Automatic Zebrafish Heartbeat Detection and Analysis for Zebrafish Embryos

Pylatiuk, Christian; Sánchez, Daniela; Mikut, Ralf; Alshut, Ruediger; Reischl, Markus; Hirth, Sofia; Rottbauer, Wolfgang; Just, Steffen

doi:10.1089/zeb.2014.1002

Cited by 58 publications

(41 citation statements)

References 20 publications

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“…The rather dramatic baseline difference in heart rate between F0 and the other generations was potentially due to consistent overestimation by the F0 technique of visual counting and extrapolating and the temperature variability (Barrionuevo and Burggren, 1999). Adoption of a digital motion analysis technique by the time the F2 measurements were made obviated reliance on extrapolation (Pylatiuk et al , 2014). …”

Section: Resultsmentioning

confidence: 99%

Transgenerational inheritance of neurobehavioral and physiological deficits from developmental exposure to benzo[a]pyrene in zebrafish

Knecht

Truong

Marvel

et al. 2017

Toxicology and Applied Pharmacology

118

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Benzo[a]pyrene (B[a]P) is a well-known genotoxic polycylic aromatic compound whose toxicity is dependent on signaling via the aryl hydrocarbon receptor (AHR). It is unclear to what extent detrimental effects of B[a]P exposures might impact future generations and whether transgenerational effects might be AHR-dependent. This study examined the effects of developmental B[a]P exposure on 3 generations of zebrafish. Zebrafish embryos were exposed from 6 – 120 hours post fertilization (hpf) to 5 and 10 μM B[a]P and raised in chemical- free water until adulthood (F0). Two generations were raised from F0 fish to evaluate transgenerational inheritance. Morphological, physiological and neurobehavioral parameters were measured at two life stages. Juveniles of the F0 and F2 exhibited hyper locomotor activity, decreased heartbeat and mitochondrial function. B[a]P exposure during development resulted in decreased global DNA methylation levels and generally reduced expression of DNA methyltransferases in wild type zebrafish, with the latter effect largely reversed in an AHR2-null background. Adults from the F0 B[a]P exposed lineage displayed social anxiety-like behavior. Adults in the F2 transgeneration manifested gender-specific increased body mass index (BMI), increased oxygen consumption and hyper-avoidance behavior. Exposure to benzo[a]pyrene during development resulted in transgenerational inheritance of neurobehavioral and physiological deficiencies. Indirect evidence suggested the potential for an AHR2-dependent epigenetic route.

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Section: Resultsmentioning

confidence: 99%

Transgenerational inheritance of neurobehavioral and physiological deficits from developmental exposure to benzo[a]pyrene in zebrafish

Knecht

Truong

Marvel

et al. 2017

Toxicology and Applied Pharmacology

118

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“…Thus, the use of the zebrafish as a model in these studies will facilitate more extensive, easy and comprehensive knowledge of drugs cardiotoxicity, generating a deeper understanding of that process. Similarly, the zebrafish is also an attractive screening tool for cardiovascular risk assessment after treatment with atypical antipsychotic drugs, as it facilitates the evaluation of the heart beat rate [66] .…”

Section: Antipsychotics Toxicity Testingmentioning

confidence: 99%

Zebrafish as Toxicological model for screening and recapitulate human diseases

Caballero¹,

Candiracci²

2018

JUMD

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Embryonic and larval Danio rerio (zebrafish) is increasingly used as a toxicological model to conduct rapid in vivo tests and developmental toxicity assays; the zebrafish features high genetic homology to mammals, robust, phenotypes, highthroughput genetic and chemical screening have made it a powerful tool to evaluate in vivo toxicity. New methodologies of genome editing as CRISPR/Cas9; ZFN and TALEN make it a suitable model to perform studies to pair human genetic diseases as well. This review surveys recent studies employing zebrafish as experimental model, comparing it with other in vivo and in vitro models, presenting zebrafish as a potent vertebrate tool to evaluate drug toxicity and efficacy in order to facilitate more extensive, easy and comprehensive knowledge of new generation drugs.

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“…To date, several methods have been used to assess heartbeat in zebrafish hatchlings, medaka embryos, fish species, Drosophila, cardiomyocytes or mouse embryos 23,24,26,27,29,30,[39][40][41][42][43] . Some of these methods require transgenic embryos with fluorescent cardiomyocytes or erythrocytes to measure heartbeats 28,39 .…”

Section: Discussionmentioning

confidence: 99%

“…Readouts of heart rate and rhythm have been employed increasingly in zebrafish-and medaka-based screens [23][24][25][26][27][28][29][30] . Challenges of current methods to capture heart rate in high-throughput include: (i) immobilization of dechorionated embryos or hatchlings, which in the majority of cases is achieved by anesthesia with tricaine leading to severe adverse cardiovascular effects 31 , (ii) imaging of larvae in standardized orientations, involving labor-intensive manual mounting on dedicated sample carriers or embedding in low-melting agarose, (iii) fluorescent reporter readouts, which preclude studies of specific genetic backgrounds and (iv) lack of user-friendly software for efficient analysis of multiple hearts simultaneously.…”

Section: Introductionmentioning

confidence: 99%

Automated high-throughput heart rate measurement in medaka and zebrafish embryos under physiological conditions

Gierten

Pylatiuk

Hammouda

et al. 2019

Preprint

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Rationale: Accurate and efficient quantification of heartbeats in small fish models is an important readout to study cardiovascular biology, disease states and pharmacology at large scale. However, dependence on anesthesia, laborious sample orientation or requirement for fluorescent reporters have hampered the establishment of high-throughput heartbeat analysis.Objective: To overcome these limitations, we aimed to develop a high-throughput assay with automated heart rate scoring in medaka (Oryzias latipes) and zebrafish (Danio rerio) embryos under physiological conditions designed for genetic screens and drug discovery and validation. Methods and Results:We established an efficient screening assay employing automated label-free heart rate determination of randomly oriented, non-anesthetized specimen in microtiter plates. Automatically acquired bright-field data feeds into an easy-to-use HeartBeat software, a MATLAB algorithm with graphical user interface developed for automated quantification of heart rate and rhythm. Sensitivity of the assay and algorithm was demonstrated by profiling heart rates during entire embryonic development. Our analysis pipeline revealed acute temperature changes triggering rapid adaption of heart rates, which has implications for standardization of experimental layout. The approach is scalable and allows scoring of multiple embryos per well resulting in a throughput of >500 embryos per 96-well plate. In a proof of principle screen for compound testing, our assay captured concentration-dependent effects of nifedipine and terfenadine over time. Conclusion:A novel workflow and HeartBeat software provide efficient means for reliable and direct quantification of heart rate and rhythm of small fish in a physiological environment. Importantly, confounding factors such as anesthetics or laborious mounting are eliminated. We provide detailed profiles of embryonic heart rate dynamics in medaka and zebrafish as reference for future assay development. Ease of sample handling, automated imaging, physiological conditions and softwareassisted analysis now facilitate various large-scale applications ranging from phenotypic screening, interrogation of gene functions to cardiovascular drug development pipelines.

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Automatic Zebrafish Heartbeat Detection and Analysis for Zebrafish Embryos

Cited by 58 publications

References 20 publications

Transgenerational inheritance of neurobehavioral and physiological deficits from developmental exposure to benzo[a]pyrene in zebrafish

Transgenerational inheritance of neurobehavioral and physiological deficits from developmental exposure to benzo[a]pyrene in zebrafish

Zebrafish as Toxicological model for screening and recapitulate human diseases

Automated high-throughput heart rate measurement in medaka and zebrafish embryos under physiological conditions

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