title: A multi-endpoint in vivo larval zebrafish (Danio rerio) model for the assessment of integrated cardiovascular function.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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 <10 mg) make our approach potentially suitable for higher throughput drug safety and efficacy applications, pending further assessment of ZF-mammalian pharmacological comparability.
authors.affiliation: AstraZeneca Global Safety, Health and Environment, Brixham Environmental Laboratory, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Thomas
authors.initials: T
authors.lastname: Parker
authors.affiliation: Centre de Recherche en Neurosciences de Lyon, Faculté de Médecine R.T.H. Laennec, 7 rue Guillaume Paradin, 69372 Lyon, France.
authors.forename: Paul-Antoine
authors.initials: PA
authors.lastname: Libourel
authors.affiliation: AstraZeneca Global Safety, Health and Environment, Brixham Environmental Laboratory, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: Hetheridge
authors.affiliation: AstraZeneca Global Safety, Health and Environment, Brixham Environmental Laboratory, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Robert I
authors.initials: RI
authors.lastname: Cumming
authors.affiliation: AstraZeneca Global Safety, Health and Environment, Brixham Environmental Laboratory, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Sutcliffe
authors.affiliation: AstraZeneca Global Safety, Health and Environment, Brixham Environmental Laboratory, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Alexander C
authors.initials: AC
authors.lastname: Goonesinghe
authors.affiliation: AstraZeneca Global Safety, Health and Environment, Brixham Environmental Laboratory, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Ball
authors.affiliation: AstraZeneca Global Safety, Health and Environment, Brixham Environmental Laboratory, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom.
authors.forename: Stewart F
authors.initials: SF
authors.lastname: Owen
authors.affiliation: ViewPoint, 3 allée des Chevreuils, F-69380 Lissieu, France.
authors.forename: Yann
authors.initials: Y
authors.lastname: Chomis
authors.affiliation: AstraZeneca Global Safety, Health and Environment, Brixham Environmental Laboratory, Freshwater Quarry, Brixham, Devon TQ5 8BA, United Kingdom. Electronic address: matthew.winter@astrazeneca.com.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Winter
mesh_terms: D000818:Animals; D002317:Cardiovascular Agents / Q000009:adverse effects* / Q000494:pharmacology*; D002320:Cardiovascular Physiological Phenomena / Q000187:drug effects*; D002319:Cardiovascular System / Q000187:drug effects* / Q000503:physiopathology*; D007814:Larva / Q000187:drug effects; D023421:Models, Animal; D012039:Regional Blood Flow / Q000187:drug effects / Q000502:physiology; D015027:Zebrafish / Q000502:physiology*
chemical_list: D002317:Cardiovascular Agents
keywords: ABR; AR; Alternative animal model; A–V; BP; CV; Cardiovascular function; Cardiovascular pharmacology; DMSO; Drug discovery; Drug safety assessment; Frontloading; L-NAME; Larval zebrafish; MS222; MTC; Method; N (G)-nitro-l-arginine methyl ester; SC; SEM; SNP; SSV; VBR; VSMCs; Zf; adrenoreceptor; atrial beat rate; atrial–ventricular; beats per minute; blood pressure; bpm; cardiovascular; days post fertilization; dimethyl sulphoxide; dpf; fps; frames per second; hour post fertilization; hpf; maximum tolerated concentration; sodium nitroprusside; solvent control; standard error of the mean; surrogate stroke volume; tricaine methanesulfonate; vascular smooth muscle cells; ventricular beat rate; zebrafish