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Cardiovascular Research 1999 41(1):200-211; doi:10.1016/S0008-6363(98)00217-X
© 1999 by European Society of Cardiology
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Copyright © 1999, European Society of Cardiology

A soluble neuronal factor alters contractile function of ventricular myocytes without effect on troponin T isoform expression

Steven H. Greena, Rebecca M. Glovera, Penny A. Krummb, Timothy R. Wintersa and Dianne L. Atkinsb,*

aDepartment of Biological Sciences, University of Iowa, Iowa City, IA 52242, USA
bDepartment of Pediatrics, University of Iowa, Iowa City, IA 52242, USA

* Corresponding author. Tel.: +1-319-356-3540; fax: +1-319-356-4693; e-mail: dianne-atkins@uiowa.edu

Objective: The purpose of this investigation was to establish a model system to facilitate identification of the sympathetic neuronal factor(s) that promotes improved contractility in neonatal cardiac myocytes. Conditioned medium from PC12 cells with sympathetic phenotype served as the source of the neuronal factor. Methods: Contraction frequency, amplitude and velocity of cultured neonatal rat cardiac myocytes were measured by online video analysis. Interventions included in vitro sympathetic innervation, exposure to PC12 conditioned medium, neurotransmitters and antagonists. Metabolic activity was assayed by 2-deoxyglucose uptake. Troponin T isoform expression was analyzed by SDS–polyacrylamide gel electrophoresis. Results: Medium conditioned by neuronal PC12 cells induced contractility changes similar to those induced by in vitro sympathetic innervation. These effects of PC12 conditioned medium and innervation were not suppressed by adrenergic or muscarinic antagonists nor reproduced by neuropeptide Y or somatostatin. Neuronal PC12 conditioned medium but not chromaffin PC12 conditioned medium, increased metabolic activity of the myocytes as detected by [3H]-2-deoxyglucose, indicating that the effect was specific to the neuronal PC12 cells. The in vitro switch of troponin T isoform expression was not altered by exposure to PC12 conditioned medium. Conclusions: Increased contractile function induced by sympathetic innervation is reproduced by PC12 conditioned medium, but neither is mediated by sympathetic or muscarinic neurotransmitters. Troponin T isoform expression is not related to the contractility changes. This model system will allow identification of the factor(s).

KEYWORDS Cardiac myocytes; Contractile function; Neuronal factors; Sympathetic neurons; PC12 cells; Troponin T; Cell culture; Rats


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