Abstract
Zebrafish is a widespread used model in cardiovascular research, and recently adult zebrafish, as a more human relevant model compared to larval stages, became particularly valuable to study heart regeneration. While cardiac performance in larval stages can be easily assessed using transillumination microscopy, methods to reliably assess cardiac function in adult zebrafish are largely missing. Here, we developed the first standardized protocol to reliably assess cardiac function in adult zebrafish using high frequency echocardiography and speckle-tracking algorithms. Respecting distinct anatomic characteristics in zebrafish, we defined three examination planes according to standards in human echocardiography. Further, by combining conventional echocardiographic measurements with modern speckle tracking based strain analysis we are able to measure myocardial performance in extremely high spatial and temporal resolution. Thereby we were able to attain high quality B-Mode imaging and PW-Doppler signals allowing distinct detection of changes in chronotropy, inotropy and, for the first time in adult zebrafish, in dromotropy in response to treatment with Atenolol and Isoproterenol. Additionally, we employed zebrafish that underwent cardiac cryoinjury to longitudinally follow functional cardiac regeneration. High-resolution speckle-tracking strain analysis allowed us to demarcate injured myocardial regions and demonstrate that functional healing extends beyond 30-45 days post injury (dpi) as previously implied by conventional methodologies. Specifically, we found delayed restitution of cardiac displacement and re-synchronization of injured and non-injured myocardium beyond 60 dpi until 120 dpi. In summary, our protocol enables highly reproducible and high throughput measurements of cardiac performance in adult zebrafish in a high spatio-temporal resolution. Thereby, our protocol represents a valuable novel tool for functional cardiac assessment and to detect changes in cardiac function in response to gene mutation or myocardial injury and might lead to further insights into cardiac physiology and disease.
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