Summary
The cellular mechanism of K-stimulated Cl transport in locust hindgut was studied using double-barrelled ionsensitive microelectrodes and electrophysiological techniques. Steady-state net electrochemical potentials for Cl and K and the conductances of apical and basal membranes and paracellular pathway were determined under control conditions, during exposure to 1mm cAMP, and following ion substitutions. Under control open-circuit conditions, intracellular Cl activity (a cCl ) was 3.5 times that predicted for passive equilibrium across the apical membrane. The net electrochemical potential opposing Cl entry from the mucosal side\((\Delta \bar \mu _{Cl}^a /F)\) increased by 50% during cAMP stimulation of transepithelial Cl absorption whereas the net electrochemical potential favoring Cl exit across the basal membrane\(( - \Delta \bar \mu _{Cl}^b /F)\) was unchanged. No correlation was observed between\(\Delta \bar \mu _{Cl}^a /F\) and the net electrochemical potential across the apical membrane for Na. The net electrochemical potential favoring K entry across the apical membrane\(( - \Delta \bar \mu _K^a /F)\) was negligible underI sc conditions when Cl transport rate was approximately 10 μeq cm−2 hr−1. Locust rectal cells showed electrical and dye coupling. The results also indicate that most transepithelial diffusion of ions is transcellular and that epithelial tightness effectively increases during exposure to cAMP becauseR a andR b both decrease, by ≈80% whileR j is unchanged. The cAMP-induced δR b was abolished in Cl-free saline whereas δR a was insensitive to Cl removal, but was blocked by removing K from the saline. Based on these findings, our model for Cl absorption in locust hindgut features i) an active entry step for Cl at the apical membrane which is stimulated by cAMP and by low levels of K on the mucosal side, but is not energized by\( - \Delta \bar \mu _{Na}^a /F\) or\( - \Delta \bar \mu _K^a /F\) a large cAMP-stimulated Cl conductance in the basal membrane and a similar cAMP-stimulated K conductance in the apical membrane. cAMP dose-response curves are similar for the stimulation of active Cl absorption and Cl-independent (i. e. K) conductance, indicating that cAMP exerts dual control over active Cl transport and counter-ion permeability.
Similar content being viewed by others
References
Augustus, J., Bijman, J., Os, C.H. van 1978. Electrical resistance of rabbit submaxillary main duct: A tight epithelium with leaky cell membranes.J. Membrane Biol. 43:203–226
Baumgarten, C.M., Fozzard, H.A. 1981. Intracellular chloride activity in mammalian ventricular muscle.Am. J. Physiol. 241:C121-C129
Berridge, M.J. 1981. Hormone-induced changes in ion level during stimulation of fluid secretion by gland cells.In The Application of Ion-selective Microelectrodes. T. Zeuthen, editor. pp. 61–74. North-Holland Biomedical, New York
Berridge, M.J., Lindley, B.D., Prince, W.T. 1975. Membrane permeability changes during stimulation of isolated salivary glands ofCalliphora by 5-hydroxytryptamine.J. Physiol. (London) 244:549–567
Berridge, M.J., Schlue, W.R. 1978. Ion-selective electrode studies on the effects of 5-hydroxytryptamine on the intracellular level of potassium in an insect salivary gland.J. Exp. Biol. 72:203–216
Blankemeyer, J.T., Duncan, R.L. 1980. The potassium activity in a polymorphic potassium active transporting epithelium, insect midgut.Fed. Proc. 39:1711
Boulpaep, E.L., Sackin, H. 1980. Electrical analysis of intraepithelial barriers.In: Current Topics in Membranes and Transport. F. Bronner and A. Kleinzeller, editors., Vol. 13, pp. 169–197. Academic, New York
Cereijido, M., Borboa, L., Gonzalez-Mariscal, L. 1983. Occluding junctions and paracellular pathway in monolayers of MDCK cells.J. Exp. Biol. 106:205–215
Clausen, C., Machen, T.E., Diamond, J. 1983. Use of AC impedance analysis to study membrane changes related to acid secretion in amphibian gastric mucosa.Biophys. J. 41:167–178
Delong, J., Civan, M.M. 1978. Dissociation of cellular K+ accumulation from net Na+ transport by toad urinary bladder.J. Membrane Biol. 42:19–43
Duffey, M.E., Turnheim, K., Frizzell, R.A., Schultz, S.G. 1978. Imtracellular chloride activities in rabbit gallbladder: Direct evidence for the role of the sodium-gradient in energizing “uphill” chloride transport.J. Membrane Biol. 42:229–245
Eisenberg, R.S., Johnson, E.A. 1970. Three-dimensional electrical field problems in physiology.In: Progress in biophysics and molecular biology J.A.V. Butler and D. Noble, editors. Vol. 20. pp. 1–65. Pergamon, Toronto
Frömter, E. 1972. The route of passive ion movement through the epithelium ofNecturus gallbladder.J. Membrane Biol. 8:259–301
Fujimoto, M., Kotera, K., Matsumura, Y. 1980. The direct measurement of K, Cl, Na and H ions in bullfrog tubule cells.In: Current Topics in Membranes and Transport. F. Bronner and A. Kleinzeller, editors. Vol. 13, pp. 49–61. Academic, New York
Fujimoto, M., Kubota, T. 1976. Physicochemical properties of a liquid ion exchanger microelectrode and its application to biological fluid.Jpn. J. Physiol. 26:631–650
Greger, R. 1981. Chloride reabsorption in the rabbit cortical thick ascending limb of Henle's loop of rabbit kidney.Pfluegers Arch. 390:30–37
Guggino, W.B., Windhager, E.E., Boulpaep, E.L., Giebisch, G. 1982. Cellular and paracellular resistances of theNecturus proximal tubule.J. Membrane Biol. 67:143–154
Gupta, B.L., Wall, B.J., Oschman, J.L., Hall, T.A. 1980. Direct microprobe evidence of local concentration gradients and recycling of electrolytes during fluid absorption in the rectal papillae ofCalliphora.J. Exp. Biol. 88:21–47
Hanrahan, J.W. 1982. Cellular mechanism and regulation of KCl transport across an insect epithelium. Ph. D. Thesis. University of British Columbia, Vancouver
Hanraban, J.W., Meredith, J., Phillips, J.E., Brandys, D. 1983. Methods for the study of transport and control in insect hindgut.In: Measurement of Ion Transport and Metabolic Rate in Insects. T. Bradley and T. Miller, editors. pp. 19–67. Springer-Verlag, New York
Hanrahan, J.W., Phillips, J.E. 1982. Electrogenic, K+-dependent chloride transport in locust hindgut.Philos. Trans. R. Soc. London B 299:585–595
Hanrahan, J.W., Phillips, J.E. 1983. K and cAMP stimulate the Cl pump in locust rectum.Am. Zool. 22:914
Hanrahan, J.W., Phillips, J.E. 1983. Mechanism and control of salt absorption in locust rectum.Am J. Physiol. 224:R131-R142
Hanrahan, J.W., Wills, N.K., Lewis, S.A. 1983. Bariuminduced current fluctuations from the basal membrane of an insect epithelium.Proc. Int. Congr. Physiol. Sci. p. 457
Herrera, L., Lopes-Moratalla, N., Santiago, E., Ponz, F., Jordana, R. 1978. Effect of bicarbonate on chloride-dependent transmural potential and ATPase activity in the rectal wall ofSchistocerca gregaria.Rev. Esp. Fisiol. 34:219–224
Komnick, H., Schmitz, M.H., Hinssen, H. 1980. Biochemischer Nachweis von HCO −3 - und Cl−-abhängigen ATPase-activitäten im rectum von anisopteranen Libellenlarven und Hemmung der rectalen chiloridaufnahme durch thiocyanat.Eur. J. Cell. Biol.,20:217–227
Kotera, K., Satake, N., Honda, M., Fujimoto, M. 1979. The measurement of intracellular sodium activities in the bullfrog by means of a double-barrelled sodium ion-exchange microelectrode.Membr. Biochem. 2:323–338
Lewis, S.A., Eaton, D.C., Diamond, J.M. 1976. The mechanism of Na+ transport by rabbit urinary bladder.J. Membrane Biol. 28:41–70
Loewenstein, W.R. 1981. Junctional intercellular communication: The cell-to-cell membrane channel.Physiol. Rev. 61:829–913
Marshall, W.S., Klyce, S.D. 1983. Cellular and paracellular pathway resistances in the “tight” Cl−-secreting epithelium of rabbit cornea.J. Membrane Biol. 73:275–282
Moffett, D.F., Hudson, R.L., Moffett, S.B., Ridgway, R.L. 1982. Intracellular K+ activities and cell membrane potentials in a K+-transporting epithelium, the midgut of tobacco hornworm(Manduca sexta).J. Membrane Biol. 70:59–68
Ogden, T.E., Citron, M.C., Pierantoni, R. 1978. The jet stream microbeveler: An inexpensive way to bevel ultrafine glass micropipettes.Science 201:469–470
Olver, F.W.J. 1967. Bessel functions of integer order.In: Handbook of Mathematical Functions. M. Abramowitz and J.A. Stegun, editors. pp. 355–422. National Bureau of Standards, Washington, D.C
Phillips, J.E. 1964. Rectal absorption in the desert locustSchistocerca gregaria Forskål. II. Sodium, potassium and chloride.J. Exp. Biol. 41:39–67
Phillips, J. 1970 Apparent transport of water by insect excretory systems.Am. Zool. 10:413–436
Reuss, L., Finn, A.L. 1974. Passive electrical properties of toad urinary bladder epithelium. Intercellular electrical coupling and transepithelial cellular and shunt conductances.J. Gen. Physiol. 64:1–25
Reuss, L., Finn, A.L. 1977. Mechanisms of voltage transients during current clamp inNecturus gallbladder.J. Membrane Biol. 37:299–319
Reuss, L., Weinman, S.A. 1979. Intracellular ionic, activities and transmembrane electrochemical potential differences in gallbladder epithelium.J. Membrane Biol. 49:345–362
Robinson, R.A., Stokes, R.H. 1970. Electrolyte solutions (Wnd ed., revised). Butterworths, London
Shiba, H. 1971. Heaviside's “Bessel cable” as an electric model for flat simple epithelial cells with low resistive junctional membranes.J. Theor. Biol. 30:59–68
Socolar, S.J., Politoff, A.L. 1971. Uncoupling cell junctions of a glandular epithelium by depolarizing current.Science 172:492–494
Spenney, J.G., Shoemaker, R.L., Sachs, G. 1974. Microelectrode studies of fundic gastric mucosa: Cellular coupling and shunt conductance.J. Membrane Biol. 19:105–128
Spring, J.H., Phillips, J.E. 1980. Studies on locust rectum: II. Identification of specific ion transport processes regulated by corpora cardiacum and cyclic-AMP.J. Exp. Biol. 86:225–236
Spring, K.R., Kimura, G. 1978. Chloride reabsorption by renal proximal tubules ofNecturus.J. Membrane Biol. 38:233–254
Stewart, W.W. 1978. Functional connections between cells as revealed by dye-coupling with a highly fluorescent naphthalimide tracer.Cell 14:741–759
Vietinghoff, U.E., Olszewska, E., Janiszewski, L. 1969. Measurement of the biolectric potentials in the rectum ofLocusta migratoria andCarausius morosus inin vitro preparations.J. Insect Physiol. 15:1273–1277
Walker, J.L. 1971. Ion specific liquid ion exchanger microelectrodes.Anal. Chem. 43:89A-92A
Williams, D., Phillips, J., Prince, W., Meredith, J. 1978. The source of short-circuit current across locust rectum.J. Exp. Biol. 77:107–122
Williams, J.C., Jr. 1983. The Malpighian tubule of the yellow fever mosquito: Its functionin vitro andin vivo. Ph.D. Thesis. Cornell University, Ithaca
Wood, J.L., Moreton, R.B. 1978. Refinements in the shortcircuit technique, and its application to active potassium transport across theCecropia midgut.J. Exp. Biol. 77:123–140
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hanrahan, J.W., Phillips, J.E. KCl transport across an insect epithelium: II. Electrochemical potentials and electrophysiology. J. Membrain Biol. 80, 27–47 (1984). https://doi.org/10.1007/BF01868688
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01868688