Monitoring the effect of osmotic stress on secretory vesicles and exocytosis
Artikel i vetenskaplig tidskrift, 2018

Amperometry recording of cells subjected to osmotic shock show that secretory cells respond to this physical stress by reducing the exocytosis activity and the amount of neurotransmitter released from vesicles in single exocytosis events. It has been suggested that the reduction in neurotransmitters expelled is due to alterations in membrane biophysical properties when cells shrink in response to osmotic stress and with assumptions made that secretory vesicles in the cell cytoplasm are not affected by extracellular osmotic stress. Amperometry recording of exocytosis monitors what is released from cells the moment a vesicle fuses with the plasma membrane, but does not provide information on the vesicle content before the vesicle fusion is triggered. Therefore, by combining amperometry recording with other complementary analytical methods that are capable of characterizing the secretory vesicles before exocytosis at cells is triggered offers a broader overview for examining how secretory vesicles and the exocytosis process are affected by osmotic shock. We here describe how complementing amperometry recording with intracellular electrochemical cytometry and transmission electron microscopy (TEM) imaging can be used to characterize alterations in secretory vesicles size and neurotransmitter content at chromaffin cells in relation to exocytosis activity before and after exposure to osmotic stress. By linking the quantitative information gained from experiments using all three analytical methods, conclusions were previously made that secretory vesicles respond to extracellular osmotic stress by shrinking in size and reducing the vesicle quantal size to maintain a constant vesicle neurotransmitter concentration. Hence, this gives some clarification regarding why vesicles, in response to osmotic stress, reduce the amount neurotransmitters released during exocytosis release. The amperometric recordings here indicate this is a reversible process and that vesicle after osmotic shock are refilled with neurotransmitters when placed cells are reverted into an isotonic environment.

Osmotic pressure

Quantal size

Transmission electron microscopy

Amperometry

Neurotransmitter concentration

Intracellular cytometry

Exocytosis

Dense core vesicles

Chromaffin cells

Neuroscience

Författare

Hoda Mashadi Fathali

Chalmers, Kemi och kemiteknik, Kemi och biokemi, Analytisk kemi

Johan Dunevall

Chalmers, Kemi och kemiteknik, Kemi och biokemi, Fysikalisk kemi

Soodabeh Majdi

Göteborgs universitet

Ann-Sofie Cans

Chalmers, Kemi och kemiteknik, Kemi och biokemi

Journal of Visualized Experiments

1940-087X (ISSN)

Vol. 2018 e56537

Ämneskategorier

Cellbiologi

Neurovetenskaper

Cell- och molekylärbiologi