Triadins modulate intracellular Ca(2+) homeostasis but are not essential for excitation-contraction coupling in skeletal muscle.
To unmask the role of triadin in skeletal muscle we engineered pan-triadin-null mice by removing the first exon of the triadin gene. This resulted in a total lack of triadin expression in both skeletal and cardiac muscle. Triadin knockout was not embryonic or birth-lethal, and null mice presented no obvious functional phenotype. Western blot analysis of sarcoplasmic reticulum (SR) proteins in skeletal muscle showed that the absence of triadin expression was associated with down-regulation of Junctophilin-1, junctin, and calsequestrin but resulted in no obvious contractile dysfunction. Ca(2+) imaging studies in null lumbricalis muscles and myotubes showed that the lack of triadin did not prevent skeletal excitation-contraction coupling but reduced the amplitude of their Ca(2+) transients. Additionally, null myotubes and adult fibers had significantly increased myoplasmic resting free Ca(2+).[(3)H]Ryanodine binding studies of skeletal muscle SR vesicles detected no differences in Ca(2+) activation or Ca(2+) and Mg(2+) inhibition between wild-type and triadin-null animals. Subtle ultrastructural changes, evidenced by the appearance of longitudinally oriented triads and the presence of calsequestrin in the sacs of the longitudinal SR, were present in fast but not slow twitch-null muscles. Overall, our data support an indirect role for triadin in regulating myoplasmic Ca(2+) homeostasis and organizing the molecular complex of the triad but not in regulating skeletal-type excitation-contraction coupling.
Shen
Xiaohua
X
Department of Anesthesiology, Brigham and Women's Hospital, Boston, MA 02115, USA.
Franzini-Armstrong
Clara
C
Lopez
Jose R
JR
Jones
Larry R
LR
Kobayashi
Yvonne M
YM
Wang
Ying
Y
Kerrick
W Glenn L
WG
Caswell
Anthony H
AH
Potter
James D
JD
Miller
Todd
T
Allen
Paul D
PD
Perez
Claudio F
CF
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000235:genetics; D002155:Calsequestrin / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics* / Q000502:physiology*; D005091:Exons; D005786:Gene Expression Regulation; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D006899:Mixed Function Oxygenases / Q000235:genetics; D008954:Models, Biological; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D009124:Muscle Proteins / Q000235:genetics* / Q000502:physiology*; D018482:Muscle, Skeletal / Q000378:metabolism*; D011189:Potassium Chloride / Q000378:metabolism; D012519:Sarcoplasmic Reticulum / Q000378:metabolism
D002135:Calcium-Binding Proteins; D002155:Calsequestrin; D002352:Carrier Proteins; D008565:Membrane Proteins; D009124:Muscle Proteins; C412507:junctophilin; C070048:triadin; D011189:Potassium Chloride; C498841:Asph protein, mouse; D006899:Mixed Function Oxygenases; D002118:Calcium