Neonatal Rat Cardiomyocytes - A Model for the Study of Morphological, Biochemical and Electrophysiological Characteristics of the Heart

Chlopčíková, Šárka; Psotová, Jitka; Miketová, Petra

doi:10.5507/bp.2001.011

Cited by 106 publications

(83 citation statements)

References 67 publications

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“…Furthermore, studies have demonstrated that cultured neonatal cardiomyocytes form functional electromechanical coupling more comparable with an in situ heart (48). Adult rat cardiomyocytes are particularly sensitive to culture conditions; they lack resistance to cellular disintegration and quickly lose their contraction capability (6).…”

Section: Discussionmentioning

confidence: 99%

Ultrasound-induced modulation of cardiac rhythm in neonatal rat ventricular cardiomyocytes

Fleischman¹,

Vecchio

Sunny

et al. 2015

Journal of Applied Physiology

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Fleischman A, Vecchio C, Sunny Y, Bawiec CR, Lewin PA, Kresh JY, Kohut AR. Ultrasound-induced modulation of cardiac rhythm in neonatal rat ventricular cardiomyocytes. J Appl Physiol 118: 1423-1428, 2015. First published April 9, 2015 doi:10.1152/japplphysiol.00980.2014.-Isolated neonatal rat ventricular cardiomyocytes were used to study the influence of ultrasound on the chronotropic response in a tissue culture model. The beat frequency of the cells, varying from 40 to 90 beats/min, was measured based upon the translocation of the nuclear membrane captured by a high-speed camera. Ultrasound pulses (frequency ϭ 2.5 MHz) were delivered at 300-ms intervals [3.33 Hz pulse repetition frequency (PRF)], in turn corresponding to 200 pulses/min. The intensity of acoustic energy and pulse duration were made variable, 0.02-0.87 W/cm 2 and 1-5 ms, respectively. In 57 of 99 trials, there was a noted average increase in beat frequency of 25% with 8-s exposures to ultrasonic pulses. Applied ultrasound energy with a spatial peak time average acoustic intensity (Ispta) of 0.02 W/cm 2 and pulse duration of 1 ms effectively increased the contraction rate of cardiomyocytes (P Ͻ 0.05). Of the acoustic power tested, the lowest level of acoustic intensity and shortest pulse duration proved most effective at increasing the electrophysiological responsiveness and beat frequency of cardiomyocytes. Determining the optimal conditions for delivery of ultrasound will be essential to developing new models for understanding mechanoelectrical coupling (MEC) and understanding novel nonelectrical pacing modalities for clinical applications.

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Section: Discussionmentioning

confidence: 99%

Ultrasound-induced modulation of cardiac rhythm in neonatal rat ventricular cardiomyocytes

Fleischman¹,

Vecchio

Sunny

et al. 2015

Journal of Applied Physiology

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“…Cardiomyocytes were centrifuged, washed, and then preplated (3 times, 1 h each to remove nonmuscle cells and cell debris) in growth medium (see below). The isolation procedure has been previously described (11). Untransfected neonatal mouse cardiomyocytes showed spontaneous beating within 2 days after the isolation procedure, and this could be observed for up to 10 days of cell culture.…”

Section: Methodsmentioning

confidence: 99%

Properties of WT and mutant hERG K⁺ channels expressed in neonatal mouse cardiomyocytes

Lin

Holzem

Anson

et al. 2010

American Journal of Physiology-Heart and Circulatory Physiology

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Mutations in humanether-a-go-go-related gene 1 (hERG) are linked to long QT syndrome type 2 (LQT2). hERG encodes the pore-forming ␣-subunits that coassemble to form rapidly activating delayed rectifier K ϩ current in the heart. LQT2-linked missense mutations have been extensively studied in noncardiac heterologous expression systems, where biogenic (protein trafficking) and biophysical (gating and permeation) abnormalities have been postulated to underlie the loss-of-function phenotype associated with LQT2 channels. Little is known about the properties of LQT2-linked hERG channel proteins in native cardiomyocyte systems. In this study, we expressed wild-type (WT) hERG and three LQT2-linked mutations in neonatal mouse cardiomyocytes and studied their electrophysiological and biochemical properties. Compared with WT hERG channels, the LQT2 missense mutations G601S and N470D hERG exhibited altered protein trafficking and underwent pharmacological correction, and N470D hERG channels gated at more negative voltages. The ⌬Y475 hERG deletion mutation trafficked similar to WT hERG channels, gated at more negative voltages, and had rapid deactivation kinetics, and these properties were confirmed in both neonatal mouse cardiomyocyte and human embryonic kidney (HEK)-293 cell expression systems. Differences between the cardiomyocytes and HEK-293 cell expression systems were that hERG current densities were reduced 10-fold and deactivation kinetics were accelerated 1.5-to 2-fold in neonatal mouse cardiomyocytes. An important finding of this work is that pharmacological correction of trafficking-deficient LQT2 mutations, as a potential innovative approach to therapy, is possible in native cardiac tissue.

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“…17 More than 90% of cells were cardiomyocytes (positive for ␣-actinin). Adenovirus-mediated transfection efficiency in cardiomyocytes is 90% to 95%.…”

Section: Rat Neonatal Cardiomyocytesmentioning

confidence: 99%

Functional Role of Phosphodiesterase 3 in Cardiomyocyte Apoptosis

et al. 2005

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Background-Myocyte apoptosis plays an important role in pathological cardiac remodeling and the progression of heart failure. cAMP signaling is crucial in the regulation of myocyte apoptosis and cardiac remodeling. Multiple cAMP-hydrolyzing phosphodiesterases (PDEs), such as PDE3 and PDE4, coexist in cardiomyocytes and elicit differential temporal/spatial regulation of cAMP signaling. However, the role of PDE3 and PDE4 in the regulation of cardiomyocyte apoptosis remains unclear. Although chronic treatment with PDE3 inhibitors increases mortality in patients with heart failure, the contribution of PDE3 expression/activity in heart failure is not well known. Methods and Results-In this study we report that PDE3A expression and activity were significantly reduced in human failing hearts as well as mouse hearts with chronic pressure overload. In primary cultured cardiomyocytes, chronic inhibition of PDE3 but not PDE4 activity by pharmacological agents or adenovirus-delivered antisense PDE3A promoted cardiomyocyte apoptosis. Both angiotensin II (Ang II) and the ␤-adrenergic receptor agonist isoproterenol selectively induced a sustained downregulation of PDE3A expression and induced cardiomyocyte apoptosis. Restoring PDE3A via adenovirus-delivered expression of wild-type PDE3A1 completely blocked Ang II-and isoproterenolinduced cardiomyocyte apoptosis, suggesting the critical role of PDE3A reduction in cardiomyocyte apoptosis. Moreover, we defined a crucial role for inducible cAMP early repressor expression in PDE3A reduction-mediated cardiomyocyte apoptosis. Conclusions-Our results suggest that PDE3A reduction and consequent inducible cAMP early repressor induction are critical events in Ang II-and isoproterenol-induced cardiomyocyte apoptosis and may contribute to the development of heart failure. Drugs that maintain PDE3A function may represent an attractive therapeutic approach to treat heart failure.

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Neonatal Rat Cardiomyocytes - A Model for the Study of Morphological, Biochemical and Electrophysiological Characteristics of the Heart

Cited by 106 publications

References 67 publications

Ultrasound-induced modulation of cardiac rhythm in neonatal rat ventricular cardiomyocytes

Ultrasound-induced modulation of cardiac rhythm in neonatal rat ventricular cardiomyocytes

Properties of WT and mutant hERG K⁺ channels expressed in neonatal mouse cardiomyocytes

Functional Role of Phosphodiesterase 3 in Cardiomyocyte Apoptosis

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Neonatal Rat Cardiomyocytes - A Model for the Study of Morphological, Biochemical and Electrophysiological Characteristics of the Heart

Cited by 106 publications

References 67 publications

Ultrasound-induced modulation of cardiac rhythm in neonatal rat ventricular cardiomyocytes

Ultrasound-induced modulation of cardiac rhythm in neonatal rat ventricular cardiomyocytes

Properties of WT and mutant hERG K+ channels expressed in neonatal mouse cardiomyocytes

Functional Role of Phosphodiesterase 3 in Cardiomyocyte Apoptosis

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Properties of WT and mutant hERG K⁺ channels expressed in neonatal mouse cardiomyocytes