Optical measurement of stimulus-evoked membrane dynamics in single pancreatic acinar cells.
Stimulation of pancreatic acinar cells induces the release of digestive enzymes via the exocytotic fusion of zymogen granules and activates postfusion granule membrane retrieval and receptor cycling. In the present study, changes in membrane surface area of rat single pancreatic acinar cells were monitored by cell membrane capacitance (Cm) measurements and by the membrane fluorescent dye FM1-43. When measured with the Cm method, agonist treatment evoked a graded, transient increase in acinar cell surface area averaging 3. 5%. In contrast, a 13% increase in surface area was estimated using FM1-43, corresponding to the fusion of 48 zymogen granules at a rate of 0.5 s-1. After removal of FM1-43 from the surface-accessible membrane, a residual fluorescence signal was shown by confocal microscopy to be localized in endosome-like structures and confined to the apical regions of acinar cells. The development of an optical method for monitoring the membrane turnover of single acinar cells, in combination with measurements of Cm changes, reveals coincidence of exocytotic and endocytotic activity in acinar cells after hormonal stimulation.
Giovannucci
D R
DR
Department of Physiology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Yule
D I
DI
Stuenkel
E L
EL
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002217:Carbachol / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D003594:Cytoplasmic Granules / Q000502:physiology; D003600:Cytosol / Q000378:metabolism; D005089:Exocytosis; D005456:Fluorescent Dyes; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology*; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D008561:Membrane Fusion; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D010179:Pancreas / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D051381:Rats; D017207:Rats, Sprague-Dawley
C073804:FM1 43; D005456:Fluorescent Dyes; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D002217:Carbachol; D002118:Calcium