title: Knock-in mice harboring a Ca(2+) desensitizing mutation in cardiac troponin C develop early onset dilated cardiomyopathy.
abstract: The physiological consequences of aberrant Ca(2+) binding and exchange with cardiac myofilaments are not clearly understood. In order to examine the effect of decreasing Ca(2+) sensitivity of cTnC on cardiac function, we generated knock-in mice carrying a D73N mutation (not known to be associated with heart disease in human patients) in cTnC. The D73N mutation was engineered into the regulatory N-domain of cTnC in order to reduce Ca(2+) sensitivity of reconstituted thin filaments by increasing the rate of Ca(2+) dissociation. In addition, the D73N mutation drastically blunted the extent of Ca(2+) desensitization of reconstituted thin filaments induced by cTnI pseudo-phosphorylation. Compared to wild-type mice, heterozygous knock-in mice carrying the D73N mutation exhibited a substantially decreased Ca(2+) sensitivity of force development in skinned ventricular trabeculae. Kaplan-Meier survival analysis revealed that median survival time for knock-in mice was 12 weeks. Echocardiographic analysis revealed that knock-in mice exhibited increased left ventricular dimensions with thinner walls. Echocardiographic analysis also revealed that measures of systolic function, such as ejection fraction (EF) and fractional shortening (FS), were dramatically reduced in knock-in mice. In addition, knock-in mice displayed electrophysiological abnormalities, namely prolonged QRS and QT intervals. Furthermore, ventricular myocytes isolated from knock-in mice did not respond to β-adrenergic stimulation. Thus, knock-in mice developed pathological features similar to those observed in human patients with dilated cardiomyopathy (DCM). In conclusion, our results suggest that decreasing Ca(2+) sensitivity of the regulatory N-domain of cTnC is sufficient to trigger the development of DCM.
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, University of Houston Houston, TX, USA.
authors.forename: Bradley K
authors.initials: BK
authors.lastname: McConnell
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, University of Houston Houston, TX, USA.
authors.forename: Sonal
authors.initials: S
authors.lastname: Singh
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, University of Houston Houston, TX, USA.
authors.forename: Qiying
authors.initials: Q
authors.lastname: Fan
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, University of Houston Houston, TX, USA.
authors.forename: Adriana
authors.initials: A
authors.lastname: Hernandez
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, University of Houston Houston, TX, USA.
authors.forename: Jesus P
authors.initials: JP
authors.lastname: Portillo
authors.affiliation: Division of Biosciences, College of Dentistry, The Ohio State University Columbus, OH, USA.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Reiser
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, University of Houston Houston, TX, USA.
authors.forename: Svetlana B
authors.initials: SB
authors.lastname: Tikunova
keywords: calcium; cardiac muscle; dilated cardiomyopathy; knock-in mouse; troponin C