SERCA Cys674 sulphonylation and inhibition of L-type Ca2+ influx contribute to cardiac dysfunction in endotoxemic mice, independent of cGMP synthesis.
The goal of this study was to identify the cellular mechanisms responsible for cardiac dysfunction in endotoxemic mice. We aimed to differentiate the roles of cGMP [produced by soluble guanylyl cyclase (sGC)] versus oxidative posttranslational modifications of Ca(2+) transporters. C57BL/6 mice [wild-type (WT) mice] were administered lipopolysaccharide (LPS; 25 μg/g ip) and euthanized 12 h later. Cardiomyocyte sarcomere shortening and Ca(2+) transients (ΔCai) were depressed in LPS-challenged mice versus baseline. The time constant of Ca(2+) decay (τCa) was prolonged, and sarcoplasmic reticulum Ca(2+) load (CaSR) was depressed in LPS-challenged mice (vs. baseline), indicating decreased activity of sarco(endo)plasmic Ca(2+)-ATPase (SERCA). L-type Ca(2+) channel current (ICa,L) was also decreased after LPS challenge, whereas Na(+)/Ca(2+) exchange activity, ryanodine receptors leak flux, or myofilament sensitivity for Ca(2+) were unchanged. All Ca(2+)-handling abnormalities induced by LPS (the decrease in sarcomere shortening, ΔCai, CaSR, ICa,L, and τCa prolongation) were more pronounced in mice deficient in the sGC main isoform (sGCα1(-/-) mice) versus WT mice. LPS did not alter the protein expression of SERCA and phospholamban in either genotype. After LPS, phospholamban phosphorylation at Ser(16) and Thr(17) was unchanged in WT mice and was increased in sGCα1(-/-) mice. LPS caused sulphonylation of SERCA Cys(674) (as measured immunohistochemically and supported by iodoacetamide labeling), which was greater in sGCα1(-/-) versus WT mice. Taken together, these results suggest that cardiac Ca(2+) dysregulation in endotoxemic mice is mediated by a decrease in L-type Ca(2+) channel function and oxidative posttranslational modifications of SERCA Cys(674), with the latter (at least) being opposed by sGC-released cGMP.
Cardiovascular Medicine Section, Department of Medicine, Boston University Medical Center, Boston, Massachusetts;
Ion A
IA
Hobai
Emmanuel S
ES
Buys
Justin C
JC
Morse
Jessica
J
Edgecomb
Eric H
EH
Weiss
Antonis A
AA
Armoundas
Xiuyun
X
Hou
Alok R
AR
Khandelwal
Deborah A
DA
Siwik
Peter
P
Brouckaert
Richard A
RA
Cohen
Wilson S
WS
Colucci
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020746:Calcium Channels, L-Type / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D006152:Cyclic GMP / Q000096:biosynthesis; D003545:Cysteine / Q000378:metabolism; D019446:Endotoxemia / Q000378:metabolism*; D006162:Guanylate Cyclase / Q000235:genetics; D006321:Heart / Q000503:physiopathology*; D008070:Lipopolysaccharides; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009206:Myocardium / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism; D012518:Sarcomeres; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism*; D019831:Sodium-Calcium Exchanger / Q000378:metabolism; D000099211:Phospholamban
D002118:Calcium; D020746:Calcium Channels, L-Type; D002135:Calcium-Binding Proteins; D006152:Cyclic GMP; D003545:Cysteine; D006162:Guanylate Cyclase; D008070:Lipopolysaccharides; D019837:Ryanodine Receptor Calcium Release Channel; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D019831:Sodium-Calcium Exchanger; D000099211:Phospholamban
cardiomyopathy; contractility; cysteine; oxidative stress; sarco(endo)plasmic reticulum Ca2+-ATPase; sarcoplasmic reticulum; sepsis