Quantification of larval zebrafish motor function in multiwell plates using open-source MATLAB applications

Zhou, Yangzhong; Cattley, Richard T.; Cario, Clinton L.; Bai, Quan; Burton, Edward A.

doi:10.1038/nprot.2014.094

Cited by 49 publications

(42 citation statements)

References 17 publications

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“…At 5 days post-fertilization (dpf), zebrafish larvae were transferred to 96-well plates in fresh E3 buffer and allowed to acclimatize inside a recording incubator for 30 minutes before experiments started. Video recordings of larvae swimming in the wells of the 96-well plate were captured at 4 frames/s under infrared illumination and analyzed as described in our previous work [4, 19]. …”

Section: Methodsmentioning

confidence: 99%

Spectral properties of the zebrafish visual motor response

Burton

Zhou

Bai

et al. 2017

Neuroscience Letters

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Larval zebrafish react to changes in ambient illumination with a series of stereotyped motor responses, called the visual motor response (VMR). The VMR has been used widely in zebrafish models to analyze how genetic or environmental manipulations alter neurological function. Prior studies elicited the VMR using white light. In order to elucidate the underlying afferent pathways and to identify light wavelengths that elicit the VMR without also activating optogenetic reagents, we employed calibrated narrow-waveband light sources to analyze the spectral properties of the response. Narrow light wavebands with peaks between 399nm and 632nm triggered the characteristic phases of the VMR, but there were quantitative differences between responses to different light wavelengths at the same irradiant flux density. The O-bend component of the VMR was elicited readily at dark onset following illumination in 399nm or 458nm light, but was less prominent at the transition from 632nm light to dark. Conversely, stable motor activity in light was observed at 458nm, 514nm, and 632nm, but not at 399nm. The differential effect of discrete light wavebands on components of the VMR suggests they are driven by distinct photoreceptor populations. Furthermore, these data enable the selection of light wavebands to drive the VMR in a separate channel to the activation of optogenetic reagents and photosensitizers.

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Section: Methodsmentioning

confidence: 99%

Spectral properties of the zebrafish visual motor response

Burton

Zhou

Bai

et al. 2017

Neuroscience Letters

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“…Zebrafish locomotor behavioral testing was performed exactly as described in our previous work (Farrell et al, 2011; Zhou et al, 2014). Briefly, 5dpf zebrafish larvae were placed in the wells of a black-walled 96-well plate with an optical glass bottom, and imaged in a 28.5°C incubator under bright white illumination (brightness 200 Lux, color temperature 3500K).…”

Section: Methodsmentioning

confidence: 99%

“…The larvae were then transferred to individual wells of a 96-well plate in fresh medium, and their movements measured by analysis of a video stream (Cario et al, 2011; Zhou et al, 2014). (C) Representative vector plot showing the centroid locus of each zebrafish in 1000 consecutive video frames at 4 frames/sec.…”

Section: Figurementioning

confidence: 99%

Live imaging of mitochondrial dynamics in CNS dopaminergic neurons in vivo demonstrates early reversal of mitochondrial transport following MPP+ exposure

Dukes

Bai

Laar

et al. 2016

Neurobiology of Disease

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Extensive convergent evidence collectively suggests that mitochondrial dysfunction is central to the pathogenesis of Parkinson’s disease (PD). Recently, changes in the dynamic properties of mitochondria have been increasingly implicated as a key proximate mechanism underlying neurodegeneration. However, studies have been limited by the lack of a model in which mitochondria can be imaged directly and dynamically in dopaminergic neurons of the intact vertebrate CNS. We generated transgenic zebrafish in which mitochondria of dopaminergic neurons are labeled with a fluorescent reporter, and optimized methods allowing direct intravital imaging of CNS dopaminergic axons and measurement of mitochondrial transport in vivo. The proportion of mitochondria undergoing axonal transport in dopaminergic neurons decreased overall during development between 2 days post-fertilization (dpf) and 5dpf, at which point the major period of growth and synaptogenesis of the relevant axonal projections is complete. Exposure to 0.5 – 1.0mM MPP+ between 4 – 5 dpf did not compromise zebrafish viability or cause detectable changes in the number or morphology of dopaminergic neurons, motor function or monoaminergic neurochemistry. However, 0.5mM MPP+ caused a 300% increase in retrograde mitochondrial transport and a 30% decrease in anterograde transport. In contrast, exposure to higher concentrations of MPP+ caused an overall reduction in mitochondrial transport. This is the first time mitochondrial transport has been observed directly in CNS dopaminergic neurons of a living vertebrate and quantified in a PD model in vivo. Our findings are compatible with a model in which damage at presynaptic dopaminergic terminals causes an early compensatory increase in retrograde transport of compromised mitochondria for degradation in the cell body. These data are important because manipulation of early pathogenic mechanisms might be a valid therapeutic approach to PD. The novel transgenic lines and methods we developed will be useful for future studies on mitochondrial dynamics in health and disease.

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“…Behavioral phenotypes are the most complex manifestations of multiple diseases, especially for those affecting the CNS or musculoskeletal systems (Stewart et al 2014). Automated behavioral video tracking systems have been developed to quantify the fish behavior with a range of parameters on a large scale (Zhou et al 2014). Corresponding functional behavioral tests and experimental set-ups are gradually standardized for both larval and adult zebrafish, proving the potential of zebrafish in cost-effective high-throughput screens (HTSs) and therapy development (Rihel et al 2010) (Fig.…”

Section: Zebrafish: a Prepared Model For Studying Human Pathogenesismentioning

confidence: 99%

CRISPR/Cas9 in zebrafish: an efficient combination for human genetic diseases modeling

Liu

Zhou

et al. 2016

Hum Genet

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The next-generation sequencing identifies a growing number of candidate genes associated with human genetic diseases, which inevitably requires efficient methods to validate the causal links between genotype and phenotype. Recently, zebrafish, with sufficiently high-throughput capabilities, has become a favored option to study human pathogenesis. In addition, CRISPR/Cas9-based approaches have radically reduced the efforts to introduce targeted genome engineering in various organisms. Here, we systemically review the basic considerations in the design of gene editing in zebrafish with CRISPR/Cas9, and explore the potential of the combination of these two to support efficient functional analysis of human genetic variants.

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Quantification of larval zebrafish motor function in multiwell plates using open-source MATLAB applications

Cited by 49 publications

References 17 publications

Spectral properties of the zebrafish visual motor response

Spectral properties of the zebrafish visual motor response

Live imaging of mitochondrial dynamics in CNS dopaminergic neurons in vivo demonstrates early reversal of mitochondrial transport following MPP+ exposure

CRISPR/Cas9 in zebrafish: an efficient combination for human genetic diseases modeling

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