HCN4 Dynamically Marks the First Heart Field and Conduction System Precursors
Rationale: To date, there has been no specific marker of the first heart field to facilitate understandings of contributions of the first heart field to cardiac lineages. Cardiac arrhythmia is a leading cause of death, often resulting from abnormalities in the cardiac conduction system (CCS). Understanding origins and identifying markers of CCS lineages is an essential step toward modeling diseases of the CCS and for development of biological pacemakers.
Objective: To investigate HCN4 as a marker for the first heart field and for precursors of distinct components of the CCS, and gain insight into contributions of first and second heart lineages to the CCS.
Methods and Results: HCN4-CreERT2, -nuclear LacZ and -H2BGFP mouse lines were generated. HCN4 expression was examined by means of immunostaining with HCN4 antibody and reporter gene expression. Lineage studies were performed using HCN4CreERT2, Isl1Cre, Nkx2.5Cre, and Tbx18Cre, coupled to co-immunostaining with CCS markers. Results demonstrated that, at cardiac crescent stages, HCN4 marks the first heart field, with HCN4CreERT2 allowing assessment of cell fates adopted by first heart field myocytes. Throughout embryonic development, HCN4 expression marked distinct CCS precursors at distinct stages, marking the entire CCS by late fetal stages. We also noted expression of HCN4 in distinct subsets of endothelium at specific developmental stages.
Conclusions: This study provides insight into contributions of first and second heart lineages to the CCS and highlights the potential utility of HCN4 in conjunction with other markers for optimization of protocols for generation and isolation of specific conduction system precursors.
- cardiac lineage
- cardiac conduction system
- Heart field
- cardiac development
- cardiac arrhythmia
- transgenic model
- cardiac biomarkers
- Received April 13, 2013.
- Revision received June 5, 2013.
- Accepted June 6, 2013.