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Methods for the Isolation, Culture, and Functional Characterization of Sinoatrial Node Myocytes from Adult Mice
Published on: October 23, 2016
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RHOA-ROCK signalling is necessary for lateralization and differentiation of the developing sinoatrial node
Rebecca Vicente-Steijn1,2,3, Tim P Kelder1, Leon G Tertoolen1
1Department of Anatomy & Embryology, Leiden University Medical Center, Leiden, The Netherlands.
Cardiovascular Research
|September 14, 2017
Summary
RHOA-ROCK signaling is crucial for developing the heart's primary pacemaker, the sinoatrial node (SAN). Inhibiting this pathway in embryonic chickens prolonged immature states and caused developmental abnormalities, highlighting its role in SAN specialization.
Area of Science:
- Cardiovascular Development
- Molecular Cardiology
- Developmental Biology
Background:
- RHOA-ROCK signaling influences cell functions like migration and proliferation.
- RHOA is present in the developing cardiac conduction system.
- Pacemaker potential is initially widespread but later restricted to the right sinoatrial node (SAN).
Purpose of the Study:
- To investigate the role of RHOA-ROCK signaling in the development of the SAN in chicken embryos.
- To understand how RHOA-ROCK signaling contributes to the specialization of the SAN.
Main Methods:
- Chemical inhibition of ROCK signaling using Y-27632 in embryonic chicken hearts.
- Analysis of gene expression, including transcription factors and SAN-specific genes.
- Electrophysiological assessment using patch clamp techniques.
Main Results:
- ROCK inhibition prolonged the immature state of the sinus venosus myocardium, indicated by ISL1 upregulation and widespread pacemaker potential.
- Inhibition led to aberrant expression of key SAN and left-right asymmetry genes.
- Electrophysiology confirmed an immature SAN phenotype and residual pacemaker potential on the left side.
Conclusions:
- RHOA-ROCK signaling is essential for establishing the right-sided SAN as the definitive cardiac pacemaker.
- This signaling pathway restricts the expression of pacemaker genes to the right sinus venosus myocardium, ensuring proper SAN function.
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