Calcium wave evoked by activation of endogenous or exogenously expressed receptors in Xenopus oocytes.
The mRNA encoding the cloned substance K receptor was microinjected into Xenopus laevis oocytes. After expression of the mRNA, Ca2+ was imaged in the oocytes with a digital imaging fluorescence microscopy system using the Ca2(+)-sensitive dyes fura-2 and fluo-3. Application of substance K caused a dose-related wave of Ca2+ mobilization to spread from a focus and to elevate the Ca2+ concentration in the oocyte. Activation of endogenous muscarinic or angiotensin II receptors in noninjected oocytes evoked a similar response. The Ca2+ rise in oocytes induced by substance K was due to internal Ca2+ mobilization and was independent of external Ca2+, since it occurred in Ca2(+)-free medium fortified with 2 mM EGTA. The Ca2+ imaging was well correlated with ion current measurements of voltage-clamped oocytes. Imaging, in addition to detecting the spatial spread of Ca2+ across the cell, was at least as sensitive as voltage clamping and much faster when screening oocytes for the expression of receptor mRNAs that stimulate Ca2+ mobilization. While it is known that fertilization of Xenopus eggs causes a spreading wave of Ca2+ mobilization, we found that activation of either native or newly expressed receptors in oocytes causes a similar change in Ca2+ distribution.
Brooker
G
G
Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, Washington, DC 20007.
Seki
T
T
Croll
D
D
Wahlestedt
C
C
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004594:Electrophysiology / Q000379:methods; D005260:Female; D007700:Kinetics; D008845:Microinjections; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D012333:RNA, Messenger / Q000008:administration & dosage / Q000235:genetics; D018041:Receptors, Neurokinin-2; D017981:Receptors, Neurotransmitter / Q000235:genetics / Q000502:physiology*; D013997:Time Factors; D014982:Xenopus laevis
D012333:RNA, Messenger; D018041:Receptors, Neurokinin-2; D017981:Receptors, Neurotransmitter; D004533:Egtazic Acid; D002217:Carbachol; D002118:Calcium