Cationic amphiphilic drugs inhibit the internalization of cholera toxin to the Golgi apparatus and the subsequent elevation of cyclic AMP.
Cholera toxin (CT) consists of a pentameric B subunit which binds with high affinity to ganglioside GM1, and an A subunit which stimulates adenylate cyclase, resulting in the elevation of cAMP. We now examine the effect of cationic amphiphilic drugs (CADs) on the internalization of rhodamine (Rh)-CT in cultured hippocampal neurons. CADs have recently been shown to inhibit receptor recycling by disrupting the assembly-disassembly of clathrin at the plasma membrane and on endosomes (Wang, L.-H., Rothberg, K. G., and Anderson, R. G. W. (1993) J. Cell Biol. 123, 1107-1117). Rh-CT was internalized by an energy- and temperature-dependent (presumably vesicular) mechanism to the Golgi apparatus. Internalization to the Golgi apparatus was completely but reversibly blocked by CADs, and the ability of CT to stimulate the elevation of cAMP was significantly reduced. In control cells, cAMP levels were elevated 2.3-fold after 20 min of incubation with CT, but in CAD-treated cells cAMP levels were only elevated 1.3-fold. The effect of CADs on CT internalization was not due to a direct effect of CADs on the Golgi apparatus. Our data demonstrate that CADs inhibit vesicular transport of CT to the Golgi apparatus and imply that the sorting of CT to the Golgi apparatus occurs in the same endosomal compartment involved in sorting recycling receptors to the plasma membrane, since both pathways are inhibited by CADs.
Sofer
A
A
Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Futerman
A H
AH
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002478:Cells, Cultured; D002746:Chlorpromazine / Q000494:pharmacology*; D002772:Cholera Toxin / Q000378:metabolism*; D000242:Cyclic AMP / Q000096:biosynthesis*; D003864:Depression, Chemical; D004705:Endocytosis / Q000187:drug effects*; D037341:Fumonisins; D005677:G(M1) Ganglioside / Q000378:metabolism / Q000494:pharmacology; D006056:Golgi Apparatus / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology; D007099:Imipramine / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D009183:Mycotoxins / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012235:Rhodamines / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013110:Sphingosine / Q000494:pharmacology*
D037341:Fumonisins; D009183:Mycotoxins; D012235:Rhodamines; D005677:G(M1) Ganglioside; C056933:fumonisin B1; D002772:Cholera Toxin; D000242:Cyclic AMP; D013110:Sphingosine; D007099:Imipramine; D002746:Chlorpromazine