L-type voltage-gated calcium channel agonists mitigate hearing loss and modify ribbon synapse morphology in the zebrafish model of usher syndrome type 1

Alaa Koleilat; Joseph A. Dugdale; Trace A. Christenson; Jeffrey L. Bellah; Aaron M. Lambert; Mark A. Masino; Stephen C. Ekker; Lisa A. Schimmenti

doi:10.1242/DMM.043885

L-type voltage-gated calcium channel agonists mitigate hearing loss and modify ribbon synapse morphology in the zebrafish model of usher syndrome type 1

Alaa Koleilat, Joseph A. Dugdale, Trace A. Christenson, Jeffrey L. Bellah, Aaron M. Lambert, Mark A. Masino, Stephen C. Ekker, Lisa A. Schimmenti

Biochemistry and Molecular Biology

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

The mariner (myo7aa⁻/⁻) mutant is a zebrafish model for Usher syndrome type 1 (USH1). To further characterize hair cell synaptic elements in myo7aa⁻/⁻ mutants, we focused on the ribbon synapse and evaluated ultrastructure, number and distribution of immunolabeled ribbons, and postsynaptic densities. By transmission electron microscopy, we determined that myo7aa⁻/− zebrafish have fewer glutamatergic vesicles tethered to ribbon synapses, yet maintain a comparable ribbon area. In myo7aa⁻/− hair cells, immunolocalization of Ctbp2 showed fewer ribbon-containing cells in total and an altered distribution of Ctbp2 puncta compared to wild-type hair cells. myo7aa⁻/− mutants have fewer postsynaptic densities - as assessed by MAGUK immunolabeling - compared to wild-type zebrafish. We quantified the circular swimming behavior of myo7aa⁻/− mutant fish and measured a greater turning angle (absolute smooth orientation). It has previously been shown that L-type voltage-gated calcium channels are necessary for ribbon localization and occurrence of postsynaptic density; thus, we hypothesized and observed that L-type voltage-gated calcium channel agonists change behavioral and synaptic phenotypes in myo7aa⁻/− mutants in a drug-specific manner. Our results indicate that treatment with L-type voltage-gated calcium channel agonists alter hair cell synaptic elements and improve behavioral phenotypes of myo7aa⁻/− mutants. Our data support that L-type voltage-gated calcium channel agonists induce morphological changes at the ribbon synapse - in both the number of tethered vesicles and regarding the distribution of Ctbp2 puncta - shift swimming behavior and improve acoustic startle response.

Original language	English (US)
Article number	dmm043885
Journal	DMM Disease Models and Mechanisms
Volume	13
Issue number	11
DOIs	https://doi.org/10.1242/DMM.043885
State	Published - Nov 27 2020

Keywords

Hair cell
Hearing loss
Myo7aa
Ribbon synapse
Zebrafish

ASJC Scopus subject areas

Neuroscience (miscellaneous)
Medicine (miscellaneous)
Immunology and Microbiology (miscellaneous)
Biochemistry, Genetics and Molecular Biology(all)

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10.1242/DMM.043885

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Koleilat, A., Dugdale, J. A., Christenson, T. A., Bellah, J. L., Lambert, A. M., Masino, M. A., Ekker, S. C., & Schimmenti, L. A. (2020). L-type voltage-gated calcium channel agonists mitigate hearing loss and modify ribbon synapse morphology in the zebrafish model of usher syndrome type 1. DMM Disease Models and Mechanisms, 13(11), Article dmm043885. https://doi.org/10.1242/DMM.043885

@article{66243d7df2014236b4fd2895db0f397a,

title = "L-type voltage-gated calcium channel agonists mitigate hearing loss and modify ribbon synapse morphology in the zebrafish model of usher syndrome type 1",

abstract = "The mariner (myo7aa−/−) mutant is a zebrafish model for Usher syndrome type 1 (USH1). To further characterize hair cell synaptic elements in myo7aa−/− mutants, we focused on the ribbon synapse and evaluated ultrastructure, number and distribution of immunolabeled ribbons, and postsynaptic densities. By transmission electron microscopy, we determined that myo7aa−/− zebrafish have fewer glutamatergic vesicles tethered to ribbon synapses, yet maintain a comparable ribbon area. In myo7aa−/− hair cells, immunolocalization of Ctbp2 showed fewer ribbon-containing cells in total and an altered distribution of Ctbp2 puncta compared to wild-type hair cells. myo7aa−/− mutants have fewer postsynaptic densities - as assessed by MAGUK immunolabeling - compared to wild-type zebrafish. We quantified the circular swimming behavior of myo7aa−/− mutant fish and measured a greater turning angle (absolute smooth orientation). It has previously been shown that L-type voltage-gated calcium channels are necessary for ribbon localization and occurrence of postsynaptic density; thus, we hypothesized and observed that L-type voltage-gated calcium channel agonists change behavioral and synaptic phenotypes in myo7aa−/− mutants in a drug-specific manner. Our results indicate that treatment with L-type voltage-gated calcium channel agonists alter hair cell synaptic elements and improve behavioral phenotypes of myo7aa−/− mutants. Our data support that L-type voltage-gated calcium channel agonists induce morphological changes at the ribbon synapse - in both the number of tethered vesicles and regarding the distribution of Ctbp2 puncta - shift swimming behavior and improve acoustic startle response.",

keywords = "Hair cell, Hearing loss, Myo7aa, Ribbon synapse, Zebrafish",

author = "Alaa Koleilat and Dugdale, {Joseph A.} and Christenson, {Trace A.} and Bellah, {Jeffrey L.} and Lambert, {Aaron M.} and Masino, {Mark A.} and Ekker, {Stephen C.} and Schimmenti, {Lisa A.}",

note = "Funding Information: We thank Dr Teresa Nicolson for providing us with the tc320b allele of mariner. We are grateful to Mark Curry and Patrick Jochim in the Mayo Clinic Media Services. We also thank Dr Gabriel Martinez Galvez and Dr Timothy Wiggins for help with Adobe Illustrator, MATLAB and the behavior assessment. We are also grateful to Dr Lavinia Sheets for her helpful discussions. Last, we thank the staff members in the zebrafish core facilities at Mayo Clinic and the University of Minnesota, Twin Cities. A.K. was supported by the National Center for Advancing Translational Sciences (NCATS; grant number: UL1 TR000135), a component of the National Institutes of Health (NIH). M.A.M was supported by the NIH (grant number: R01NS094176). L.A.S. was supported by funding from the Mayo Clinic Center for Individualized Medicine, Philanthropic support from the Nelson Family Fund and from the Mayo Clinic Department of Otorhinolaryngology. Funding Information: A.K. was supported by the National Center for Advancing Translational Sciences (NCATS; grant number: UL1 TR000135), a component of the National Institutes of Health (NIH). M.A.M was supported by the NIH (grant number: R01NS094176). L.A.S. was supported by funding from the Mayo Clinic Center for Individualized Medicine, Philanthropic support from the Nelson Family Fund and from the Mayo Clinic Department of Otorhinolaryngology. Publisher Copyright: {\textcopyright} 2020 Company of Biologists Ltd. All rights reserved.",

year = "2020",

month = nov,

day = "27",

doi = "10.1242/DMM.043885",

language = "English (US)",

volume = "13",

journal = "DMM Disease Models and Mechanisms",

issn = "1754-8403",

publisher = "Company of Biologists Ltd",

number = "11",

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TY - JOUR

T1 - L-type voltage-gated calcium channel agonists mitigate hearing loss and modify ribbon synapse morphology in the zebrafish model of usher syndrome type 1

AU - Koleilat, Alaa

AU - Dugdale, Joseph A.

AU - Christenson, Trace A.

AU - Bellah, Jeffrey L.

AU - Lambert, Aaron M.

AU - Masino, Mark A.

AU - Ekker, Stephen C.

AU - Schimmenti, Lisa A.

N1 - Funding Information: We thank Dr Teresa Nicolson for providing us with the tc320b allele of mariner. We are grateful to Mark Curry and Patrick Jochim in the Mayo Clinic Media Services. We also thank Dr Gabriel Martinez Galvez and Dr Timothy Wiggins for help with Adobe Illustrator, MATLAB and the behavior assessment. We are also grateful to Dr Lavinia Sheets for her helpful discussions. Last, we thank the staff members in the zebrafish core facilities at Mayo Clinic and the University of Minnesota, Twin Cities. A.K. was supported by the National Center for Advancing Translational Sciences (NCATS; grant number: UL1 TR000135), a component of the National Institutes of Health (NIH). M.A.M was supported by the NIH (grant number: R01NS094176). L.A.S. was supported by funding from the Mayo Clinic Center for Individualized Medicine, Philanthropic support from the Nelson Family Fund and from the Mayo Clinic Department of Otorhinolaryngology. Funding Information: A.K. was supported by the National Center for Advancing Translational Sciences (NCATS; grant number: UL1 TR000135), a component of the National Institutes of Health (NIH). M.A.M was supported by the NIH (grant number: R01NS094176). L.A.S. was supported by funding from the Mayo Clinic Center for Individualized Medicine, Philanthropic support from the Nelson Family Fund and from the Mayo Clinic Department of Otorhinolaryngology. Publisher Copyright: © 2020 Company of Biologists Ltd. All rights reserved.

PY - 2020/11/27

Y1 - 2020/11/27

N2 - The mariner (myo7aa−/−) mutant is a zebrafish model for Usher syndrome type 1 (USH1). To further characterize hair cell synaptic elements in myo7aa−/− mutants, we focused on the ribbon synapse and evaluated ultrastructure, number and distribution of immunolabeled ribbons, and postsynaptic densities. By transmission electron microscopy, we determined that myo7aa−/− zebrafish have fewer glutamatergic vesicles tethered to ribbon synapses, yet maintain a comparable ribbon area. In myo7aa−/− hair cells, immunolocalization of Ctbp2 showed fewer ribbon-containing cells in total and an altered distribution of Ctbp2 puncta compared to wild-type hair cells. myo7aa−/− mutants have fewer postsynaptic densities - as assessed by MAGUK immunolabeling - compared to wild-type zebrafish. We quantified the circular swimming behavior of myo7aa−/− mutant fish and measured a greater turning angle (absolute smooth orientation). It has previously been shown that L-type voltage-gated calcium channels are necessary for ribbon localization and occurrence of postsynaptic density; thus, we hypothesized and observed that L-type voltage-gated calcium channel agonists change behavioral and synaptic phenotypes in myo7aa−/− mutants in a drug-specific manner. Our results indicate that treatment with L-type voltage-gated calcium channel agonists alter hair cell synaptic elements and improve behavioral phenotypes of myo7aa−/− mutants. Our data support that L-type voltage-gated calcium channel agonists induce morphological changes at the ribbon synapse - in both the number of tethered vesicles and regarding the distribution of Ctbp2 puncta - shift swimming behavior and improve acoustic startle response.

AB - The mariner (myo7aa−/−) mutant is a zebrafish model for Usher syndrome type 1 (USH1). To further characterize hair cell synaptic elements in myo7aa−/− mutants, we focused on the ribbon synapse and evaluated ultrastructure, number and distribution of immunolabeled ribbons, and postsynaptic densities. By transmission electron microscopy, we determined that myo7aa−/− zebrafish have fewer glutamatergic vesicles tethered to ribbon synapses, yet maintain a comparable ribbon area. In myo7aa−/− hair cells, immunolocalization of Ctbp2 showed fewer ribbon-containing cells in total and an altered distribution of Ctbp2 puncta compared to wild-type hair cells. myo7aa−/− mutants have fewer postsynaptic densities - as assessed by MAGUK immunolabeling - compared to wild-type zebrafish. We quantified the circular swimming behavior of myo7aa−/− mutant fish and measured a greater turning angle (absolute smooth orientation). It has previously been shown that L-type voltage-gated calcium channels are necessary for ribbon localization and occurrence of postsynaptic density; thus, we hypothesized and observed that L-type voltage-gated calcium channel agonists change behavioral and synaptic phenotypes in myo7aa−/− mutants in a drug-specific manner. Our results indicate that treatment with L-type voltage-gated calcium channel agonists alter hair cell synaptic elements and improve behavioral phenotypes of myo7aa−/− mutants. Our data support that L-type voltage-gated calcium channel agonists induce morphological changes at the ribbon synapse - in both the number of tethered vesicles and regarding the distribution of Ctbp2 puncta - shift swimming behavior and improve acoustic startle response.

KW - Hair cell

KW - Hearing loss

KW - Myo7aa

KW - Ribbon synapse

KW - Zebrafish

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U2 - 10.1242/DMM.043885

DO - 10.1242/DMM.043885

M3 - Article

C2 - 33361086

AN - SCOPUS:85099076301

SN - 1754-8403

VL - 13

JO - DMM Disease Models and Mechanisms

JF - DMM Disease Models and Mechanisms

IS - 11

M1 - dmm043885

ER -

L-type voltage-gated calcium channel agonists mitigate hearing loss and modify ribbon synapse morphology in the zebrafish model of usher syndrome type 1

Abstract

Keywords

ASJC Scopus subject areas

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