Interaction of the B subunit of cholera toxin with endogenous ganglioside GM1 causes changes in membrane potential of rat thymocytes.
The fluorescent anionic dye, bisoxonol, and flow cytometry have been used to monitor changes in the membrane potential of rat thymocytes exposed to the B subunit of cholera toxin. The B subunit induced a rapid hyperpolarization, which was due to activation of a Ca2+-sensitive K+ channel. Reduction of extracellular Ca2+ to less than 1 microM by the addition of [ethylene-bis(oxyethylenenitrilo)]tetraacetic acid immediately abolished the hyperpolarization caused by the B subunit. Cells treated with quinine and tetraethylammonium lost their ability to respond to the B subunit, whereas 4-aminopyridine did not have any effect. Thus, calcium-sensitive and not voltage-gated K+ channels appeared to be responsible for the hyperpolarization. The results of ion substitution experiments indicated that extracellular Na+ was not essential for changes in membrane potential. Further studies with ouabain, amiloride and furosemide demonstrated that electrogenic Na+/K+ ATPase, Na+/H+ antiporter and Na+/K+/Cl- cotransporter, respectively, were not involved in the hyperpolarization process induced by the B subunit. Thus, crosslinking of several molecules of ganglioside GM1 on the cell surface of rat thymocytes by the pentavalent B subunit of cholera toxin modulated plasma membrane permeability to K+ by triggering the opening of Ca2+-sensitive K+ channels. A role for gangliosides in regulating ion permeability would have important implications for the function of gangliosides in various cellular phenomena.
Division of Nutrition, Food and Drug Administration, Washington, D.C. 20204.
S A
SA
Mulhern
P H
PH
Fishman
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S
Spiegel
D000818:Animals; D002118:Calcium / Q000502:physiology*; D002772:Cholera Toxin / Q000378:metabolism*; D003208:Concanavalin A / Q000494:pharmacology; D005434:Flow Cytometry; D005677:G(M1) Ganglioside / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D013950:Thymus Gland / Q000378:metabolism* / Q000502:physiology
D015221:Potassium Channels; D003208:Concanavalin A; D005677:G(M1) Ganglioside; D002772:Cholera Toxin; D002118:Calcium