An Assay for Systematically Quantifying the Vestibulo-Ocular Reflex to Assess Vestibular Function in Zebrafish Larvae

Sun, Peng; Zhang, Yingla; Zhao, Feng; Wu, Jianping; Pun, Sio Hang; Peng, Cheng; Du, Meide; Vai, Mang I; Liu, Dong; Chen, Fangyi

doi:10.3389/fncel.2018.00257

Cited by 12 publications

(16 citation statements)

References 33 publications

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“…At this stage of development, the larvae are free swimming and reflexive movements of the trunk muscles in zebrafish larvae are predicted to arise in response to vestibular cues. Rotation of the head does not activate the immature semicircular canals at this stage of development, but rather leads to excitation of vestibular hair cells in the anterior maculae (Beck et al, 2004;Mo et al, 2010;Sun et al, 2018).…”

Section: The Vsr In Zebrafish Larvaementioning

confidence: 92%

“…The construction of VSR test device was described in Sun et al (2018). We designed a new mounting chamber with a narrow slot to immobilize the head of larvae, which we inserted into a 5 mm wide slot on the chamber holder.…”

Section: Vsr Testmentioning

confidence: 99%

“…For our experiments, we elicited tail movements in headfixed larvae with a yaw stimulus in dark conditions and recorded responses using a device that was previously used to assess vestibular-induced eye movements in larvae (Sun et al, 2018). Briefly, the main components of the test device for infrared (IR) imaging include a rotary platform driven by a motor, an objective lens (10x), an IR light emitting diode (LED) to illuminate the larvae, an IR camera ( Figure 1A), and a small chamber unit for mounting the larvae that can be inserted into the chamber holder ( Figure 1B).…”

Section: The Vsr In Zebrafish Larvaementioning

confidence: 99%

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Temporal Vestibular Deficits in synaptojanin 1 (synj1) Mutants

Gao

Nicolson

2021

Front. Mol. Neurosci.

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The lipid phosphatase synaptojanin 1 (synj1) is required for the disassembly of clathrin coats on endocytic compartments. In neurons such activity is necessary for the recycling of endocytosed membrane into synaptic vesicles. Mutations in zebrafish synj1 have been shown to disrupt the activity of ribbon synapses in sensory hair cells. After prolonged mechanical stimulation of hair cells, both phase locking of afferent nerve activity and the recovery of spontaneous release of synaptic vesicles are diminished in synj1 mutants. Presumably as a behavioral consequence of these synaptic deficits, synj1 mutants are unable to maintain an upright posture. To probe vestibular function with respect to postural control in synj1 mutants, we developed a method for assessing the vestibulospinal reflex (VSR) in larvae. We elicited the VSR by rotating the head and recorded tail movements. As expected, the VSR is completely absent in pcdh15a and lhfpl5a mutants that lack inner ear function. Conversely, lhfpl5b mutants, which have a selective loss of function of the lateral line organ, have normal VSRs, suggesting that the hair cells of this organ do not contribute to this reflex. In contrast to mechanotransduction mutants, the synj1 mutant produces normal tail movements during the initial cycles of rotation of the head. Both the amplitude and temporal aspects of the response are unchanged. However, after several rotations, the VSR in synj1 mutants was strongly diminished or absent. Mutant synj1 larvae are able to recover, but the time required for the reappearance of the VSR after prolonged stimulation is dramatically increased in synj1 mutants. Collectively, the data demonstrate a behavioral correlate of the synaptic defects caused by the loss of synj1 function. Our results suggest that defects in synaptic vesicle recycling give rise to fatigue of ribbons synapses and possibly other synapses of the VS circuit, leading to the loss of postural control.

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Section: The Vsr In Zebrafish Larvaementioning

confidence: 92%

Section: Vsr Testmentioning

confidence: 99%

Section: The Vsr In Zebrafish Larvaementioning

confidence: 99%

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Temporal Vestibular Deficits in synaptojanin 1 (synj1) Mutants

Gao

Nicolson

2021

Front. Mol. Neurosci.

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“…The zebrafish larva was gently mounted in the larva-shaped chamber in a dorsal-up position with the tail glued by 5% methylcellulose and covered with a piece of glass coverslip on the chamber. After adding E3 embryo media in the head region, the chamber unit was then mounted on a device for quantifying linear vestibulo-ocular reflex (VOR) from Southern University of Science and Technology ( Sun et al, 2018 ). After aligning the larval eyes to the center of the infrared camera, the platform started to rotate back and forth around a horizontal axis at a speed of 30 rpm, and the VOR was recorded by the camera.…”

Section: Methodsmentioning

confidence: 99%

Claudin h Is Essential for Hair Cell Morphogenesis and Auditory Function in Zebrafish

Gong

Qian

et al. 2021

Front. Cell Dev. Biol.

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Hereditary hearing loss caused by defective hair cells is one of the most common congenital diseases, whose nosogenesis is still unclear because many of the causative genes remain unidentified. Claudins are one kind of transmembrane proteins that constitute the most important components of the tight junctions and paracellular barrier and play important roles in neurodevelopment. In this study, we investigated the function of claudin h in morphogenesis and auditory function of the hair cell in zebrafish. The results of in situ hybridization showed that claudin h was specifically localized in the otic vesicle and neuromasts in zebrafish embryos. The deficiency of claudin h caused significant reduction of otic vesicle size and loss of utricle otolith. Moreover, the startle response and vestibulo-ocular reflex experiments revealed that loss of claudin h led to serious hearing loss and vestibular dysfunction. Importantly, the confocal microscopy observation found that compared to the control zebrafish, the claudin h morphants and mutants displayed significantly reduced the number of cristae hair cells and shortened kinocilia. Besides, the deficiency of claudin h also caused the loss of hair cells in neuromasts which could be rescued by injecting claudin h mRNA into the mutant embryos at one cell stage. Furthermore, the immunohistochemistry experiments demonstrated remarkable apoptosis of hair cells in the neuromasts, which might contribute to the loss of hair cells number. Overall, these data indicated that claudin h is indispensable for the development of hair cells, vestibular function, and hearing ability of zebrafish.

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“…Vestibular head tilt response was measured as described previously (Sun et al, 2018). Briefly, individual zebrafish larva at 5 dpf was placed in a customized holder, where its tail was glued to immobilize the fish.…”

Section: Vestibular Head Tilt Response Measurementmentioning

confidence: 99%

Rbm24a Is Necessary for Hair Cell Development Through Regulating mRNA Stability in Zebrafish

Zhang

Wang

Yao

et al. 2020

Front. Cell Dev. Biol.

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Hair cells in the inner ear and lateral lines are mechanosensitive receptor cells whose development and function are tightly regulated. Several transcription factors as well as splicing factors have been identified to play important roles in hair cell development, whereas the role of RNA stability in this process is poorly understood. In the present work, we report that RNA-binding motif protein 24a (Rbm24a) is indispensable for hair cell development in zebrafish. Rbm24a expression is detected in the inner ear as well as lateral line neuromasts. Albeit rbm24a deficient zebrafish do not survive beyond 9 days post fertilization (dpf) due to effects outside of the inner ear, rbm24a deficiency does not affect the early development of inner ear except for delayed otolith formation and semicircular canal fusion. However, hair cell development is severely affected and hair bundle is disorganized in rbm24a mutants. As a result, the auditory and vestibular function of rbm24a mutants are compromised. RNAseq analyses identified several Rbm24a-target mRNAs that are directly bound by Rbm24a and are dysregulated in rbm24a mutants. Among the identified Rbm24a-target genes, lrrc23, dfna5b, and smpx are particularly interesting as their dysregulation might contribute to the inner ear phenotypes in rbm24a mutants. In conclusion, our data suggest that Rbm24a affects hair cell development in zebrafish through regulating mRNA stability.

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An Assay for Systematically Quantifying the Vestibulo-Ocular Reflex to Assess Vestibular Function in Zebrafish Larvae

Cited by 12 publications

References 33 publications

Temporal Vestibular Deficits in synaptojanin 1 (synj1) Mutants

Temporal Vestibular Deficits in synaptojanin 1 (synj1) Mutants

Claudin h Is Essential for Hair Cell Morphogenesis and Auditory Function in Zebrafish

Rbm24a Is Necessary for Hair Cell Development Through Regulating mRNA Stability in Zebrafish

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