The polarized distribution of poly(A+)-mRNA-induced functional ion channels in the Xenopus oocyte plasma membrane is prevented by anticytoskeletal drugs.
Foreign mRNA was expressed in Xenopus laevis oocytes. Newly expressed ion currents localized in defined plasma membrane areas were measured using the two-electrode voltage clamp technique in combination with a specially designed chamber, that exposed only part of the surface on the oocytes to channel agonists or inhibitors. Newly expressed currents were found to be unequally distributed in the surface membrane of the oocyte. This asymmetry was most pronounced during the early phase of expression, when channels could almost exclusively be detected in the animal hemisphere of the oocyte. 4 d after injection of the mRNA, or later, channels could be found at a threefold higher density at the animal than at the vegetal pole area. The pattern of distribution was observed to be similar with various ion channels expressed from crude tissue mRNA and from cRNAs coding for rat GABAA receptor channel subunits. Electron microscopical analysis revealed very similar microvilli patterns at both oocyte pole areas. Thus, the asymmetric current distribution is not due to asymmetric surface structure. Upon incubation during the expression period in either colchicine or cytochalasin D, the current density was found to be equal in both pole areas. The inactive control substance beta-lumicolchicine had no effect on the asymmetry of distribution. Colchicine was without effect on the amplitude of the expressed whole cell current. Our measurements reveal a pathway for plasma membrane protein expression endogenous to the Xenopus oocyte, that may contribute to the formation and maintenance of polarity of this highly organized cell.
Peter
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AB
Department of Pharmacology, University of Bern, Switzerland.
Schittny
J C
JC
Niggli
V
V
Reuter
H
H
Sigel
E
E
D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D002478:Cells, Cultured; D003078:Colchicine / Q000494:pharmacology*; D015638:Cytochalasin D / Q000494:pharmacology*; D007473:Ion Channels / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008854:Microscopy, Electron; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D011061:Poly A / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D015222:Sodium Channels / Q000378:metabolism; D013499:Surface Properties; D014982:Xenopus laevis; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D007473:Ion Channels; D012333:RNA, Messenger; D015222:Sodium Channels; D015638:Cytochalasin D; D011061:Poly A; D005680:gamma-Aminobutyric Acid; D003078:Colchicine