Cholecystokinin-octapeptide affects the fluorescence signal of a single pancreatic acinar cell loaded with the acrylodan-labelled MARCKS peptide, a protein kinase C substrate

authored by: A Ngezahayo, F Lang, H A Kolb
Abstract: We used a fluorescent derivative of the myristoylated, alanine-rich C kinase substrate (MARCKS) peptide as a probe for protein kinase C (PKC) activation by cholecystokinin-octapeptide (CCK-8) in isolated pancreatic acinar cell pairs. The diffusion of the acrylodan-labelled MARCKS-peptide into the cell interior could be monitored by the increase of fluorescence in the whole-cell patch-clamp configuration. Addition of 10 pM CCK-8 to the bath induced repetitive fluctuations of the fluorescent signal in the time range of 4-5 min. With 1 nM CCK-8 a sustained decrease of the signal was observed. Addition of polymyxin B, a specific inhibitor of PKC activation, to the pipette filling solution suppressed the CCK-8-induced change of fluorescence. The data indicate activation of PKC by CCK-8 in pancreatic acinar cells and could be compared with the previously studied CCK-8-induced gap junction uncoupling.
Organisation(s): Department of Cell Physiology and Biophysics
External Organisation(s): University of Tübingen
Russian Academy of Medical Sciences - Institute of Normal Physiology
Type: Article
Journal: Pflugers Archiv European Journal of Physiology
Volume: 429
Pages: 805-8
No. of pages: 4
ISSN: 0031-6768
Publication date: 04.1995
Publication status: Published
Peer reviewed: Yes
Electronic version(s): https://doi.org/10.1007/BF00374804 (Access: Restricted)

BibTeX

@article{61aa27c3b31144ba82a0147ef1711435,
title = "Cholecystokinin-octapeptide affects the fluorescence signal of a single pancreatic acinar cell loaded with the acrylodan-labelled MARCKS peptide, a protein kinase C substrate",
abstract = "We used a fluorescent derivative of the myristoylated, alanine-rich C kinase substrate (MARCKS) peptide as a probe for protein kinase C (PKC) activation by cholecystokinin-octapeptide (CCK-8) in isolated pancreatic acinar cell pairs. The diffusion of the acrylodan-labelled MARCKS-peptide into the cell interior could be monitored by the increase of fluorescence in the whole-cell patch-clamp configuration. Addition of 10 pM CCK-8 to the bath induced repetitive fluctuations of the fluorescent signal in the time range of 4-5 min. With 1 nM CCK-8 a sustained decrease of the signal was observed. Addition of polymyxin B, a specific inhibitor of PKC activation, to the pipette filling solution suppressed the CCK-8-induced change of fluorescence. The data indicate activation of PKC by CCK-8 in pancreatic acinar cells and could be compared with the previously studied CCK-8-induced gap junction uncoupling.",
keywords = "2-Naphthylamine/analogs & derivatives, Animals, Fluorescent Dyes, Intracellular Signaling Peptides and Proteins, Kinetics, Male, Membrane Proteins, Mice, Myristoylated Alanine-Rich C Kinase Substrate, Pancreas/metabolism, Patch-Clamp Techniques, Protein Kinase C/antagonists & inhibitors, Proteins/metabolism, Sincalide/pharmacology, Spectrometry, Fluorescence",
author = "A Ngezahayo and F Lang and Kolb, {H A}",
year = "1995",
month = apr,
doi = "10.1007/BF00374804",
language = "English",
volume = "429",
pages = "805--8",
journal = "Pflugers Archiv European Journal of Physiology",
issn = "0031-6768",
publisher = "Springer Verlag",
number = "6",
}