An optimized method for counting dopaminergic neurons in zebrafish

Matsui, Hirokazu; Sugie, Atsushi

doi:10.1371/journal.pone.0184363

Cited by 28 publications

(22 citation statements)

References 45 publications

(59 reference statements)

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“…To study the regeneration of distinct populations of neurons without physical damage, we ablated dopaminergic and noradrenergic neurons using 6-hydroxydopamine (6-OHDA), which selectively ablates these neurons across vertebrates (Berg et al, 2011;Tieu, 2011;Matsui and Sugie, 2017;Vijayanathan et al, 2017). In adult zebrafish, the dopaminergic system is highly differentiated.…”

Section: Introductionmentioning

confidence: 99%

Regeneration of Dopaminergic Neurons in Adult Zebrafish Depends on Immune System Activation and Differs for Distinct Populations

et al. 2019

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Adult zebrafish, in contrast to mammals, regenerate neurons in their brain, but the extent and variability of this capacity is unclear. Here we ask whether the loss of various dopaminergic neuron populations is sufficient to trigger their functional regeneration. Both sexes of zebrafish were analyzed. Genetic lineage tracing shows that specific diencephalic ependymo-radial glial (ERG) progenitor cells give rise to new dopaminergic [tyrosine hydroxylase-positive (TH ϩ)] neurons. Ablation elicits an immune response, increased proliferation of ERG progenitor cells, and increased addition of new TH ϩ neurons in populations that constitutively add new neurons (e.g., diencephalic population 5/6). Inhibiting the immune response attenuates neurogenesis to control levels. Boosting the immune response enhances ERG proliferation, but not addition of TH ϩ neurons. In contrast, in populations in which constitutive neurogenesis is undetectable (e.g., the posterior tuberculum and locus ceruleus), cell replacement and tissue integration are incomplete and transient. This is associated with a loss of spinal TH ϩ axons, as well as permanent deficits in shoaling and reproductive behavior. Hence, dopaminergic neuron populations in the adult zebrafish brain show vast differences in regenerative capacity that correlate with constitutive addition of neurons and depend on immune system activation.

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Section: Introductionmentioning

confidence: 99%

Regeneration of Dopaminergic Neurons in Adult Zebrafish Depends on Immune System Activation and Differs for Distinct Populations

et al. 2019

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“…To study regeneration of distinct populations of neurons without physical damage, we ablated dopaminergic and noradrenergic neurons using 6-hydroxydopamine (6OHDA), which selectively ablates these neurons across vertebrates [6][7][8][9] . In adult zebrafish, the dopaminergic system is highly differentiated.…”

Section: Introductionmentioning

confidence: 99%

Regeneration of dopaminergic neurons in adult zebrafish depends on immune system activation and differs for distinct populations

Caldwell¹,

Davies²,

Cavone³

et al. 2018

Preprint

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Adult zebrafish regenerate neurons in their brain, but the extent and variability of this capacity is unclear. Here we ask whether loss of various dopaminergic neuron populations is sufficient to trigger their functional regeneration. Genetic lineage tracing shows that specific diencephalic ependymo-radial glial progenitor cells (ERGs) give rise to new dopaminergic (Th+) neurons. Ablation elicits an immune response, increased proliferation of ERGs and increased addition of new Th+ neurons in populations that constitutively add new neurons, e.g. diencephalic population 5/6. Inhibiting the immune response attenuates neurogenesis to control levels. Boosting the immune response enhances ERG proliferation, but not addition of Th+ neurons. In contrast, in populations in which constitutive neurogenesis is undetectable, e.g. the posterior tuberculum and locus coeruleus, cell replacement and tissue integration are incomplete and transient. This is associated with loss of spinal Th+ axons, as well as permanent deficits in shoaling and reproductive behaviour. Hence, dopaminergic neuron populations in the adult zebrafish brain show vast differences in regenerative capacity that correlate with constitutive addition of neurons and depend on immune system activation.

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“…To investigate whether these cells are impacted in our zebrafish models of PD, we used immunofluorescence to detect the protein Tyrosine hydroxylase (Th), an enzyme required to produce catecholamines including dopamine. Our method is informed by the work of Matsui and Sugie (2017) and Rink and Wullimann (2001) to identify and count these neurons in zebrafish. The brains were fixed and embedded in agarose blocks for vibratome sectioning before processing by immunohistochemistry for Th immunoreactivity and imaging by confocal microscopy.…”

Section: Resultsmentioning

confidence: 99%

“…Brains were then sectioned transversely in 3% agarose/PBS by vibratome at a thickness of 100μM. Based on the protocol by (Matsui and Sugie, 2017) brains were incubated in 10 mM sodium citrate buffer (pH 8.5) for 2 hours at 80°C. Sections were washed twice in PBS with 1% TritonX-100 (PBS/1% Tx) for 15 minutes.…”

Section: Immunofluorescencementioning

confidence: 99%

Machine learning discriminates a movement disorder in a zebrafish model of Parkinson's disease

Hughes

Lones

Bedder

et al. 2020

Disease Models &Amp; Mechanisms

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Animal models of human disease provide an in vivo system that can reveal molecular mechanisms by which mutations cause pathology, and, moreover, have the potential to provide a valuable tool for drug development. Here we have developed a zebrafish model of Parkinson's disease (PD) together with a novel method to screen for movement disorders in adult fish, pioneering a more efficient drug testing route. Mutation of the PARK7 gene (DJ-1) is known to cause monogenic autosomal recessive PD in humans and, using CRISPR/Cas9 gene editing we have generated a DJ-1 loss of function zebrafish with molecular hallmarks of PD. To establish whether there is a human-relevant parkinsonian phenotype in our model, we have adapted proven tools used to diagnose PD in clinics and have developed a novel and unbiased computational method to classify movement disorders in adult zebrafish. Using high-resolution video capture and machine learning, we have extracted novel features of movement from continuous data streams and used an evolutionary algorithm (EA) to classify parkinsonian fish. This method will be widely applicable for assessing zebrafish models of human motor diseases and provide a valuable asset for the therapeutics pipeline. In addition, interrogation of RNA-seq data indicate metabolic reprogramming of brains in the absence of DJ-1, adding to growing evidence that disruption of bioenergetics is a key feature of neurodegeneration.

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An optimized method for counting dopaminergic neurons in zebrafish

Cited by 28 publications

References 45 publications

Regeneration of Dopaminergic Neurons in Adult Zebrafish Depends on Immune System Activation and Differs for Distinct Populations

Regeneration of Dopaminergic Neurons in Adult Zebrafish Depends on Immune System Activation and Differs for Distinct Populations

Regeneration of dopaminergic neurons in adult zebrafish depends on immune system activation and differs for distinct populations

Machine learning discriminates a movement disorder in a zebrafish model of Parkinson's disease

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