title: Triadins modulate intracellular Ca(2+) homeostasis but are not essential for excitation-contraction coupling in skeletal muscle.
abstract: 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.
authors.affiliation: Department of Anesthesiology, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Shen
authors.forename: Clara
authors.initials: C
authors.lastname: Franzini-Armstrong
authors.forename: Jose R
authors.initials: JR
authors.lastname: Lopez
authors.forename: Larry R
authors.initials: LR
authors.lastname: Jones
authors.forename: Yvonne M
authors.initials: YM
authors.lastname: Kobayashi
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: W Glenn L
authors.initials: WG
authors.lastname: Kerrick
authors.forename: Anthony H
authors.initials: AH
authors.lastname: Caswell
authors.forename: James D
authors.initials: JD
authors.lastname: Potter
authors.forename: Todd
authors.initials: T
authors.lastname: Miller
authors.forename: Paul D
authors.initials: PD
authors.lastname: Allen
authors.forename: Claudio F
authors.initials: CF
authors.lastname: Perez
mesh_terms: 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
chemical_list: 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