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Titolo:
Measurement of exocytosis by amperometry in adrenal chromaffin cells: Effects of clostridial neurotoxins and activation of protein kinase C on fusionpore kinetics
Autore:
Graham, ME; Fisher, RJ; Burgoyne, RD;
Indirizzi:
Univ Liverpool, Physiol Lab, Liverpool L69 3BX, Merseyside, England Univ Liverpool Liverpool Merseyside England L69 3BX , Merseyside, England
Titolo Testata:
BIOCHIMIE
fascicolo: 5, volume: 82, anno: 2000,
pagine: 469 - 479
SICI:
0300-9084(200005)82:5<469:MOEBAI>2.0.ZU;2-D
Fonte:
ISI
Lingua:
ENG
Soggetto:
SYNAPTIC VESICLE DOCKING; NEUROTRANSMITTER RELEASE; MEMBRANE-FUSION; PHORBOL ESTER; MEDULLARY CELLS; PC12 CELLS; BOTULINUM NEUROTOXINS; REGULATED EXOCYTOSIS; SNARE COMPLEXES; SEROTYPE-A;
Keywords:
exocytosis; membrane fusion; protein kinase C; chromaffin cell; calcium;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
54
Recensione:
Indirizzi per estratti:
Indirizzo: Burgoyne, RD Univ Liverpool, Physiol Lab, Crown St, Liverpool L69 3BX, Merseyside, England Univ Liverpool Crown St Liverpool Merseyside England L69 3BX
Citazione:
M.E. Graham et al., "Measurement of exocytosis by amperometry in adrenal chromaffin cells: Effects of clostridial neurotoxins and activation of protein kinase C on fusionpore kinetics", BIOCHIMIE, 82(5), 2000, pp. 469-479

Abstract

We have used carbon-fibre amperometry to examine the kinetics of individual secretory granule fusion/release events in bovine adrenal chromaffin cells. Transfection with plasmids encoding the light chains of botulinum neurotoxins (BoNTs) was used to investigate the effects of cleavage of syntaxin or SNAP-25 on exocytosis. Expression of BoNT/C1 or BoNT/E inhibited the extent of exocytosis that was evoked by application of digitonin/Ca2+ to permeabilise and stimulate single chromaffin cells. Following neurotoxin expression, the residual release events were no different from those of control cells in their magnitude and kinetics from analysis of the amperometric spikes. In contrast, activation of protein kinase C (PKC) resulted in a modification of the kinetics of single granule release events. Following phorbol ester treatment, the amperometric spikes showed a significant decrease in their total charge due to a decrease in their mean half-width with increases inthe rate of the initial rise and also the fall to baseline of the spikes. These changes were prevented by pre-treatment with the PKC inhibitor bisindolylmaleimide. These results suggest that PKC regulates the rate of fusion pore expansion and also subsequent pore closure or granule retrieval. A PKC-mediated regulation of kiss-and-run fusion may, therefore, control the extent of catecholamine release from single secretory granules. The experimental approach used here may provide further information on the protein constituents and regulation of the fusion pore machinery. (C) 2000 Societe francaise de biochimie et biologie moleculaire / Editions scientifiques et medicales Elsevier SAS.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/07/20 alle ore 22:41:12