Details
| Original language | English |
|---|---|
| Pages (from-to) | 805-8 |
| Number of pages | 4 |
| Journal | Pflugers Archiv European Journal of Physiology |
| Volume | 429 |
| Issue number | 6 |
| Publication status | Published - Apr 1995 |
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
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In: Pflugers Archiv European Journal of Physiology, Vol. 429, No. 6, 04.1995, p. 805-8.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Cholecystokinin-octapeptide affects the fluorescence signal of a single pancreatic acinar cell loaded with the acrylodan-labelled MARCKS peptide, a protein kinase C substrate
AU - Ngezahayo, A
AU - Lang, F
AU - Kolb, H A
PY - 1995/4
Y1 - 1995/4
N2 - 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.
AB - 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.
KW - 2-Naphthylamine/analogs & derivatives
KW - Animals
KW - Fluorescent Dyes
KW - Intracellular Signaling Peptides and Proteins
KW - Kinetics
KW - Male
KW - Membrane Proteins
KW - Mice
KW - Myristoylated Alanine-Rich C Kinase Substrate
KW - Pancreas/metabolism
KW - Patch-Clamp Techniques
KW - Protein Kinase C/antagonists & inhibitors
KW - Proteins/metabolism
KW - Sincalide/pharmacology
KW - Spectrometry, Fluorescence
U2 - 10.1007/BF00374804
DO - 10.1007/BF00374804
M3 - Article
C2 - 7603834
VL - 429
SP - 805
EP - 808
JO - Pflugers Archiv European Journal of Physiology
JF - Pflugers Archiv European Journal of Physiology
SN - 0031-6768
IS - 6
ER -