Calcium-dependent transmitter secretion reconstituted in Xenopus oocytes: requirement for synaptophysin.
Calcium-dependent glutamate secretion was reconstituted in Xenopus oocytes by injecting the oocyte with total rat cerebellar messenger RNA (mRNA). Co-injection of total mRNA with antisense oligonucleotides to synaptophysin message decreased the expression of synaptophysin in the oocyte and reduced the calcium-dependent secretion. A similar effect on secretion was observed for oocytes injected with total mRNA together with an antibody to rat synaptophysin. These results indicate that synaptophysin is necessary for transmitter secretion and that the oocyte expression system may be useful for dissecting the molecular events associated with the secretory process.
Alder
J
J
Department of Biological Sciences, Columbia University, New York, NY 10027.
Lu
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B
Valtorta
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Greengard
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Poo
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MM
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology*; D002531:Cerebellum / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D007700:Kinetics; D008099:Liver / Q000737:chemistry; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D009865:Oocytes / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D016708:Synaptophysin / Q000235:genetics / Q000502:physiology*; D014162:Transfection; D014981:Xenopus
D005971:Glutamates; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D016708:Synaptophysin; D000001:Calcimycin; D018698:Glutamic Acid; D002118:Calcium