Presentation description

Atrial Fibrillation (AF), the most prevalent type of heart arrhythmia, is projected to affect 12.1 million individuals by 2030. Our previous work has linked a mutation in the nuclear factor of activated cell 1 (NFATc1) protein to an increased susceptibility to develop young-onset AF (<40 yo) in a Utah family. NFATc1 is a calcium-dependent transcription factor expressed in the heart of several species, including zebrafish. We are now using the zebrafish heart as a model to elucidate the mechanism behind this increased susceptibility. Our objective is to identify a suitable panel of antibodies to establish the specific cell type in which NFATc1 is expressed in the zebrafish heart. These antibodies must be compatible with the iDISCO clearing technique for comprehensive visualization of the entire zebrafish heart. To accomplish this, we have employed standard immunohistochemical techniques and confocal microscopy to acquire an image stack, which we subsequently processed using the open-source NIH software Fiji and IMARIS. We have successfully identified two antibodies that are compatible with our clearing technique: an anti-myosin heavy chain antibody (DSHB, MF20) to identify cardiomyocytes and an anti- vimentin antibody (DSHB, H5) to identify fibroblasts. Future studies will involve testing antibodies against other cell types present in the heart and optimizing an antibody that can selectively reveal the localization of NFATc1. Identifying NFATc1 expression in specific cell types is the first step to elucidating the possible role of NFATc1 in the development of an atrial fibrillation phenotype, providing mechanistic insight into this disease and its potential future treatments