Astrocyte swelling leads to membrane unfolding, not membrane insertion.
The mechanisms mediating the release of chemical transmitters from astrocytes are the subject of intense research. Recent experiments have shown that hypotonic conditions stimulate the release of glutamate and ATP from astrocytes, but a mechanistic understanding of this process is not available. To determine whether hypotonicity activates the process of regulated exocytosis, we monitored membrane capacitance by the whole-cell patch-clamp technique whilst a hypotonic medium was applied to cultured astrocytes. If exocytosis is triggered under hypotonic conditions, as it is following increases in cytosolic calcium, a net increase in membrane surface area, monitored by measuring the whole-cell membrane capacitance, is expected. Simultaneous measurements of cell size and whole-cell membrane conductance and surface area demonstrated that hypotonic medium (210 mOsm for 200 s) resulted in an increase in membrane conductance and in the swelling of cultured astrocytes by an average of 40%, as monitored by cell cross-sectional area, but without any corresponding change in membrane surface area. As we have demonstrated that capacitance measurements have the sensitivity to detect increases in cell surface area as small as 0.5%, we conclude that cell swelling occurs via an exocytosis-independent mechanism, probably involving the unfolding of the plasma membrane.
Pangrsic
Tina
T
Laboratory of Neuroendocrinology-Molecular Cell Physiology, Institute of Pathophysiology, Medical Faculty, University of Ljubljana, Ljubljana, Slovenia.
Potokar
Maja
M
Haydon
Philip G
PG
Zorec
Robert
R
Kreft
Marko
M
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D025802:Electric Capacitance; D005089:Exocytosis / Q000502:physiology*; D018698:Glutamic Acid / Q000378:metabolism; D007038:Hypotonic Solutions / Q000494:pharmacology; D008564:Membrane Potentials / Q000502:physiology; D009997:Osmotic Pressure; D018408:Patch-Clamp Techniques; D051381:Rats; D014882:Water-Electrolyte Balance / Q000502:physiology*
D007038:Hypotonic Solutions; D018698:Glutamic Acid; D000255:Adenosine Triphosphate; D002118:Calcium