A multi-endpoint in vivo larval zebrafish (Danio rerio) model for the assessment of integrated cardiovascular function

• Published in: Journal of pharmacological and toxicological methods 2014-01, Vol.69 (1), p.30-38
• Main Authors: Parker, Thomas, Libourel, Paul-Antoine, Hetheridge, Malcolm J., Cumming, Robert I., Sutcliffe, Thomas P., Goonesinghe, Alexander C., Ball, Jonathan S., Owen, Stewart F., Chomis, Yann, Winter, Matthew J.
• Format: Article
• Language: English
• Subjects: Alternative animal model; Animals; Cardiovascular Agents - adverse effects; Cardiovascular Agents - pharmacology; Cardiovascular function; Cardiovascular pharmacology; Cardiovascular Physiological Phenomena - drug effects; Cardiovascular System - drug effects; Cardiovascular System - physiopathology; Danio rerio; Drug discovery; Drug safety assessment; Frontloading; Larva - drug effects; Larval zebrafish; Method; Models, Animal; Regional Blood Flow - drug effects; Regional Blood Flow - physiology; Zebrafish - physiology
• ISSN: 1056-8719; 1873-488X
• DOI: 10.1016/j.vascn.2013.10.002

Despite effective in vitro preclinical strategies to identify cardiovascular (CV) liabilities, there remains a need for early functional assessment prior to complex in vivo mammalian models. The larval zebrafish (Danio rerio, Zf) has been suggested for this role: previous data suggest that cardiac electrophysiology and vascular ultrastructure are comparable with mammals, and also indicate responsiveness of individual Zf CV system endpoints to some functional modulators. Little information is, however, available regarding integrated functional CV responses to drug treatment. Consequently, we developed a novel larval Zf model capable of simultaneous quantification of chronotropic, inotropic and arrhythmic effects, alongside measures of blood flow and vessel diameter. Non-invasive video analysis of the heart and dorsal aorta of anaesthetized and agarose-embedded larval ZF was used to measure multiple cardiovascular endpoints, simultaneously, following treatment with a range of functional modulators of CV physiology. Changes in atrial and ventricular beat frequencies were detected in response to acute treatment with cardio-stimulants (adrenaline and theophylline), and negative chrono/inotropes (cisapride, haloperidol, terfenadine and verapamil). Arrhythmias were also observed including terfenadine-induced 2:1 atrial–ventricular (A–V) block, a previously proposed hERG surrogate measure. Significant increases in blood flow were detected in response to adrenaline and theophylline exposure; and decreases after cisapride, haloperidol, terfenadine, and verapamil treatment. Using dorsal aorta (DA) blood flow and ventricular beat rate, surrogate stoke volumes were also calculated for all compounds. These data support the use of this approach for CV function studies. Moreover the throughput and compound requirements (approximately 3 compounds/person effort/week and