdon, D., Turner, T., Machen, T.E., Bern, H.A. 1981. Differentiation of the chloride extrusion mechanism during seawater adaptation of a teleost fish, the cichlidSarotherodon mossambicus.J. Exp. Biol. 93:209–224
Foskett, J.K., Machen, T.E., Bern, H.A. 1982. Chloride secretion and conductance of teleost opercular membrane: Effects of prolactin.Am. J. Physiol. 242:R380-R389
Foskett, J.K., Scheffey, C. 1982. The chloride cell: Definitive identification as the salt-secretory cell in teleosts.Science 215:164–166
Frömter, E. 1972. The route of passive ion movement through the epithelium ofNecturus gallbladder.J. Membrane Biol. 8:259–301
Frömter, E., Diamond, J. 1972. Route of passive ion permeation in epithelia.Nature, New Biol. 235:9–13
Handler, J.S., Perkins, F.M., Johnson, J.P. 1980. Studies of renal cell function using cell culture techniques.Am. J. Physiol. 238:F1-F9
Hudspeth, A.J. 1975. Establishment of tight junctions between epithelial cells.Proc. Natl. Acad. Sci. USA 72:2711–2713
Husted, R.F., Mueller, A.L., Kessel, R.G., Steinmetz, P.R. 1981. Surface characteristics of carbonic-anhydrase-rich cells in the turtle urinary bladder.Kidney Int. 19:491–502
Jaffe, L.F. 1981. Control of development by steady ionic currents.Fed. Proc. 40:125–127
Jaffe, L.F., Nuccitelli, R. 1974. An ultrasensitive vibrating probe for measuring steady extracellular currents.J. Cell Biol. 63:614–628
Jaffe, L.F., Nuccitelli, R. 1977. Electrical controls of development.Annu. Rev. Biophys. Bioeng. 6:445–476
Jarrell, J.A., King, J.G., Mills, J.W. 1981. A scanning micropipette molecule microscope.Science 211:277–279
Karnaky, K.J. 1980. Ion-secreting epithelia: Chloride cells in the head region ofFundulus heteroclitus.Am. J. Physiol. 238:R185-R198
Lettir, M., Kaissling, B., Koeppen, B.M., Wade, J.B. 1982. Binding of peanut lectin to specific epithelial cell types in kidney.Am. J. Physiol. 242:C117-C120
Marshall, W.S., Nishioka, R.S. 1980. Relation of mitochondria-rich chloride cells to active chloride transport in the skin of a marine teleost.J. Exp. Zool. 214:147–188
Mills, J.W., Malick, L.E. 1978. Mucosal surface morphology of the toad urinary bladder. Scanning electron microscope study of the natriferic and hydro-osmotic response to vasopressin.J. Cell Biol. 77:598–610
Narbaitz, R., Kacew, S., Sitwell, L. 1981. Carbonic anhydrase activity in the chick embryo chorioallantois: Regional distribution and vitamin D regulation.J. Embryol. Exp. Morphol. 65:127–137
Rosen, S., Oliver, J.A., Steinmetz, P.R. 1974. Urinary acidification and carbonic anhydrase distribution in bladders of Dominican and Columbian toads.J. Membrane Biol 15:193–205
Scott, W.N., Sapirstein, V.S., Yoder, M.J. 1974. Partition of tissue functions in epithelia: Localization of enzymes in “mitochondrion-rich” cells of toad urinary bladder.Science 184:797–800
Stetson, D.L., Wade, J.B., Giebisch, G. 1980. Morphologic alterations in the rat medullary collecting duct following potassium depletion.Kidney Int. 17:45–56
Ussing, H.H. 1949. The active ion transport through the isolated frog skin in the light of tracer studies.Acta Physiol. Scand. 17:1–17
Voûte, C.L., Meier, W. 1978. The mitochondria-rich cell of frog skin as hormone-sensitive “shunt-path.”.J. Membrane Biol. Special Issue:151–165
Wade, J.B., O'Neil, R.G., Pryor, J.L., Boulpaep, E.L. 1979. Modulation of cell membrane area in renal collecting tubule by corticosteroid hormones.J. Cell Biol. 81:439–445
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Scheffey, C., Foskett, J.K. & Machen, T.E. Localization of ionic pathways in the teleost opercular membrane by extracellular recording with a vibrating probe. J. Membrain Biol. 75, 193–203 (1983). https://doi.org/10.1007/BF01871950
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DOI: https://doi.org/10.1007/BF01871950
