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Injection of RNA from carp retina induces the formation of a membrane potassium channel in Xenopus oocytes

Published online by Cambridge University Press:  02 June 2009

Lawrence H. Pinto
Affiliation:
Department of Neurobiology and Physiology, Northwestern Univeristy, Evanston
Akimichi Kaneko
Affiliation:
National Institute for Physiological Sciences, Myodaiji 444 Okazaki, Japan

Abstract

Total RNA was purified from freshly isolated retinas of adult carp and injected into oocytes of Xenopus laevis (stage 5–6). Two to six days after injection, depolarizing voltage-clamp steps evoked a slowly activated outward currents as large as 3 μA. This current inactivated slowly with a single time constant (τ= 3.1 ± 0.24 S.E.M., for Vm= +30 mV). The current was inhibited by tetraethylammonium (3.8 mM for half-maximal inhibition). In the presence of Co2+ (1 mM) or barium methanesulfonate (40 mM), the current-voltage relationship shifted to slightly more depolarized values (5–10 mV); the maximal value of the current that was sensitive to Co2+ or Ba2+ treatments was only a small fraction (about 10%) of the TEA-sensitive current, and its current-voltage relationship was similar to that for uninjected oocytes. The reversal potential of the membrane current was studied with [K+]0 of 1–77 mM. For [K+]0 > 20 mM, the reversal potential changed with a slope of 63 mV (±;2 mV S.E.M.) per 10-fold change in [K+]0. The conductance was induced half-maximally at 17 mV (±;0.9 mV s.e.m.). The depolarization required for an e−fold increase in conductance was 13 mV (±;0.6 mV s.e.m.). From these results, we conclude that the injection of total RNA from carp retinas induces the formation of a membrane K+ channel in Xenopus oocytes. The channel formed has many of properties reported for the maintained outward current of goldfish horizontal and bipolar cells.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1991

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References

Bader, C.R., Bertrand, D. & Schwartz, E.A. (1982). Voltage-activated and calcium-activated currents studies in solitary rod inner segments from the salamander retina. Journal of Physiology 331, 253284.CrossRefGoogle Scholar
Barish, M.E. (1983). A transient calcium-dependent chloride current in the immature Xenopus oocyte. Journal of Physiology 342, 309325.CrossRefGoogle ScholarPubMed
Baylor, D., Fuortes, M. & O'Bryan, P.M. (1971). Receptive fields of cones in the retina of the turtle. Journal of Physiology 214, 265294.CrossRefGoogle ScholarPubMed
Dascal, N., Snutch, T.P., Lubbert, H., Davidson, N. & Lester, H.A. (1986). Expression and modulation of voltage-gated calcium channels after RNA injection in Xenopus oocytes. Science 231, 11471150.CrossRefGoogle ScholarPubMed
Eliasof, S., Barnes, S. & Werblin, F. (1987). The interaction of ionic currents mediating single spike activity in retinal amacrine cells of the tiger salamander. Journal of Neuroscience 7(11), 35123524.CrossRefGoogle ScholarPubMed
Frech, G.C., Van, Dongen A.M.J., Schuster, G., Brown, A.M. & Joho, R.H. (1989). A novel potassium channel with delayed rectifier properties isolated from rat brain by expression cloning. Nature 340, 642645.CrossRefGoogle ScholarPubMed
Hille, B. (1984). Ionic Channels of Excitable Membranes. Sunderland; Sinauer, pp. 100108.Google Scholar
Kaneko, A., Pinto, L.H. & Tachibana, M. (1989). Transient calcium current of retinal bipolar cells of the mouse. Journal of Physiology 410, 613629.CrossRefGoogle ScholarPubMed
Kaneko, A. & Shimazaki, H. (1976). Synaptic transmission from photoreceptors to bipolar and horizontal cells in the carp retina. Cold Spring Harbor Symposia on Quantitative Biology 40, 537546.CrossRefGoogle ScholarPubMed
Kaneko, A. & Tachibana, M. (1985). A voltage-clamp analysis of membrane currents in solitary bipolar cells dissociated from Carassius auratus. Journal of Physiology 358, 131152.CrossRefGoogle ScholarPubMed
Katz, B. (1966). Nerve, Muscle, and Synapse. New York: McGraw Hill, p. 193.Google Scholar
Kuffler, S.W., Nicholls, J.G. & Martin, A.R. (1984). A Cellular Approach to the Function of the Nervous System. Sunderland: Sinauer, p. 651.Google Scholar
Leonard, J.P., Nargeot, J., Snutch, T.P., Davidson, N. & Lester, H.A. (1987). Ca channels induced in Xenopus oocytes by rat brain mRNA. Journal of Neuroscience 7(3), 875881.CrossRefGoogle ScholarPubMed
Lipton, S.A. & Tauck, D.L. (1987). Voltage-dependent conductances of solitary ganglion cells dissociated from the rat retina. Journal of Physiology 385, 361391.CrossRefGoogle ScholarPubMed
Miledi, R. (1982). A calcium-dependent transient outward current in Xenopus laevis oocytes. Proceedings of the Royal Society (London) 215, 491497.Google ScholarPubMed
Murakami, M. & Shimoda, Y. (1977). Identification of amacrine and ganglion cells in the carp retina. Journal of Physiology 264, 801818.CrossRefGoogle ScholarPubMed
Tachibana, M. (1983). Ionic currents of solitary horizontal cells isolated from goldfish retina. Journal of Physiology 345, 239351.CrossRefGoogle ScholarPubMed
Werblin, F.S. & Dowling, J.E. (1969). Organization of the retina of the mudpuppy, Necturus maculosus, II: Intracellular recording. Journal of Neurophysiology 32, 339355.CrossRefGoogle ScholarPubMed
White, M.M. & Aylwin, M. (1990). Niflumic and Flufenamic acids are potent reversible blockers of Ca2+-activated Cl channels in Xenopus oocytes. Molecular Pharmacology 37, 720724.Google ScholarPubMed