title: DHPR activation underlies SR Ca2+ release induced by osmotic stress in isolated rat skeletal muscle fibers.
abstract: Changes in skeletal muscle volume induce localized sarcoplasmic reticulum (SR) Ca(2+) release (LCR) events, which are sustained for many minutes, suggesting a possible signaling role in plasticity or pathology. However, the mechanism by which cell volume influences SR Ca(2+) release is uncertain. In the present study, rat flexor digitorum brevis fibers were superfused with isoosmotic Tyrode's solution before exposure to either hyperosmotic (404 mOsm) or hypoosmotic (254 mOsm) solutions, and the effects on cell volume, membrane potential (E(m)), and intracellular Ca(2+) ([Ca(2+)](i)) were determined. To allow comparison with previous studies, solutions were made hyperosmotic by the addition of sugars or divalent cations, or they were made hypoosmotic by reducing [NaCl](o). All hyperosmotic solutions induced a sustained decrease in cell volume, which was accompanied by membrane depolarization (by 14-18 mV; n = 40) and SR Ca(2+) release. However, sugar solutions caused a global increase in [Ca(2+)](i), whereas solutions made hyperosmotic by the addition of divalent cations only induced LCR. Decreasing osmolarity induced an increase in cell volume and a negative shift in E(m) (by 15.04 +/- 1.85 mV; n = 8), whereas [Ca(2+)](i) was unaffected. However, on return to the isoosmotic solution, restoration of cell volume and E(m) was associated with LCR. Both global and localized SR Ca(2+) release were abolished by the dihydropyridine receptor inhibitor nifedipine by sustained depolarization of the sarcolemmal or by the addition of the ryanodine receptor 1 inhibitor tetracaine. Inhibitors of the Na-K-2Cl (NKCC) cotransporter markedly inhibited the depolarization associated with hyperosmotic shrinkage and the associated SR Ca(2+) release. These findings suggest (1) that the depolarization that accompanies a decrease in cell volume is the primary event leading to SR Ca(2+) release, and (2) that volume-dependent regulation of the NKCC cotransporter contributes to the observed changes in E(m). The differing effects of the osmotic agents can be explained by the screening of fixed charges by divalent ions.
authors.affiliation: Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, England, UK.
authors.forename: James D
authors.initials: JD
authors.lastname: Pickering
authors.forename: Ed
authors.initials: E
authors.lastname: White
authors.forename: Adrian M
authors.initials: AM
authors.lastname: Duke
authors.forename: Derek S
authors.initials: DS
authors.lastname: Steele
mesh_terms: D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000187:drug effects / Q000378:metabolism*; D020013:Calcium Signaling; D048429:Cell Size; D002712:Chlorides / Q000378:metabolism; D066298:In Vitro Techniques; D007552:Isotonic Solutions; D007700:Kinetics; D008297:Male; D008564:Membrane Potentials; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000378:metabolism*; D009543:Nifedipine / Q000494:pharmacology; D009997:Osmotic Pressure; D051381:Rats; D017208:Rats, Wistar; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects / Q000378:metabolism; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism*; D028021:Sodium-Potassium-Chloride Symporters / Q000378:metabolism; D013748:Tetracaine / Q000494:pharmacology
chemical_list: D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D002712:Chlorides; D007552:Isotonic Solutions; D019837:Ryanodine Receptor Calcium Release Channel; D028021:Sodium-Potassium-Chloride Symporters; C037692:Tyrode's solution; D013748:Tetracaine; D009543:Nifedipine