Abstract
Examine the feasibility of whole-cell patch-clamp recordings from the cardiac ventricular slices of newborn (P3–P7) Sprague–Dawley rats to identify a better substitute for single cardiac myocytes prepared using enzymatic treatment. High resistance seals (>1 GΏ) were obtained from cardiac ventricle tissues prepared without using enzymatic treatment. Thereafter, cell-attached and whole-cell patch-clamp techniques were used on thin cardiac slices (200 μm thick) in 2009 in the Institute of Molecular Medicine of Peking University. An averaged sodium current (n = 11 cells) was recorded in the cell-attached mode, and this displayed features similar to those previously reported for isolated rat ventricular myocytes. The outward potassium current, hyperpolarization-activated cation channel or I f channel (HCN channel), and action potential were recorded in the whole-cell mode (n = 2 cells), and the identical properties were observed from the cardiac slices. The cell-attached mode is stable and reliable for recording the ion current. The resting potential for cardiac slices measured using current-clamp recording in the whole-cell mode was −50 to −70 mV. The resting potential of cardiac slices has properties similar to those of enzyme-prepared cardiomyocytes, with the exception that it is positive. We achieved whole-cell patch-clamp recordings from cardiac slices and affirmed the feasibility and values of both cell-attached and whole-cell recording modes using this technique. Nevertheless, there remain difficulties and limitations associated with the application of whole-cell patch-clamping to cardiac slices, due primarily to the existence of large amounts of connective tissue even in newborn rats.
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References
Edwards F. A.; Konnerth A.; Sakmann B.; Takahashi T. A thin slice preparation for patch-clamp recordings from neurones of the mammalian central nervous system. Pflugers Arch. 414: 600–612; 1989.
Konnerth A. Patch-clamping in slices of mammalian CNS. Trends Neurosci. 13: 321–323; 1990.
Li G. R.; Lau C. P.; Alvin S. Heterogeneity of sodium current in atrial vs epicardial ventricular myocytes of adult guinea pig hearts. J. Mol. Cell. Cardiol. 34: 1185–1194; 2002.
Moyer Jr. J. R.; Brown T. H. Patch-clamp techniques applied to brain slices. Neuromethods 26: 233–258; 1995.
Wu B.; Wang Y. M.; Xiong W.; Zheng L. H.; Fu C. L.; Bruce I. C.; Zhang C.; Zhou Z. Optimization of a multi-channel puffer system for rapid delivery of solutions during patch-clamp experiments. Front. Biosci. 10: 761–767; 2005.
Xu Z. Q.; Pieribone V. A.; Zhang X.; Grillner S.; Hokfelt T. A functional role for nitric oxide in locus coeruleus: immunohistochemical and electrophysiological studies. Exp. Brain Res. 98: 75–83; 1994.
Acknowlegements
The study was performed in the IMM of Peking University. We thank Prof. Zhuan Zhou for his directions and support, thank Hongping Huang and Bo Zhang of IMM for their kindly help for preparation of the cardiac slices.
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Editor: J. Denry Sato
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Huang, D., Li, J. The feasibility and limitation of patch-clamp recordings from neonatal rat cardiac ventricular slices. In Vitro Cell.Dev.Biol.-Animal 47, 269–272 (2011). https://doi.org/10.1007/s11626-011-9387-6
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DOI: https://doi.org/10.1007/s11626-011-9387-6