Isolated Cardiac Muscle Assays

Terry Kenakin1

1 Glaxo Wellcome Research And Development, Research Triangle Park, North Carolina
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 4.3
DOI:  10.1002/0471141755.ph0403s00
Online Posting Date:  May, 2001
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Isolated cardiac muscle is one of the major tissues used in pharmacological research. It is compact, easily prepared, and functions as a syncytium yielding rapid responses to a variety of agents. Described in this unit are methods of preparing and handling isolated cardiac muscle, a preparation rich in autonomic receptors.

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Table of Contents

  • Reagents and Solutions
  • Commentary
  • Figures
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Basic Protocol 1:

  • Animal suitable for receptor of interest (see appropriate unit of this chapter)
  • 1× physiological salt solution (PSS) appropriate for the species in question (see recipe), 4°C
  • Carbogen gas (95% O 2/5% CO 2; tank of compressed gas with regulator capable of releasing gas under extremely low pressure, i.e., 3 to 4 lb/in2)
  • Catecholamine solution (see recipe)
  • Petri dish equipped with fine capillary tubing made of nonreactive plastic (e.g., Tygon) through which carbogen is bubbled through the solution
  • Surgical instruments: fine scissors, forceps
  • 5‐O gauge fine silk thread
  • Gold jeweler's chains or stainless steel wire hooks (optional; required when affixing muscle to recording apparatus)
  • Isometric Plexiglas tissue holder fitted for fine adjustment of resting tension, with two punctate platinum electrodes (4 to 6 mm apart) milled flat on the surface of the electrode or one punctate electrode and an external electrode (see Fig. )
  • Heated circulating water bath, set at 31° to 34°C with thermostat ±0.1°C
  • Isolated organ bath
  • Isometric transducer (Grass Instruments FT.03)
  • Electrical stimulator capable of delivering a square wave electrical stimulus, or potentiometric rate meter to measure heart rate (for spontaneously beating preparations)
  • Physiological chart recorder
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Literature Cited

Literature Cited
   Blinks, J.R. 1965. Convenient apparatus for recording contractions of isolated heart muscle. J. Appl. Physiol. 20:755‐757.
   Blinks, J.R. and Koch‐Weser, J. 1963. Physical factors in the analysis of the actions of drugs on myocardial contractility. Pharmacol. Rev. 15:531‐599.
   Kaumann, A.J. 1981. In kitten ventricular myocardium, the inotropic potency of an agonist is determined by both its intrinsic activity for the adenylyl cyclase and its affinity for the β‐adrenoceptors. Naunyn‐Schmiedebergs Arch. Pharmakol. 317:13‐18.
   Koch‐Weser, J. and Blinks, J.R. 1963. The influence of the interval between beats on myocardial contractility. Pharmacol.Rev. 15:601‐652.
   Sonnenblick, E.H. 1962a. Force‐velocity relationships in mammalian heart muscle. Am. J. Physiol. 202:931‐939.
   Sonnenblick, E.H. 1962b. Implications of muscle mechanics in the heart. Fed. Proc. 21:975‐990.
   Sonnenblick, E.H. and McCallum, Z.T. 1961. Active state, force velocity relationships and inotropic mechanisms in mammalian papillary muscle. Fed. Proc. 20:126‐134.
Key References
   Blinks, J.R. 1966. Field stimulation as a means of effecting the release of autonomic transmitters in isolated heart muscle. J. Pharmacol. Exp. Ther. 151:221‐235.
  Describes the relationship between electrical stimulation and release of endogenous catecholamines in isolated heart muscle.
   Blinks and Koch‐Weser, 1963. See above.
  This pair of papers details the physical factors involved in the care and treatment of isolated heart muscle.
   Koch‐Weser and Blinks, 1963. See above.
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