Characterization of cardiac β-adrenergic receptors in the guinea pig heart: Application to study of β-adrenergic receptors in shock models

Tina Kaufman, J. W. Horton

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14 Citations (Scopus)

Abstract

The myocardial response to catecholamines is significantly diminished in many types of shock or heart failure. The guinea pig heart is an ideal model for the study of shock, as it is relatively inexpensive, and the cardiovascular system of the guinea pig most closely resembles that of the human. Using this model, we have developed techniques to characterize and quantitate changes in β-adrenergic receptors (βAR) in the guinea pig heart after burn injury. Preliminary experiments were performed to determine the optimum binding conditions, e.g., incubation time and conditions, protein concentrations, rinsing, etc. Additional experiments were conducted using agonists and antagonists to characterize the rank order of potency and stereospecificity of the βAR. Crude membrane preparations (50 μg/250 μl) from sham-burned and burned hearts were incubated with 8-10 concentrations of 125I-cyanopindolol (10-450 pM) at 37°C for 1 hr. Under these conditions, binding assays were linear with respect to protein concentration and time. Alprenolol (10 μM) was used to determine nonspecific binding. The membrane preparations used in this study bound both agonists and antagonists with a rank order of potency and stereospecificity characteristic of a β-adrenergic receptor. Finally, agonist competition curves were performed with isoproterenol in the presence and absence of Gpp(NH)p to determine receptor regulation by the Gs protein. Analysis using computer-assisted techniques suggests that the fraction of high-affinity β-receptors is significantly reduced after burn injury (41.2 ± 4.7%) compared to sham-burned controls (54 ± 2%, P ≤ 0.023). The methods described here can be easily adapted to correlate cardiac dysfunction with changes in β-adrenergic receptors during many types of shock or trauma.

Original languageEnglish (US)
Pages (from-to)516-523
Number of pages8
JournalJournal of Surgical Research
Volume55
Issue number5
DOIs
StatePublished - 1993
Externally publishedYes

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Adrenergic Receptors
Shock
Guinea Pigs
Wounds and Injuries
Alprenolol
Guanylyl Imidodiphosphate
Proteins
Membranes
Cardiovascular System
Isoproterenol
Catecholamines
Heart Failure

ASJC Scopus subject areas

  • Surgery

Cite this

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abstract = "The myocardial response to catecholamines is significantly diminished in many types of shock or heart failure. The guinea pig heart is an ideal model for the study of shock, as it is relatively inexpensive, and the cardiovascular system of the guinea pig most closely resembles that of the human. Using this model, we have developed techniques to characterize and quantitate changes in β-adrenergic receptors (βAR) in the guinea pig heart after burn injury. Preliminary experiments were performed to determine the optimum binding conditions, e.g., incubation time and conditions, protein concentrations, rinsing, etc. Additional experiments were conducted using agonists and antagonists to characterize the rank order of potency and stereospecificity of the βAR. Crude membrane preparations (50 μg/250 μl) from sham-burned and burned hearts were incubated with 8-10 concentrations of 125I-cyanopindolol (10-450 pM) at 37°C for 1 hr. Under these conditions, binding assays were linear with respect to protein concentration and time. Alprenolol (10 μM) was used to determine nonspecific binding. The membrane preparations used in this study bound both agonists and antagonists with a rank order of potency and stereospecificity characteristic of a β-adrenergic receptor. Finally, agonist competition curves were performed with isoproterenol in the presence and absence of Gpp(NH)p to determine receptor regulation by the Gs protein. Analysis using computer-assisted techniques suggests that the fraction of high-affinity β-receptors is significantly reduced after burn injury (41.2 ± 4.7{\%}) compared to sham-burned controls (54 ± 2{\%}, P ≤ 0.023). The methods described here can be easily adapted to correlate cardiac dysfunction with changes in β-adrenergic receptors during many types of shock or trauma.",
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