title: Responses of neurons to extreme osmomechanical stress.
abstract: Neurons are often regarded as fragile cells, easily destroyed by mechanical and osmotic insult. The results presented here demonstrate that this perception needs revision. Using extreme osmotic swelling, we show that molluscan neurons are astonishingly robust. In distilled water, a heterogeneous population of Lymnaea stagnalis CNS neurons swelled to several times their initial volume, yet had a ST50 (survival time for 50% of cells) > 60 min. Cells that were initially bigger survived longer. On return to normal medium, survivors were able, over the next 24 hr, to rearborize. Reversible membrane capacitance changes corresponding to about 0.7 muF/cm2 of apparent surface area accompanied neuronal swelling and shrinking in hypo- and hyperosmotic solutions; reversible changes in cell surface area evidently contributed to the neurons' ability to accommodate hydrostatic pressures then recover. The reversible membrane area/capacitance changes were not dependent on extracellular Ca2+. Neurons were monitored for potassium currents during direct mechanical inflation and during osmotically driven inflation. The latter but not the former stimulus routinely elicited small potassium currents, suggesting that tension increases activate the currents only if additional disruption of the cortex has occurred. Under stress in distilled water, a third of the neurons displayed a quite unexpected behavior: prolonged writhing of peripheral regions of the soma. This suggested that a plasma membrane-linked contractile machinery (presumably actomyosin) might contribute to the neurons' mechano-osmotic robustness by restricting water influx. Consistent with this possibility, 1 mM N-ethyl-maleimide, which inhibits myosin ATPase, decreased the ST50 to 18 min, rendered the survival time independent of initial size, and abolished writhing activity. For neurons, active mechanical resistance of the submembranous cortex, along with the mechanical compliance supplied by insertion or eversion of membrane stores may account for the ability to withstand diverse mechanical stresses. Mechanical robustness such as that displayed here could be an asset during neuronal outgrowth or regeneration.
authors.affiliation: Loeb Institute, Ottawa Civic Hospital, Ontario, Canada.
authors.forename: X
authors.initials: X
authors.lastname: Wan
authors.forename: J A
authors.initials: JA
authors.lastname: Harris
authors.forename: C E
authors.initials: CE
authors.lastname: Morris
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000502:physiology*; D048429:Cell Size; D002470:Cell Survival; D002478:Cells, Cultured; D004594:Electrophysiology; D008195:Lymnaea / Q000502:physiology; D008564:Membrane Potentials; D009474:Neurons / Q000502:physiology*; D009994:Osmolar Concentration; D013314:Stress, Mechanical
chemical_list: D002118:Calcium