Genomic and physiological analyses of the zebrafish atrioventricular canal reveal molecular building blocks of the secondary pacemaker region

• DOI: 10.1007/s00018-021-03939-y
• Publication Date: 2021-09-24T03:17:58Z
• Authors: Karim Abu Nahia; Maciej Migdał; T. Alexander Quinn; Kar-Lai Poon; Maciej Łapiński; Agata Sulej; Jiandong Liu; Shamba S. Mondal; Michał Pawlak; Łukasz Bugajski; Katarzyna Piwocka; Thomas Brand; Peter Kohl; Vladimir Korzh; Cecilia Winata
• Keywords: 0301 basic medicine; Biochemistry & Molecular Biology; Pacemaker, Artificial; Embryo, Nonmammalian; Organogenesis; Atrioventricular canal; IMPULSE PROPAGATION; 610; Atrioventricular node; 0601 Biochemistry and Cell Biology; Animals, Genetically Modified; 03 medical and health sciences; CARDIAC CONDUCTION; Animals; Wnt Signaling Pathway; IN-VIVO; Zebrafish; Science & Technology; Genome; Cardiac pacemaker; Heart Septal Defects; Myocardium; Gene Expression Regulation, Developmental; 1103 Clinical Sciences; Cell Biology; Genomics; Zebrafish Proteins; 0606 Physiology; EPITHELIAL-MESENCHYMAL TRANSITION; Heart Valves; OUTFLOW TRACT; DIFFERENTIATION; ENHANCER TRAP; Cardiac valve; JUNCTION; PROTEIN ISL-1; HEART; Original Article; RNA-seq; Life Sciences & Biomedicine

AbstractThe atrioventricular canal (AVC) is the site where key structures responsible for functional division between heart regions are established, most importantly, the atrioventricular (AV) conduction system and cardiac valves. To elucidate the mechanism underlying AVC development and function, we utilized transgenic zebrafish line sqet31Et expressing EGFP in the AVC to isolate this cell population and profile its transcriptome at 48 and 72 hpf. The zebrafish AVC transcriptome exhibits hallmarks of mammalian AV node, including the expression of genes implicated in its development and those encoding connexins forming low conductance gap junctions. Transcriptome analysis uncovered protein-coding and noncoding transcripts enriched in AVC, which have not been previously associated with this structure, as well as dynamic expression of epithelial-to-mesenchymal transition markers and components of TGF-β, Notch, and Wnt signaling pathways likely reflecting ongoing AVC and valve development. Using transgenic line Tg(myl7:mermaid) encoding voltage-sensitive fluorescent protein, we show that abolishing the pacemaker-containing sinoatrial ring (SAR) through Isl1 loss of function resulted in spontaneous activation in the AVC region, suggesting that it possesses inherent automaticity although insufficient to replace the SAR. The SAR and AVC transcriptomes express partially overlapping species of ion channels and gap junction proteins, reflecting their distinct roles. Besides identifying conserved aspects between zebrafish and mammalian conduction systems, our results established molecular hallmarks of the developing AVC which underlies its role in structural and electrophysiological separation between heart chambers. This data constitutes a valuable resource for studying AVC development and function, and identification of novel candidate genes implicated in these processes.