Measurement of the dynamics of exocytosis and vesicle retrieval at cell populations using a quartz crystal microbalance.
The quartz crystal microbalance-dissipation technique (QCM-D) is used in two different measurement strategies to monitor the mass change and rigidity of populations of excitable cells during exocytosis and subsequent retrieval of dense-core vesicles. Two cell lines, NG 108-15 and PC 12, were grown to confluence on piezoelectric quartz crystals and were examined separately to demonstrate differences in release and retrieval with cells of different morphology, size, and number of dense-core vesicles. Stimulating the cells to exocytosis with media containing an elevated potassium concentration resulted in an increase in the frequency response corresponding to loss of mass from the cells owing to release of vesicles. In Ca2+-free media, the response was completely abolished. The amplitude and peak area in the frequency response corresponding to mass change with stimulated release was larger for PC 12 cells than for NG 108-15 cells, whereas the initial rate constants for the frequency responses were similar. The data suggest (1) that a greater number and larger size of vesicles in PC 12 cells results in a greater amount of release from these cells vs NG 108-15 cells, (2) the recycling of vesicles utilizes similar fusion/retrieval mechanisms in both cell types, (3) that the control of excess retrieval might be related to the number and size of released vesicles, and (4) that measured retrieval has a rapid onset, masking exocytosis and implying a rapid retrieval mechanism in the early stages of release. These results demonstrate that measurements of complex dynamic processes relating to dense-core vesicle release and retrieval can be simultaneously accomplished using the QCM-D technique.
Cans
A S
AS
Department of Chemistry, Göteborg University, Sweden.
Höök
F
F
Shupliakov
O
O
Ewing
A G
AG
Eriksson
P S
PS
Brodin
L
L
Orwar
O
O
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D005089:Exocytosis / Q000502:physiology*; D051379:Mice; D008854:Microscopy, Electron; D036103:Nanotechnology / Q000295:instrumentation; D051381:Rats