In Vitro Assessment of Cardiac Function Using Skinned Cardiomyocytes

Data de publicação: 01/06/2020 Data Ahead of Print: 22/06/2020

Autores da FMUP

• Joaquim Adelino Correia Ferreira Leite Moreira

  Autor

• Inês Maria Falcão Sousa Pires Marques

  Autor

Participantes de fora da FMUP

• Gonçalves-Rodrigues, P
• Almeida Coelho, J
• Gonçalves, A
• Amorim, F
• Stienen, GJM

Unidades de investigação

• Cirurgia e Fisiologia

• RISE-Health

  

  Investigador

  Fernando Carlos De Landér Schmitt

Abstract

In this article, we describe the steps required to isolate a single permeabilized ("skinned") cardiomyocyte and attach it to a force-measuring apparatus and a motor to perform functional studies. These studies will allow measurement of cardiomyocyte stiffness (passive force) and its activation with different calcium (Ca2+)-containing solutions to determine, amongst others: maximum force development, myofilament Ca2+-sensitivity (pCa(50)), cooperativity (nHill) and the rate of force redevelopment (ktr). This method also enables determination of the effects of drugs acting directly on myofilaments and of the expression of exogenous recombinant proteins on both active and passive properties of cardiomyocytes. Clinically, skinned cardiomyocyte studies highlight the pathophysiology of many myocardial diseases and allow in vitro assessment of the impact of therapeutic interventions targeting the myofilaments. Altogether, this technique enables the clarification of cardiac pathophysiology by investigating correlations between in vitro and in vivo parameters in animal models and human tissue obtained during open heart or transplant surgery.

Keywords

• Actin Cytoskeleton; Animals; Calcium; Cardiovascular Physiological Phenomena; Mice; Myocardial Contraction; Myocytes, Cardiac; Myofibrils; calcium; actin filament; animal; cardiac muscle cell; cardiovascular function; cytology; heart contraction; metabolism; mouse; muscle fibril; physiology