Simultaneous detection of catecholamine exocytosis and Ca2+ release from single bovine chromaffin cells using a dual microsensor.
A dual microsensor with a 5 microns radius was fabricated to detect simultaneously Ca2+ and catecholamines following their secretion from individual biological cells. Detection of Ca2+ was based on changes in fluorescence as a result of its binding with a surface-attached dye, and catecholamines were detected by amperometry. The fluorescent dye employed, calcium green-1 dextran, is a selective chelator for Ca2+. It was attached to the tip of a carbon fiber electrode by cross-linking with 5% glutaraldehyde. The dual microsensor has a subsecond response time for both Ca2+ and catecholamine concentration changes. Ca2+ concentrations of 100 nM can be detected, while the detection limit for catecholamine is in the micromolar range. The utility of the dual microsensor was evaluated at the surface of bovine adrenal medullary cells. Release of catecholamines by exocytosis was evoked by transient application of histamine. This was detected by amperometry, and it was found to be accompanied by Ca2+ release, as measured by fluorescence from the same sensor.
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Department of Chemistry, University of North Carolina, Chapel Hill 27599-3290, USA.
Wightman
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RM
D000313:Adrenal Medulla / Q000166:cytology / Q000378:metabolism; D000818:Animals; D015374:Biosensing Techniques; D002118:Calcium / Q000378:metabolism*; D002395:Catecholamines / Q000032:analysis* / Q000378:metabolism; D002417:Cattle; D019439:Chromaffin Cells / Q000378:metabolism*; D005089:Exocytosis / Q000502:physiology*; D005456:Fluorescent Dyes
D002395:Catecholamines; D005456:Fluorescent Dyes; D002118:Calcium