Halothane and isoflurane effects on Ca2+ fluxes of isolated myocardial sarcoplasmic reticulum.
To elucidate better the differential myocardial depressant actions of halogenated volatile anesthetics, anesthetic-induced changes in Ca2+ accumulation, release, and Ca-ATPase (Ca2+ pump) activity of isolated canine cardiac sarcoplasmic reticulum (SR) vesicles were examined. An initial crude microsomal fraction of homogenized canine ventricle was subfractionated on a discontinuous sucrose gradient after Ca2+ loading in the presence of phosphate. Junctional SR (JSR) enriched with terminal cisternae was identified by its content of an electrophoretically verified approximately 450-kDa protein, the Ca(2+)-release channel (CaRC). When the CaRC of JSR was blocked by 1 microM ruthenium red (RR), the rate of Ca2+ uptake increased 47% as measured spectrophotometrically using the Ca-sensitive dye antipyrylazo III. A second fraction was identified as primarily longitudinal SR (LSR) based on its trace content of 450-kDa protein and 11% increase of Ca2+ uptake with RR. Halothane (0.75-2.5%) or isoflurane (2.5-4%) decreased net Ca2+ accumulation rate by either LSR or JSR, and the decrease in uptake rate of JSR was only partially reversed by addition of 1 microM RR (27% increase for isoflurane, 7% increase for halothane). Both halothane and isoflurane increased JSR ATP consumption as measured by a coupled-enzyme assay. 45Ca2+ efflux from passively loaded SR vesicles was then determined to verify that the decreased net uptake rate was due to enhanced Ca2+ efflux from vesicles. Both anesthetics increased passive Ca2+ efflux from SR vesicles in which the CaRC was blocked by 10 microM RR as well as those in which Ca2+ release via the CaRC was activated by 10 microM Ca2+.(ABSTRACT TRUNCATED AT 250 WORDS)
Frazer
M J
MJ
Department of Anesthesiology, University of Virginia Health Sciences Center, Charlottesville 22908.
Lynch
C
C
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D004285:Dogs; D006221:Halothane / Q000494:pharmacology*; D006321:Heart / Q000187:drug effects*; D066298:In Vitro Techniques; D007530:Isoflurane / Q000494:pharmacology*; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism*; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*
D007530:Isoflurane; D000252:Calcium-Transporting ATPases; D002118:Calcium; D006221:Halothane