Functional regionalization of the teleost cerebellum analyzed in vivo

Matsui, Hirokazu; Namikawa, Kazuhiko; Babaryka, Andreas; Köster, Reinhard W.

doi:10.1073/pnas.1403105111

Cited by 84 publications

(189 citation statements)

References 43 publications

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“…1B) (Kim et al, 2011). To achieve specific expression in differentiating cerebellar Purkinje cells (PCs), a regulatory element isolated from the upstream region of the zebrafish carbonic anhydrase 8 (ca8) homolog was used (Matsui et al, 2014), resulting in the stable transgenic zebrafish strain Tg(ca8:FMA-TagRFP-2A-casp8ERT2) bz11Tg (Fig. 1C) which we named PC-ATTAC TM , according to the previously used nomenclature (Pajvani et al, 2005) but emphasizing its tamoxifen dependence.…”

Section: Inducible Caspase Under Transient Transgenic Conditionsmentioning

confidence: 99%

“…Because tagRFP is fused at its N-terminus with the myristoylation site of the Fyn kinase, the axons, somata and the prominent dendrites of PCs could be observed in detail (Kameda et al, 2008), including the long axonal projections of PCs that form the cerebello-octavolateralis tract to vestibular nuclei (Bae et al, 2009;Matsui et al, 2014;Biechl et al, 2016) (Fig. 1F, white arrowheads).…”

Section: Inducible Caspase Under Transient Transgenic Conditionsmentioning

confidence: 99%

“…In addition, established transgenic lines expressing KalTA4 in the ventral neural tube (hzm9; Babaryka et al, 2009), in rhombomeres 3 and 5 (hzm6Et), and in skeletal muscles (hzm8Et; Distel et al, 2009) were used as well as Tg(ca8-ADV. E1b:TagRFP)bz4Tg, also named PC-FyntagRFP-T, as a control (Matsui et al, 2014).…”

Section: Animal Husbandrymentioning

confidence: 99%

“…A pB-Tol2 construct was used as a backbone into which a PC-specific enhancer (Matsui et al, 2014)/CMV basal promoter was inserted followed by FyntagRFP (a membrane-targeted red fluorescent protein tagRFP) fused to a T2A-peptide (myrCasp8ER T2 ; Chu et al, 2008), the chicken β-globin intron and SV40 polyA sequences (GIpA). This resulted in the final construct pBS_Tol2-SalI-PC:-EcoRI:FP-SpeI-T2A-myrCasp8ER T2 -XbaI-GIpA-NotI (Fig.…”

Section: Cloning Of Constructsmentioning

confidence: 99%

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Caspase-mediated apoptosis induction in zebrafish cerebellar Purkinje neurons

et al. 2016

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The zebrafish is a well-established model organism in which to study in vivo mechanisms of cell communication, differentiation and function. Existing cell ablation methods are either invasive or they rely on the cellular expression of prokaryotic enzymes and the use of antibiotic drugs as cell death-inducing compounds. We have recently established a novel inducible genetic cell ablation system based on tamoxifen-inducible Caspase 8 activity, thereby exploiting mechanisms of cell death intrinsic to most cell types. Here, we prove its suitability in vivo by monitoring the ablation of cerebellar Purkinje cells (PCs) in transgenic zebrafish that co-express the inducible caspase and a fluorescent reporter. Incubation of larvae in tamoxifen for 8 h activated endogenous Caspase 3 and cell death, whereas incubation for 16 h led to the near-complete loss of PCs by apoptosis. We observed synchronous cell death autonomous to the PC population and phagocytosing microglia in the cerebellum, reminiscent of developmental apoptosis in the forebrain. Thus, induction of apoptosis through targeted activation of caspase by tamoxifen (ATTAC TM ) further expands the repertoire of genetic tools for conditional interrogation of cellular functions.

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Section: Inducible Caspase Under Transient Transgenic Conditionsmentioning

confidence: 99%

Section: Inducible Caspase Under Transient Transgenic Conditionsmentioning

confidence: 99%

Section: Animal Husbandrymentioning

confidence: 99%

Section: Cloning Of Constructsmentioning

confidence: 99%

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Caspase-mediated apoptosis induction in zebrafish cerebellar Purkinje neurons

et al. 2016

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“…Due to the simple and efficient transgenesis techniques available for zebrafish however, genetically-encoded calcium indicators have also been extensively used for the investigation of visual processing in the tectum (Bianco & Engert, 2015;Hunter et al, 2013;Nikolaou et al, 2012;Romano et al, 2015). This is in addition to circuits in the spinal cord (de Vico Fallani et al, 2015;Warp et al, 2012), cerebellum (Matsui et al, 2014) and nucleus of the medial longitudinal fasciculus (nMLF) (Thiele et al, 2014).…”

Section: Optogenetic Dissection Of Zebrafish Neural Circuitsmentioning

confidence: 99%

Functional investigation of neural circuits in larval zebrafish

Thompson¹

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A fundamental goal of modern neuroscience is to understand the principles by which neural circuits within the brain lead to cognition and behaviour. Technological advances in the fields of microscopy and optogenetics have very recently allowed for the activity of all cells within a functional circuit to be observed and recorded, meaning that this goal is now within reach. Due to the physical and optical properties of neural tissue, these technologies remain most useful for the analysis of circuits that span very small volumes, meaning that small animal model organisms, with brains less than approximately 1 mm across, are most amenable to these types of analyses. The larval zebrafish model has emerged as an optimal balance between small size and the high complexity of a vertebrate brain. This thesis describes the development and application of several of these technological advances to examine neural circuits underlying perception and behaviour in the larval zebrafish.Due to the relative recency of zebrafish as a model for functional neuroscience, the genetic and experimental tools required for in depth circuit analysis have not been widely available. Thirteen transgenic zebrafish lines have been developed during this thesis for expressing optogenetic proteins throughout the brain, such as GCaMP5G for imaging neuronal calcium dynamics, and ChR2(ET/TC) and eNpHR3.0 for activating and silencing neuronal activity, respectively. These biological tools were developed in combination with optical techniques for probing neural circuits. This thesis describes the design and construction of a custom selective plane illumination microscope for the rapid imaging of neuronal activity across large areas of the zebrafish brain. The use of a spatial light modulator is also described for the targeted stimulation of ChR2(ET/TC) in the cerebellum of larval zebrafish. This experiment provided preliminary data for the functional investigation of connections between cerebellar cells and the optic tectum, as well as confirming the validity of this technique for analysis of other neuronal circuits in vivo.Since learning is one of the most interesting yet ill-defined processes controlled by the brain, this thesis aimed to use the tools developed above to examine the changes across neuronal circuit activity responsible for learning in larval zebrafish, specifically those in the cerebellum responsible for motor learning. A classical P ag e 2 conditioning assay was administered to 7-day-old zebrafish by pairing a non-startling tone with an aversive tail shock stimulus. This paradigm has been extensively used in other model organisms, however failed to produce a significant learning behaviour in the experiments performed for this thesis. More recent evidence suggests that zebrafish only begin to show learning from approximately one month of age.However, the data presented here indicate that zebrafish up to six weeks of age were still unable to undergo classical conditioning to pairings of auditory and electric stimuli. A number of is...

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Diversity of cellular physiology and morphology of Purkinje cells in the adult zebrafish cerebellum

Magnus

Xing

Zhang

et al. 2022

J of Comparative Neurology

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This study was designed to explore the functional circuitry of the adult zebrafish cerebellum, focusing on its Purkinje cells and using whole-cell patch recordings and single cell labeling in slice preparations. Following physiological characterizations, the recorded single cells were labeled for morphological identification. It was found that the zebrafish Purkinje cells are surprisingly diverse. Based on their physiology and morphology, they can be classified into at least three subtypes: Type I, a narrow spike cell, which fires only narrow Na + spikes (<3 ms in duration), and has a single primary dendrite with an arbor restricted to the distal molecular layer; Type II, a broad spike cell, which fires broad Ca 2+ spikes (5-7 ms in duration) and has a primary dendrite with limited branching in the inner molecular layer and then further radiates throughout the molecular layer; and Type III, a very broad spike cell, which fires very broad Ca 2+ spikes (≥10 ms in duration) and has a dense proximal dendritic arbor that is either restricted to the inner molecular layer (Type IIIa), or radiates throughout the entire molecular layer (Type IIIb). The graded paired-pulse facilitation of these Purkinje cells' responses to parallel fiber activations and the all-or-none, paired-pulse depression of climbing fiber activation are largely similar to those reported for mammals. The labeled axon terminals of these Purkinje cells end locally, as reported for larval zebrafish.The present study provides evidence that the corresponding functional circuitry and information processing differ from what has been well-established in the mammalian cerebellum.

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Functional regionalization of the teleost cerebellum analyzed in vivo

Cited by 84 publications

References 43 publications

Caspase-mediated apoptosis induction in zebrafish cerebellar Purkinje neurons

Caspase-mediated apoptosis induction in zebrafish cerebellar Purkinje neurons

Functional investigation of neural circuits in larval zebrafish

Diversity of cellular physiology and morphology of Purkinje cells in the adult zebrafish cerebellum

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