MONITORING EXOCYTOSIS AND RELEASE FROM INDIVIDUAL MAST-CELLS BY CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED NATIVE FLUORESCENCE DETECTION

Citation
Sj. Lillard et al., MONITORING EXOCYTOSIS AND RELEASE FROM INDIVIDUAL MAST-CELLS BY CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED NATIVE FLUORESCENCE DETECTION, Analytical chemistry, 68(17), 1996, pp. 2897-2904
Citations number
35
Categorie Soggetti
Chemistry Analytical
Journal title
ISSN journal
00032700
Volume
68
Issue
17
Year of publication
1996
Pages
2897 - 2904
Database
ISI
SICI code
0003-2700(1996)68:17<2897:MEARFI>2.0.ZU;2-#
Abstract
The complex temporal evolution of on-column exocytotic release of sero tonin and proteins from individual rat peritoneal mast cells (RPMCs) w as monitored by using capillary electrophoresis. Laser-induced native fluorescence detection with 275-nm excitation was used, and a detectio n limit of 1.7 amol (S/N = 3; rms) was obtained for serotonin. A physi ological rurning buffer was used to ensure that the cell remained viab le throughout. The secretagogue was polymyxin B sulfate (Pmx). Followi ng the injection of a single mast cell into the capillary, electromigr ation of Pmx toward and past the cell induced degranulation and releas e of serotonin. The time course of release was registered in the elect ropherograms with subsecond resolution. Subsequent introduction of SDS caused the cell to lyse completely and allowed the residual serotonin to be quantified. The average amount of serotonin observed per RPMC w as 1.6 +/- 0.6 fmol; the average percentage of serotonin released was 28 +/- 14%. Events that are consistent with released serotonin from si ngle submicrometer granules (250 at each) were evident, each of which contained an average amount of 5.9 +/- 3 amol.