title: Unexpected structural and functional consequences of the R33Q homozygous mutation in cardiac calsequestrin: a complex arrhythmogenic cascade in a knock in mouse model.
abstract: Catecholaminergic polymorphic ventricular tachycardia (CPVT) is an inherited arrhythmogenic disorder characterized by life threatening arrhythmias elicited by physical and emotional stress in young individuals. The recessive form of CPVT is associated with mutation in the cardiac calsequestrin gene (CASQ2). We engineered and characterized a homozygous CASQ2(R33Q/R33Q) mouse model that closely mimics the clinical phenotype of CPVT patients. CASQ2(R33Q/R33Q) mice develop bidirectional VT on exposure to environmental stress whereas CASQ2(R33Q/R33Q) myocytes show reduction of the sarcoplasmic reticulum (SR) calcium content, adrenergically mediated delayed (DADs) and early (EADs) afterdepolarizations leading to triggered activity. Furthermore triadin, junctin, and CASQ2-R33Q proteins are significantly decreased in knock-in mice despite normal levels of mRNA, whereas the ryanodine receptor (RyR2), calreticulin, phospholamban, and SERCA2a-ATPase are not changed. Trypsin digestion studies show increased susceptibility to proteolysis of mutant CASQ2. Despite normal histology, CASQ2(R33Q/R33Q) hearts display ultrastructural changes such as disarray of junctional electron-dense material, referable to CASQ2 polymers, dilatation of junctional SR, yet normal total SR volume. Based on the foregoings, we propose that the phenotype of the CASQ2(R33Q/R33Q) CPVT mouse model is portrayed by an unexpected set of abnormalities including (1) reduced CASQ2 content, possibly attributable to increased degradation of CASQ2-R33Q, (2) reduction of SR calcium content, (3) dilatation of junctional SR, and (4) impaired clustering of mutant CASQ2.
authors.affiliation: Molecular Cardiology, IRCCS Maugeri Foundation, Pavia, Italy.
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Rizzi
authors.forename: Nian
authors.initials: N
authors.lastname: Liu
authors.forename: Carlo
authors.initials: C
authors.lastname: Napolitano
authors.forename: Alessandra
authors.initials: A
authors.lastname: Nori
authors.forename: Federica
authors.initials: F
authors.lastname: Turcato
authors.forename: Barbara
authors.initials: B
authors.lastname: Colombi
authors.forename: Silvio
authors.initials: S
authors.lastname: Bicciato
authors.forename: Diego
authors.initials: D
authors.lastname: Arcelli
authors.forename: Alessandro
authors.initials: A
authors.lastname: Spedito
authors.forename: Mario
authors.initials: M
authors.lastname: Scelsi
authors.forename: Laura
authors.initials: L
authors.lastname: Villani
authors.forename: Giovanni
authors.initials: G
authors.lastname: Esposito
authors.forename: Simona
authors.initials: S
authors.lastname: Boncompagni
authors.forename: Feliciano
authors.initials: F
authors.lastname: Protasi
authors.forename: Pompeo
authors.initials: P
authors.lastname: Volpe
authors.forename: Silvia G
authors.initials: SG
authors.lastname: Priori
mesh_terms: D000818:Animals; D001145:Arrhythmias, Cardiac / Q000209:etiology / Q000235:genetics*; D002118:Calcium / Q000032:analysis; D002155:Calsequestrin / Q000235:genetics* / Q000502:physiology; D004195:Disease Models, Animal; D004594:Electrophysiology; D006720:Homozygote; D051379:Mice; D008822:Mice, Transgenic; D020125:Mutation, Missense / Q000502:physiology*; D010641:Phenotype; D012519:Sarcoplasmic Reticulum / Q000737:chemistry
chemical_list: D002155:Calsequestrin; C514388:casq2 protein, mouse; D002118:Calcium