328 results on '"Rubel EW"'
Search Results
2. BFGF AND PHOTORECEPTOR REGENERATION
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STEINBERG, RH, FARBMAN, AI, CALOF, AL, REH, TA, RAYMOND, PA, POWERS, MK, PUJOL, R, RUBEL, EW, LEWIS, J, MARGOLIS, FL, and JORGENSEN, JM
- Published
- 1991
3. In vitro analysis of optimal stimuli for phase-locking and time-delayed modulation of firing in avian nucleus laminaris neurons
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Reyes, AD, primary, Rubel, EW, additional, and Spain, WJ, additional
- Published
- 1996
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4. Cell cycle progression in gentamicin-damaged avian cochleas
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Bhave, SA, primary, Stone, JS, additional, Rubel, EW, additional, and Coltrera, MD, additional
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- 1995
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5. Glutamatergic inhibition of voltage-operated calcium channels in the avian cochlear nucleus
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Lachica, EA, primary, Rubsamen, R, additional, Zirpel, L, additional, and Rubel, EW, additional
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- 1995
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6. Activation of a metabotropic glutamate receptor increases intracellular calcium concentrations in neurons of the avian cochlear nucleus
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Zirpel, L, primary, Lachica, EA, additional, and Rubel, EW, additional
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- 1995
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7. The astrocytic response to afferent activity blockade in chick nucleus magnocellularis is independent of synaptic activation, age, and neuronal survival
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Canady, KS, primary, Hyson, RL, additional, and Rubel, EW, additional
- Published
- 1994
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8. Membrane properties underlying the firing of neurons in the avian cochlear nucleus
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Reyes, AD, primary, Rubel, EW, additional, and Spain, WJ, additional
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- 1994
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9. A biphasic change in ribosomal conformation during transneuronal degeneration is altered by inhibition of mitochondrial, but not cytoplasmic protein synthesis
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Garden, GA, primary, Canady, KS, additional, Lurie, DI, additional, Bothwell, M, additional, and Rubel, EW, additional
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- 1994
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10. Identification of hair cell progenitors and intermitotic migration of their nuclei in the normal and regenerating avian inner ear
- Author
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Tsue, TT, primary, Watling, DL, additional, Weisleder, P, additional, Coltrera, MD, additional, and Rubel, EW, additional
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- 1994
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11. A circuit for coding interaural time differences in the chick brainstem
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Overholt, EM, primary, Rubel, EW, additional, and Hyson, RL, additional
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- 1992
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12. Aberrant projection induced by otocyst removal maintains normal tonotopic organization in the chick cochlear nucleus
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Lippe, WR, primary, Fuhrmann, DS, additional, Yang, W, additional, and Rubel, EW, additional
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- 1992
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13. Rapid and reversible astrocytic reaction to afferent activity blockade in chick cochlear nucleus
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Canady, KS, primary and Rubel, EW, additional
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- 1992
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14. Mechanisms of hair cell death and protection.
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Cheng AG, Cunningham LL, Rubel EW, Cheng, Alan G, Cunningham, Lisa L, and Rubel, Edwin W
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- 2005
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15. The effect of perilymphatic tetrodotoxin on peripheral auditory system function.
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Arts HA, Norton SJ, and Rubel EW
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- 1999
16. Quantitative distribution of the glycine receptor in the auditory brain stem of the gerbil
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Sanes, DH, primary, Geary, WA, additional, Wooten, GF, additional, and Rubel, EW, additional
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- 1987
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17. Transneuronal regulation of protein synthesis in the brain-stem auditory system of the chick requires synaptic activation
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Hyson, RL, primary and Rubel, EW, additional
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- 1989
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18. Frequency-specific projections of individual neurons in chick brainstem auditory nuclei
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Young, SR, primary and Rubel, EW, additional
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- 1983
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19. Afferent influences on brain stem auditory nuclei of the chicken: presynaptic action potentials regulate protein synthesis in nucleus magnocellularis neurons
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Born, DE, primary and Rubel, EW, additional
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- 1988
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20. The ontogeny of inhibition and excitation in the gerbil lateral superior olive
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Sanes, DH, primary and Rubel, EW, additional
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- 1988
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21. Hair cell regeneration reveals central nervous system plasticity in the avian brain.
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Rubel EW, Woolley SMN, Goode CT, and Fuchs AF
- Published
- 2003
22. Multiple mechanisms of aminoglycoside ototoxicity are distinguished by subcellular localization of action.
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Wu P, Becker FB, Ogelman R, Camci ED, Linbo TH, Simon JA, Rubel EW, and Raible DW
- Abstract
Mechanosensory hair cells of the inner ears and lateral line of vertebrates display heightened vulnerability to environmental insult, with damage resulting in hearing and balance disorders. An important example is hair cell loss due to exposure to toxic agents including therapeutic drugs such as the aminoglycoside antibiotics such as neomycin and gentamicin and antineoplastic agents. We describe two distinct cellular pathways for aminoglycoside-induced hair cell death in zebrafish lateral line hair cells. Neomycin exposure results in death from acute exposure with most cells dying within 1 hour of exposure. By contrast, exposure to gentamicin results primarily in delayed hair cell death, taking up to 24 hours for maximal effect. Washout experiments demonstrate that delayed death does not require continuous exposure, demonstrating two mechanisms where downstream responses differ in their timing. Acute damage is associated with mitochondrial calcium fluxes and can be alleviated by the mitochondrially-targeted antioxidant mitoTEMPO, while delayed death is independent of these factors. Conversely delayed death is associated with lysosomal accumulation and is reduced by altering endolysosomal function, while acute death is not sensitive to lysosomal manipulations. These experiments reveal the complexity of responses of hair cells to closely related compounds, suggesting that intervention focusing on early events rather than specific death pathways may be a successful therapeutic strategy.
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- 2024
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23. An in vivo Biomarker to Characterize Ototoxic Compounds and Novel Protective Therapeutics.
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Bellairs JA, Redila VA, Wu P, Tong L, Webster A, Simon JA, Rubel EW, and Raible DW
- Abstract
There are no approved therapeutics for the prevention of hearing loss and vestibular dysfunction from drugs like aminoglycoside antibiotics. While the mechanisms underlying aminoglycoside ototoxicity remain unresolved, there is considerable evidence that aminoglycosides enter inner ear mechanosensory hair cells through the mechanoelectrical transduction (MET) channel. Inhibition of MET-dependent uptake with small molecules or modified aminoglycosides is a promising otoprotective strategy. To better characterize mammalian ototoxicity and aid in the translation of emerging therapeutics, a biomarker is needed. In the present study we propose that neonatal mice systemically injected with the aminoglycosides G418 conjugated to Texas Red (G418-TR) can be used as a histologic biomarker to characterize in vivo aminoglycoside toxicity. We demonstrate that postnatal day 5 mice, like older mice with functional hearing, show uptake and retention of G418-TR in cochlear hair cells following systemic injection. When we compare G418-TR uptake in other tissues, we find that kidney proximal tubule cells show similar retention. Using ORC-13661, an investigational hearing protection drug, we demonstrate in vivo inhibition of aminoglycoside uptake in mammalian hair cells. This work establishes how systemically administered fluorescently labeled ototoxins in the neonatal mouse can reveal important details about ototoxic drugs and protective therapeutics., Competing Interests: JAS, EWR, and DWR are cofounders of Oricula Therapeutics, which has licensed patents covering ORC-13661 from the University of Washington. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Bellairs, Redila, Wu, Tong, Webster, Simon, Rubel and Raible.)
- Published
- 2022
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24. Chloroquine kills hair cells in zebrafish lateral line and murine cochlear cultures: Implications for ototoxicity.
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Davis SN, Wu P, Camci ED, Simon JA, Rubel EW, and Raible DW
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- Animals, Antiviral Agents toxicity, Cells, Cultured, Chloroquine toxicity, Hair Cells, Auditory cytology, Larva drug effects, Mice, Models, Animal, Ototoxicity, Zebrafish, Cell Survival drug effects, Chloroquine analogs & derivatives, Hair Cells, Auditory drug effects, Hydroxychloroquine toxicity, Lateral Line System drug effects
- Abstract
Hearing and balance deficits have been reported during and following treatment with the antimalarial drug chloroquine. However, experimental work examining the direct actions of chloroquine on mechanoreceptive hair cells in common experimental models is lacking. This study examines the effects of chloroquine on hair cells using two common experimental models: the zebrafish lateral line and neonatal mouse cochlear cultures. Zebrafish larvae were exposed to varying concentrations of chloroquine phosphate or hydroxychloroquine for 1 h or 24 h, and hair cells assessed by antibody staining. A significant, dose-dependent reduction in the number of surviving hair cells was seen across conditions for both exposure periods. Hydroxychloroquine showed similar toxicity. In mouse cochlear cultures, chloroquine damage was specific to outer hair cells in tissue from the cochlear basal turn, consistent with susceptibility to other ototoxic agents. These findings suggest a need for future studies employing hearing and balance monitoring during exposure to chloroquine and related compounds, particularly with interest in these compounds as therapeutics against viral infections including coronavirus., (Copyright © 2020 Elsevier B.V. All rights reserved.)
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- 2020
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25. ORC-13661 protects sensory hair cells from aminoglycoside and cisplatin ototoxicity.
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Kitcher SR, Kirkwood NK, Camci ED, Wu P, Gibson RM, Redila VA, Simon JA, Rubel EW, Raible DW, Richardson GP, and Kros CJ
- Subjects
- Amikacin toxicity, Aminoglycosides toxicity, Animals, Cell Culture Techniques, Cells, Cultured, Cisplatin toxicity, Disease Models, Animal, Dose-Response Relationship, Drug, Hair Cells, Auditory metabolism, Humans, Intravital Microscopy, Ion Channels antagonists & inhibitors, Ion Channels metabolism, Male, Mechanotransduction, Cellular drug effects, Mice, Ototoxicity etiology, Patch-Clamp Techniques, Protective Agents therapeutic use, Rats, Thiophenes therapeutic use, Time-Lapse Imaging, Urea pharmacology, Urea therapeutic use, Zebrafish, Anti-Bacterial Agents toxicity, Antineoplastic Agents toxicity, Hair Cells, Auditory drug effects, Ototoxicity prevention & control, Protective Agents pharmacology, Thiophenes pharmacology, Urea analogs & derivatives
- Abstract
Aminoglycoside (AG) antibiotics are widely used to prevent life-threatening infections, and cisplatin is used in the treatment of various cancers, but both are ototoxic and result in loss of sensory hair cells from the inner ear. ORC-13661 is a new drug that was derived from PROTO-1, a compound first identified as protective in a large-scale screen utilizing hair cells in the lateral line organs of zebrafish larvae. Here, we demonstrate, in zebrafish larvae and in mouse cochlear cultures, that ORC-13661 provides robust protection of hair cells against both ototoxins, the AGs and cisplatin. ORC-13661 also prevents both hearing loss in a dose-dependent manner in rats treated with amikacin and the loading of neomycin-Texas Red into lateral line hair cells. In addition, patch-clamp recordings in mouse cochlear cultures reveal that ORC-13661 is a high-affinity permeant blocker of the mechanoelectrical transducer (MET) channel in outer hair cells, suggesting that it may reduce the toxicity of AGs by directly competing for entry at the level of the MET channel and of cisplatin by a MET-dependent mechanism. ORC-13661 is therefore a promising and versatile protectant that reversibly blocks the hair cell MET channel and operates across multiple species and toxins.
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- 2019
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26. De novo sequencing and initial annotation of the Mongolian gerbil (Meriones unguiculatus) genome.
- Author
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Zorio DAR, Monsma S, Sanes DH, Golding NL, Rubel EW, and Wang Y
- Subjects
- Animals, Base Sequence, Male, Molecular Sequence Annotation, Genome, Gerbillinae genetics, Sequence Analysis, DNA
- Abstract
The Mongolian gerbil (Meriones unguiculatus) is a member of the rodent family that displays several features not found in mice or rats, including sensory specializations and social patterns more similar to those in humans. These features have made gerbils a valuable animal for research studies of auditory and visual processing, brain development, learning and memory, and neurological disorders. Here, we report the whole gerbil annotated genome sequence, and identify important similarities and differences to the human and mouse genomes. We further analyze the chromosomal structure of eight genes with high relevance for controlling neural signaling and demonstrate a high degree of homology between these genes in mouse and gerbil. This homology increases the likelihood that individual genes can be rapidly identified in gerbil and used for genetic manipulations. The availability of the gerbil genome provides a foundation for advancing our knowledge towards understanding evolution, behavior and neural function in mammals. ACCESSION NUMBER: The Whole Genome Shotgun sequence data from this project has been deposited at DDBJ/ENA/GenBank under the accession NHTI00000000. The version described in this paper is version NHTI01000000. The fragment reads, and mate pair reads have been deposited in the Sequence Read Archive under BioSample accession SAMN06897401., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2019
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27. The role of retrograde intraflagellar transport genes in aminoglycoside-induced hair cell death.
- Author
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Stawicki TM, Linbo T, Hernandez L, Parkinson L, Bellefeuille D, Rubel EW, and Raible DW
- Abstract
Sensory hair cells are susceptible to numerous insults, including certain therapeutic medications like aminoglycoside antibiotics, and hearing and balance disorders are often a dose-limiting side effect of these medications. We show that mutations in multiple genes in both the retrograde intraflagellar transport (IFT) motor and adaptor complexes lead to resistance to aminoglycoside-induced hair cell death. These mutations also lead to defects in the entry of both aminoglycosides and the vital dye FM1-43 into hair cells, both processes that depend on hair cell mechanotransduction activity. However, the trafficking of proteins important for mechanotransduction activity is not altered by these mutations. Our data suggest that both retrograde IFT motor and adaptor complex genes are playing a role in aminoglycoside toxicity through affecting aminoglycoside uptake into hair cells., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2019. Published by The Company of Biologists Ltd.)
- Published
- 2019
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28. Noise-Induced Hypersensitization of the Acoustic Startle Response in Larval Zebrafish.
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Bhandiwad AA, Raible DW, Rubel EW, and Sisneros JA
- Subjects
- Animals, Behavior, Animal, Electric Stimulation, Larva physiology, Noise, Quinoxalines, Valine analogs & derivatives, alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid, Receptors, AMPA metabolism, Reflex, Startle, Zebrafish physiology
- Abstract
Overexposure to loud noise is known to lead to deficits in auditory sensitivity and perception. We studied the effects of noise exposure on sensorimotor behaviors of larval (5-7 days post-fertilization) zebrafish (Danio rerio), particularly the auditory-evoked startle response and hearing sensitivity to acoustic startle stimuli. We observed a temporary 10-15 dB decrease in startle response threshold after 18 h of flat-spectrum noise exposure at 20 dB re·1 ms
-2 . Larval zebrafish also exhibited decreased habituation to startle-inducing stimuli following noise exposure. The noise-induced sensitization was not due to changes in absolute hearing thresholds, but was specific to the auditory-evoked escape responses. The observed noise-induced sensitization was disrupted by AMPA receptor blockade using DNQX, but not NMDA receptor blockade. Together, these experiments suggest a complex effect of noise exposure on the neural circuits mediating auditory-evoked behaviors in larval zebrafish.- Published
- 2018
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29. Phenotypic Optimization of Urea-Thiophene Carboxamides To Yield Potent, Well Tolerated, and Orally Active Protective Agents against Aminoglycoside-Induced Hearing Loss.
- Author
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Chowdhury S, Owens KN, Herr RJ, Jiang Q, Chen X, Johnson G, Groppi VE, Raible DW, Rubel EW, and Simon JA
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- Administration, Oral, Animals, Drug Evaluation, Preclinical, Rats, Structure-Activity Relationship, Thiophenes administration & dosage, Thiophenes adverse effects, Zebrafish, Aminoglycosides adverse effects, Hearing Loss chemically induced, Hearing Loss prevention & control, Safety, Thiophenes chemistry, Thiophenes pharmacology, Urea chemistry
- Abstract
Hearing loss is a major public health concern with no pharmaceutical intervention for hearing protection or restoration. Using zebrafish neuromast hair cells, a robust model for mammalian auditory and vestibular hair cells, we identified a urea-thiophene carboxamide, 1 (ORC-001), as protective against aminoglycoside antibiotic (AGA)-induced hair cell death. The 50% protection (HC
50 ) concentration conferred by 1 is 3.2 μM with protection against 200 μM neomycin approaching 100%. Compound 1 was sufficiently safe and drug-like to validate otoprotection in an in vivo rat hearing loss model. We explored the structure-activity relationship (SAR) of this compound series to improve otoprotective potency, improve pharmacokinetic properties and eliminate off-target activity. We present the optimization of 1 to yield 90 (ORC-13661). Compound 90 protects mechanosensory hair cells with HC50 of 120 nM and demonstrates 100% protection in the zebrafish assay and superior physiochemical, pharmacokinetic, and toxicologic properties, as well as complete in vivo protection in rats.- Published
- 2018
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30. Proteomic analyses of nucleus laminaris identified candidate targets of the fragile X mental retardation protein.
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Sakano H, Zorio DAR, Wang X, Ting YS, Noble WS, MacCoss MJ, Rubel EW, and Wang Y
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- Animals, Blotting, Western, Brain Stem cytology, Chickens, Chromatography, Liquid, Dendrites metabolism, Dermoscopy, Electrophoresis, Escherichia coli, Immunohistochemistry, Laser Capture Microdissection, Microscopy, Confocal, Proteomics, Recombinant Proteins metabolism, Tandem Mass Spectrometry, rhoC GTP-Binding Protein metabolism, Avian Proteins metabolism, Brain Stem metabolism, Fragile X Mental Retardation Protein metabolism, Proteome
- Abstract
The avian nucleus laminaris (NL) is a brainstem nucleus necessary for binaural processing, analogous in structure and function to the mammalian medial superior olive. In chickens (Gallus gallus), NL is a well-studied model system for activity-dependent neural plasticity. Its neurons have bipolar extension of dendrites, which receive segregated inputs from two ears and display rapid and compartment-specific reorganization in response to unilateral changes in auditory input. More recently, fragile X mental retardation protein (FMRP), an RNA-binding protein that regulates local protein translation, has been shown to be enriched in NL dendrites, suggesting its potential role in the structural dynamics of these dendrites. To explore the molecular role of FMRP in this nucleus, we performed proteomic analysis of NL, using micro laser capture and liquid chromatography tandem mass spectrometry. We identified 657 proteins, greatly represented in pathways involved in mitochondria, translation and metabolism, consistent with high levels of activity of NL neurons. Of these, 94 are potential FMRP targets, by comparative analysis with previously proposed FMRP targets in mammals. These proteins are enriched in pathways involved in cellular growth, cellular trafficking and transmembrane transport. Immunocytochemistry verified the dendritic localization of several proteins in NL. Furthermore, we confirmed the direct interaction of FMRP with one candidate, RhoC, by in vitro RNA binding assays. In summary, we provide a database of highly expressed proteins in NL and in particular a list of potential FMRP targets, with the goal of facilitating molecular characterization of FMRP signaling in future studies., (© 2017 Wiley Periodicals, Inc.)
- Published
- 2017
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31. Cellular distribution of the fragile X mental retardation protein in the mouse brain.
- Author
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Zorio DA, Jackson CM, Liu Y, Rubel EW, and Wang Y
- Subjects
- Animals, Blotting, Western, Fragile X Syndrome metabolism, Gene Expression Profiling, Immunohistochemistry, Mice, Mice, Knockout, Transcriptome, Brain metabolism, Fragile X Mental Retardation Protein biosynthesis
- Abstract
The fragile X mental retardation protein (FMRP) plays an important role in normal brain development. Absence of FMRP results in abnormal neuronal morphologies in a selected manner throughout the brain, leading to intellectual deficits and sensory dysfunction in the fragile X syndrome (FXS). Despite FMRP importance for proper brain function, its overall expression pattern in the mammalian brain at the resolution of individual neuronal cell groups is not known. In this study we used FMR1 knockout and isogenic wildtype mice to systematically map the distribution of FMRP expression in the entire mouse brain. Using immunocytochemistry and cellular quantification analyses, we identified a large number of prominent cell groups expressing high levels of FMRP at the subcortical levels, in particular sensory and motor neurons in the brainstem and thalamus. In contrast, many cell groups in the midbrain and hypothalamus exhibit low FMRP levels. More important, we describe differential patterns of FMRP distribution in both cortical and subcortical brain regions. Almost all major brain areas contain high and low levels of FMRP cell groups adjacent to each other or between layers of the same cortical areas. These differential patterns indicate that FMRP expression appears to be specific to individual neuronal cell groups instead of being associated with all neurons in distinct brain regions, as previously considered. Taken together, these findings support the notion of FMRP differential neuronal regulation and strongly implicate the contribution of fundamental sensory and motor processing at subcortical levels to FXS pathology. J. Comp. Neurol. 525:818-849, 2017. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)
- Published
- 2017
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32. Fluorescent aminoglycosides reveal intracellular trafficking routes in mechanosensory hair cells.
- Author
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Hailey DW, Esterberg R, Linbo TH, Rubel EW, and Raible DW
- Subjects
- Animals, Biological Transport, Active drug effects, Cell Death drug effects, Cell Death genetics, Zebrafish genetics, Aminoglycosides pharmacokinetics, Aminoglycosides toxicity, Fluorescent Dyes pharmacokinetics, Fluorescent Dyes toxicity, Hair Cells, Vestibular metabolism, Zebrafish metabolism
- Abstract
Aminoglycosides (AGs) are broad-spectrum antibiotics that are associated with kidney damage, balance disorders, and permanent hearing loss. This damage occurs primarily by killing of proximal tubule kidney cells and mechanosensory hair cells, though the mechanisms underlying cell death are not clear. Imaging molecules of interest in living cells can elucidate how molecules enter cells, traverse intracellular compartments, and interact with sites of activity. Here, we have imaged fluorescently labeled AGs in live zebrafish mechanosensory hair cells. We determined that AGs enter hair cells via both nonendocytic and endocytic pathways. Both routes deliver AGs from the extracellular space to lysosomes, and structural differences between AGs alter the efficiency of this delivery. AGs with slower delivery to lysosomes were immediately toxic to hair cells, and impeding lysosome delivery increased AG-induced death. Therefore, pro-death cascades induced at early time points of AG exposure do not appear to derive from the lysosome. Our findings help clarify how AGs induce hair cell death and reveal properties that predict toxicity. Establishing signatures for AG toxicity may enable more efficient evaluation of AG treatment paradigms and structural modifications to reduce hair cell damage. Further, this work demonstrates how following fluorescently labeled drugs at high resolution in living cells can reveal important details about how drugs of interest behave., Competing Interests: The authors have declared that no conflict of interest exists.
- Published
- 2017
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33. Maintenance of neuronal size gradient in MNTB requires sound-evoked activity.
- Author
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Weatherstone JH, Kopp-Scheinpflug C, Pilati N, Wang Y, Forsythe ID, Rubel EW, and Tempel BL
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- 2-Amino-5-phosphonovalerate pharmacology, Animals, Deafness genetics, Diphtheria Toxin pharmacology, Evoked Potentials, Auditory genetics, Excitatory Amino Acid Antagonists pharmacology, Excitatory Postsynaptic Potentials drug effects, Excitatory Postsynaptic Potentials genetics, Homeodomain Proteins genetics, Homeodomain Proteins metabolism, Mice, Mice, Inbred CBA, Mice, Transgenic, Microtubule-Associated Proteins metabolism, Mutation genetics, Neurons physiology, Plasma Membrane Calcium-Transporting ATPases genetics, Plasma Membrane Calcium-Transporting ATPases metabolism, Presynaptic Terminals physiology, Sodium Channel Blockers pharmacology, Tetrodotoxin pharmacology, Transcription Factor Brn-3C genetics, Transcription Factor Brn-3C metabolism, Cochlear Nucleus pathology, Deafness pathology, Deafness physiopathology, Evoked Potentials, Auditory physiology, Neurons pathology, Sound
- Abstract
The medial nucleus of the trapezoid body (MNTB) is an important source of inhibition during the computation of sound location. It transmits fast and precisely timed action potentials at high frequencies; this requires an efficient calcium clearance mechanism, in which plasma membrane calcium ATPase 2 (PMCA2) is a key component. Deafwaddler ( dfw
2J ) mutant mice have a null mutation in PMCA2 causing deafness in homozygotes ( dfw2J / dfw2J ) and high-frequency hearing loss in heterozygotes (+/ dfw2J ). Despite the deafness phenotype, no significant differences in MNTB volume or cell number were observed in dfw2J homozygous mutants, suggesting that PMCA2 is not required for MNTB neuron survival. The MNTB tonotopic axis encodes high to low sound frequencies across the medial to lateral dimension. We discovered a cell size gradient along this axis: lateral neuronal somata are significantly larger than medially located somata. This size gradient is decreased in +/ dfw2J and absent in dfw2J / dfw2J The lack of acoustically driven input suggests that sound-evoked activity is required for maintenance of the cell size gradient. This hypothesis was corroborated by selective elimination of auditory hair cell activity with either hair cell elimination in Pou4f3 DTR mice or inner ear tetrodotoxin (TTX) treatment. The change in soma size was reversible and recovered within 7 days of TTX treatment, suggesting that regulation of the gradient is dependent on synaptic activity and that these changes are plastic rather than permanent. NEW & NOTEWORTHY Neurons of the medial nucleus of the trapezoid body (MNTB) act as fast-spiking inhibitory interneurons within the auditory brain stem. The MNTB is topographically organized, with low sound frequencies encoded laterally and high frequencies medially. We discovered a cell size gradient along this axis: lateral neurons are larger than medial neurons. The absence of this gradient in deaf mice lacking plasma membrane calcium ATPase 2 suggests an activity-dependent, calcium-mediated mechanism that controls neuronal soma size., (Copyright © 2017 the American Physiological Society.)- Published
- 2017
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34. Glial Cell Contributions to Auditory Brainstem Development.
- Author
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Cramer KS and Rubel EW
- Subjects
- Animals, Auditory Pathways growth & development, Brain Stem growth & development, Auditory Pathways physiology, Brain Stem physiology, Neuroglia physiology
- Abstract
Glial cells, previously thought to have generally supporting roles in the central nervous system, are emerging as essential contributors to multiple aspects of neuronal circuit function and development. This review focuses on the contributions of glial cells to the development of auditory pathways in the brainstem. These pathways display specialized synapses and an unusually high degree of precision in circuitry that enables sound source localization. The development of these pathways thus requires highly coordinated molecular and cellular mechanisms. Several classes of glial cells, including astrocytes, oligodendrocytes and microglia, have now been explored in these circuits in both avian and mammalian brainstems. Distinct populations of astrocytes are found over the course of auditory brainstem maturation. Early appearing astrocytes are associated with spatial compartments in the avian auditory brainstem. Factors from late appearing astrocytes promote synaptogenesis and dendritic maturation, and astrocytes remain integral parts of specialized auditory synapses. Oligodendrocytes play a unique role in both birds and mammals in highly regulated myelination essential for proper timing to decipher interaural cues. Microglia arise early in brainstem development and may contribute to maturation of auditory pathways. Together these studies demonstrate the importance of non-neuronal cells in the assembly of specialized auditory brainstem circuits.
- Published
- 2016
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35. Mitochondrial calcium uptake underlies ROS generation during aminoglycoside-induced hair cell death.
- Author
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Esterberg R, Linbo T, Pickett SB, Wu P, Ou HC, Rubel EW, and Raible DW
- Subjects
- Aminoglycosides pharmacology, Animals, Cytoplasm metabolism, Disease Models, Animal, Lateral Line System, Oxidation-Reduction, Oxygen chemistry, Transgenes, Zebrafish, Aminoglycosides adverse effects, Calcium metabolism, Cell Death drug effects, Hair Cells, Auditory drug effects, Mitochondria metabolism, Reactive Oxygen Species metabolism
- Abstract
Exposure to aminoglycoside antibiotics can lead to the generation of toxic levels of reactive oxygen species (ROS) within mechanosensory hair cells of the inner ear that have been implicated in hearing and balance disorders. Better understanding of the origin of aminoglycoside-induced ROS could focus the development of therapies aimed at preventing this event. In this work, we used the zebrafish lateral line system to monitor the dynamic behavior of mitochondrial and cytoplasmic oxidation occurring within the same dying hair cell following exposure to aminoglycosides. The increased oxidation observed in both mitochondria and cytoplasm of dying hair cells was highly correlated with mitochondrial calcium uptake. Application of the mitochondrial uniporter inhibitor Ru360 reduced mitochondrial and cytoplasmic oxidation, suggesting that mitochondrial calcium drives ROS generation during aminoglycoside-induced hair cell death. Furthermore, targeting mitochondria with free radical scavengers conferred superior protection against aminoglycoside exposure compared with identical, untargeted scavengers. Our findings suggest that targeted therapies aimed at preventing mitochondrial oxidation have therapeutic potential to ameliorate the toxic effects of aminoglycoside exposure.
- Published
- 2016
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36. Cilia-Associated Genes Play Differing Roles in Aminoglycoside-Induced Hair Cell Death in Zebrafish.
- Author
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Stawicki TM, Hernandez L, Esterberg R, Linbo T, Owens KN, Shah AN, Thapa N, Roberts B, Moens CB, Rubel EW, and Raible DW
- Subjects
- Animals, Cell Death, Cilia metabolism, Cilia ultrastructure, Cytoplasmic Dyneins genetics, Cytoplasmic Dyneins metabolism, Gene Expression, Hair Cells, Auditory cytology, Hair Cells, Auditory metabolism, Mechanotransduction, Cellular, Microtubule-Associated Proteins genetics, Microtubule-Associated Proteins metabolism, Vesicular Transport Proteins genetics, Vesicular Transport Proteins metabolism, Zebrafish, Zebrafish Proteins genetics, Zebrafish Proteins metabolism, Aminoglycosides toxicity, Cilia drug effects, Drug Tolerance genetics, Hair Cells, Auditory drug effects, Mutation
- Abstract
Hair cells possess a single primary cilium, called the kinocilium, early in development. While the kinocilium is lost in auditory hair cells of most species it is maintained in vestibular hair cells. It has generally been believed that the primary role of the kinocilium and cilia-associated genes in hair cells is in the establishment of the polarity of actin-based stereocilia, the hair cell mechanotransduction apparatus. Through genetic screening and testing of candidate genes in zebrafish (Danio rerio) we have found that mutations in multiple cilia genes implicated in intraflagellar transport (dync2h1, wdr35, ift88, and traf3ip), and the ciliary transition zone (cc2d2a, mks1, and cep290) lead to resistance to aminoglycoside-induced hair cell death. These genes appear to have differing roles in hair cells, as mutations in intraflagellar transport genes, but not transition zone genes, lead to defects in kinocilia formation and processes dependent upon hair cell mechanotransduction activity. These mutants highlight a novel role of cilia-associated genes in hair cells, and provide powerful tools for further study., (Copyright © 2016 Stawicki et al.)
- Published
- 2016
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37. Innervation regulates synaptic ribbons in lateral line mechanosensory hair cells.
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Suli A, Pujol R, Cunningham DE, Hailey DW, Prendergast A, Rubel EW, and Raible DW
- Subjects
- Animals, Hair Cells, Auditory, Inner ultrastructure, Lateral Line System ultrastructure, Membranes metabolism, Mutation genetics, Synapses ultrastructure, Synaptic Vesicles metabolism, Synaptic Vesicles ultrastructure, Zebrafish, Hair Cells, Auditory, Inner metabolism, Lateral Line System innervation, Lateral Line System metabolism, Mechanotransduction, Cellular, Synapses metabolism
- Abstract
Failure to form proper synapses in mechanosensory hair cells, the sensory cells responsible for hearing and balance, leads to deafness and balance disorders. Ribbons are electron-dense structures that tether synaptic vesicles to the presynaptic zone of mechanosensory hair cells where they are juxtaposed with the post-synaptic endings of afferent fibers. They are initially formed throughout the cytoplasm, and, as cells mature, ribbons translocate to the basolateral membrane of hair cells to form functional synapses. We have examined the effect of post-synaptic elements on ribbon formation and maintenance in the zebrafish lateral line system by observing mutants that lack hair cell innervation, wild-type larvae whose nerves have been transected and ribbons in regenerating hair cells. Our results demonstrate that innervation is not required for initial ribbon formation but suggest that it is crucial for regulating the number, size and localization of ribbons in maturing hair cells, and for ribbon maintenance at the mature synapse., (© 2016. Published by The Company of Biologists Ltd.)
- Published
- 2016
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38. Systematic and differential myelination of axon collaterals in the mammalian auditory brainstem.
- Author
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Seidl AH and Rubel EW
- Subjects
- Animals, Auditory Pathways growth & development, Brain Stem growth & development, Cell Size, Gerbillinae, Imaging, Three-Dimensional, Lysine analogs & derivatives, Microscopy, Confocal, Auditory Pathways cytology, Axons, Brain Stem cytology, Myelin Sheath
- Abstract
A brainstem circuit for encoding the spatial location of sounds involves neurons in the cochlear nucleus that project to medial superior olivary (MSO) neurons on both sides of the brain via a single bifurcating axon. Neurons in MSO act as coincidence detectors, responding optimally when signals from the two ears arrive within a few microseconds. To achieve this, transmission of signals along the contralateral collateral must be faster than transmission of the same signals along the ipsilateral collateral. We demonstrate that this is achieved by differential regulation of myelination and axon caliber along the ipsilateral and contralateral branches of single axons; ipsilateral axon branches have shorter internode lengths and smaller caliber than contralateral branches. The myelination difference is established prior to the onset of hearing. We conclude that this differential myelination and axon caliber requires local interactions between axon collaterals and surrounding oligodendrocytes on the two sides of the brainstem., (© 2015 Wiley Periodicals, Inc.)
- Published
- 2016
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39. Fractalkine Signaling Regulates Macrophage Recruitment into the Cochlea and Promotes the Survival of Spiral Ganglion Neurons after Selective Hair Cell Lesion.
- Author
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Kaur T, Zamani D, Tong L, Rubel EW, Ohlemiller KK, Hirose K, and Warchol ME
- Subjects
- Animals, Cell Survival physiology, Cochlea cytology, Cochlea physiology, Female, Humans, Male, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Mice, Transgenic, Chemokine CX3CL1 physiology, Hair Cells, Auditory physiology, Macrophages physiology, Signal Transduction physiology, Spiral Ganglion cytology, Spiral Ganglion physiology
- Abstract
Macrophages are recruited into the cochlea in response to injury caused by acoustic trauma or ototoxicity, but the nature of the interaction between macrophages and the sensory structures of the inner ear remains unclear. The present study examined the role of fractalkine signaling in regulating the injury-evoked behavior of macrophages following the selective ablation of cochlear hair cells. We used a novel transgenic mouse model in which the human diphtheria toxin receptor (huDTR) is selectively expressed under the control of Pou4f3, a hair cell-specific transcription factor. Administration of diphtheria toxin (DT) to these mice resulted in nearly complete ablation of cochlear hair cells, with no evident pathology among supporting cells, spiral ganglion neurons, or cells of the cochlear lateral wall. Hair cell death led to an increase in macrophages associated with the sensory epithelium of the cochlea. Their numbers peaked at 14 days after DT and then declined at later survival times. Increased macrophages were also observed within the spiral ganglion, but their numbers remained elevated for (at least) 56 d after DT. To investigate the role of fractalkine signaling in macrophage recruitment, we crossed huDTR mice to a mouse line that lacks expression of the fractalkine receptor (CX3CR1). Disruption of fractalkine signaling reduced macrophage recruitment into both the sensory epithelium and spiral ganglion and also resulted in diminished survival of spiral ganglion neurons after hair cell death. Our results suggest a fractalkine-mediated interaction between macrophages and the neurons of the cochlea., Significance Statement: It is known that damage to the inner ear leads to recruitment of inflammatory cells (macrophages), but the chemical signals that initiate this recruitment and the functions of macrophages in the damaged ear are unclear. Here we show that fractalkine signaling regulates macrophage recruitment into the cochlea and also promotes the survival of cochlear afferents after selective hair cell lesion. Because these afferent neurons carry sound information from the cochlea to the auditory brainstem, their survival is a key determinant of the success of cochlear prosthetics. Our data suggest that fractalkine signaling in the cochlea is neuroprotective, and reveal a previously uncharacterized interaction between cells of the cochlea and the innate immune system., (Copyright © 2015 the authors 0270-6474/15/3515050-12$15.00/0.)
- Published
- 2015
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40. Selective deletion of cochlear hair cells causes rapid age-dependent changes in spiral ganglion and cochlear nucleus neurons.
- Author
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Tong L, Strong MK, Kaur T, Juiz JM, Oesterle EC, Hume C, Warchol ME, Palmiter RD, and Rubel EW
- Subjects
- Animals, Cell Death, Cochlear Nucleus cytology, Cochlear Nucleus physiology, Diphtheria Toxin pharmacology, Hair Cells, Auditory drug effects, Hair Cells, Auditory metabolism, Hearing, Heparin-binding EGF-like Growth Factor genetics, Heparin-binding EGF-like Growth Factor metabolism, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Spiral Ganglion cytology, Spiral Ganglion physiology, Cochlear Nucleus growth & development, Hair Cells, Auditory cytology, Spiral Ganglion growth & development
- Abstract
During nervous system development, critical periods are usually defined as early periods during which manipulations dramatically change neuronal structure or function, whereas the same manipulations in mature animals have little or no effect on the same property. Neurons in the ventral cochlear nucleus (CN) are dependent on excitatory afferent input for survival during a critical period of development. Cochlear removal in young mammals and birds results in rapid death of target neurons in the CN. Cochlear removal in older animals results in little or no neuron death. However, the extent to which hair-cell-specific afferent activity prevents neuronal death in the neonatal brain is unknown. We further explore this phenomenon using a new mouse model that allows temporal control of cochlear hair cell deletion. Hair cells express the human diphtheria toxin (DT) receptor behind the Pou4f3 promoter. Injections of DT resulted in nearly complete loss of organ of Corti hair cells within 1 week of injection regardless of the age of injection. Injection of DT did not influence surrounding supporting cells directly in the sensory epithelium or spiral ganglion neurons (SGNs). Loss of hair cells in neonates resulted in rapid and profound neuronal loss in the ventral CN, but not when hair cells were eliminated at a more mature age. In addition, normal survival of SGNs was dependent on hair cell integrity early in development and less so in mature animals. This defines a previously undocumented critical period for SGN survival., (Copyright © 2015 the authors 0270-6474/15/357878-14$15.00/0.)
- Published
- 2015
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41. Macrophage recruitment and epithelial repair following hair cell injury in the mouse utricle.
- Author
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Kaur T, Hirose K, Rubel EW, and Warchol ME
- Abstract
The sensory organs of the inner ear possess resident populations of macrophages, but the function of those cells is poorly understood. In many tissues, macrophages participate in the removal of cellular debris after injury and can also promote tissue repair. The present study examined injury-evoked macrophage activity in the mouse utricle. Experiments used transgenic mice in which the gene for the human diphtheria toxin receptor (huDTR) was inserted under regulation of the Pou4f3 promoter. Hair cells in such mice can be selectively lesioned by systemic treatment with diphtheria toxin (DT). In order to visualize macrophages, Pou4f3-huDTR mice were crossed with a second transgenic line, in which one or both copies of the gene for the fractalkine receptor CX3CR1 were replaced with a gene for GFP. Such mice expressed GFP in all macrophages, and mice that were CX3CR1(GFP/GFP) lacked the necessary receptor for fractalkine signaling. Treatment with DT resulted in the death of ∼70% of utricular hair cells within 7 days, which was accompanied by increased numbers of macrophages within the utricular sensory epithelium. Many of these macrophages appeared to be actively engulfing hair cell debris, indicating that macrophages participate in the process of 'corpse removal' in the mammalian vestibular organs. However, we observed no apparent differences in injury-evoked macrophage numbers in the utricles of CX3CR1(+/GFP) mice vs. CX3CR1(GFP/GFP) mice, suggesting that fractalkine signaling is not necessary for macrophage recruitment in these sensory organs. Finally, we found that repair of sensory epithelia at short times after DT-induced hair cell lesions was mediated by relatively thin cables of F-actin. After 56 days recovery, however, all cell-cell junctions were characterized by very thick actin cables.
- Published
- 2015
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42. Identification of small molecule inhibitors of cisplatin-induced hair cell death: results of a 10,000 compound screen in the zebrafish lateral line.
- Author
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Thomas AJ, Wu P, Raible DW, Rubel EW, Simon JA, and Ou HC
- Subjects
- Animals, Dose-Response Relationship, Drug, Drug Evaluation, Preclinical, Hearing Loss chemically induced, Zebrafish, Antineoplastic Agents toxicity, Cell Death drug effects, Cisplatin toxicity, Hair Cells, Auditory drug effects, Hearing Loss prevention & control, Lateral Line System drug effects, Neuroprotective Agents pharmacology
- Abstract
Hypothesis: The zebrafish lateral line can be used to identify small molecules that protect against cisplatin-induced hair cell death., Background: Cisplatin is a commonly used chemotherapeutic agent, which causes hearing loss by damaging hair cells of the inner ear. There are currently no FDA-approved pharmacologic strategies for preventing this side effect. The zebrafish lateral line has been used successfully in the past to study hair cell death and protection., Methods: In this study, we used the zebrafish lateral line to screen a library of 10,000 small molecules for protection against cisplatin-induced hair cell death. Dose-response relationships for identified protectants were determined by quantifying hair cell protection. The effect of each protectant on uptake of a fluorescent cisplatin analog was also quantified., Results: From this screen, we identified 2 compounds exhibiting dose-dependent protection: cisplatin hair cell protectant 1 and 2 (CHCP1 and 2). CHCP1 reduced the uptake of a fluorescent cisplatin analog, suggesting its protective effects may be due to decreased cisplatin uptake. CHCP2 did not affect uptake, which suggests an intracellular mechanism of action. Evaluation of analogs of CHCP2 revealed 3 additional compounds that significantly reduced cisplatin-induced hair cell death, although none exceed the effectiveness or potency of the parent compound., Conclusion: The zebrafish lateral line was used to identify 2 small molecules that protected against cisplatin-induced hair cell death.
- Published
- 2015
- Full Text
- View/download PDF
43. Seasonal plasticity of precise spike timing in the avian auditory system.
- Author
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Caras ML, Sen K, Rubel EW, and Brenowitz EA
- Subjects
- Animals, Auditory Pathways cytology, Auditory Pathways metabolism, Auditory Pathways physiology, Female, Gonadal Steroid Hormones blood, Male, Neurons physiology, Prosencephalon cytology, Prosencephalon metabolism, Sparrows, Evoked Potentials, Auditory, Neuronal Plasticity, Photoperiod, Prosencephalon physiology, Seasons
- Abstract
Vertebrate audition is a dynamic process, capable of exhibiting both short- and long-term adaptations to varying listening conditions. Precise spike timing has long been known to play an important role in auditory encoding, but its role in sensory plasticity remains largely unexplored. We addressed this issue in Gambel's white-crowned sparrow (Zonotrichia leucophrys gambelii), a songbird that shows pronounced seasonal fluctuations in circulating levels of sex-steroid hormones, which are known to be potent neuromodulators of auditory function. We recorded extracellular single-unit activity in the auditory forebrain of males and females under different breeding conditions and used a computational approach to explore two potential strategies for the neural discrimination of sound level: one based on spike counts and one based on spike timing reliability. We report that breeding condition has robust sex-specific effects on spike timing. Specifically, in females, breeding condition increases the proportion of cells that rely solely on spike timing information and increases the temporal resolution required for optimal intensity encoding. Furthermore, in a functionally distinct subset of cells that are particularly well suited for amplitude encoding, female breeding condition enhances spike timing-based discrimination accuracy. No effects of breeding condition were observed in males. Our results suggest that high-resolution temporal discharge patterns may provide a plastic neural substrate for sensory coding., (Copyright © 2015 the authors 0270-6474/15/353431-15$15.00/0.)
- Published
- 2015
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44. Using the zebrafish lateral line to uncover novel mechanisms of action and prevention in drug-induced hair cell death.
- Author
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Stawicki TM, Esterberg R, Hailey DW, Raible DW, and Rubel EW
- Abstract
The majority of hearing loss and balance disorders are caused by the permanent loss of mechanosensory hair cells of the inner ear. Identification of genes and compounds that modulate susceptibility to hair cell death is frequently confounded by the difficulties of assaying for such complex phenomena in mammalian models. The zebrafish has emerged as a powerful animal model for genetic and chemical screening in many contexts. Several characteristics of the zebrafish, such as its small size and external location of mechanosensory hair cells within the lateral line sensory organ, uniquely position it as an ideal model organism for the study of hair cell toxicity. We have used this model to screen for genes and compounds that affect hair cell survival during ototoxin exposure and have identified agents that would not be expected to play a role in this process based on a priori knowledge of their function. The identification of such agents yields better understanding of hair cell death and holds promise to stem hearing loss and balance disorders in the human population.
- Published
- 2015
- Full Text
- View/download PDF
45. ER-mitochondrial calcium flow underlies vulnerability of mechanosensory hair cells to damage.
- Author
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Esterberg R, Hailey DW, Rubel EW, and Raible DW
- Subjects
- Aminoglycosides pharmacology, Animals, Animals, Genetically Modified, Cell Death drug effects, Chelating Agents pharmacology, Cytoplasm drug effects, Cytoplasm metabolism, Dose-Response Relationship, Drug, Egtazic Acid pharmacology, Embryo, Nonmammalian, Endoplasmic Reticulum drug effects, Endoplasmic Reticulum genetics, Larva, Lateral Line System anatomy & histology, Mechanoreceptors drug effects, Membrane Potential, Mitochondrial drug effects, Mitochondria drug effects, Myosin Heavy Chains genetics, Time Factors, Transcription Factor Brn-3C genetics, Zebrafish, Calcium metabolism, Endoplasmic Reticulum metabolism, Mechanoreceptors metabolism, Mitochondria metabolism
- Abstract
Mechanosensory hair cells are vulnerable to environmental insult, resulting in hearing and balance disorders. We demonstrate that directional compartmental flow of intracellular Ca(2+) underlies death in zebrafish lateral line hair cells after exposure to aminoglycoside antibiotics, a well characterized hair cell toxin. Ca(2+) is mobilized from the ER and transferred to mitochondria via IP3 channels with little cytoplasmic leakage. Pharmacological agents that shunt ER-derived Ca(2+) directly to cytoplasm mitigate toxicity, indicating that high cytoplasmic Ca(2+) levels alone are not cytotoxic. Inhibition of the mitochondrial transition pore sensitizes hair cells to the toxic effects of aminoglycosides, contrasting with current models of excitotoxicity. Hair cells display efficient ER-mitochondrial Ca(2+) flow, suggesting that tight coupling of these organelles drives mitochondrial activity under physiological conditions at the cost of increased susceptibility to toxins., (Copyright © 2014 the authors 0270-6474/14/349703-17$15.00/0.)
- Published
- 2014
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46. The zebrafish merovingian mutant reveals a role for pH regulation in hair cell toxicity and function.
- Author
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Stawicki TM, Owens KN, Linbo T, Reinhart KE, Rubel EW, and Raible DW
- Subjects
- Acids metabolism, Amino Acid Sequence, Animals, Cisplatin toxicity, DNA-Binding Proteins chemistry, DNA-Binding Proteins metabolism, Extracellular Space drug effects, Extracellular Space metabolism, Hair Cells, Auditory, Inner drug effects, Hair Cells, Auditory, Inner enzymology, Hydrogen-Ion Concentration, Molecular Sequence Data, Mutation, Missense genetics, Neomycin toxicity, Proton-Translocating ATPases metabolism, Pyridinium Compounds metabolism, Quaternary Ammonium Compounds metabolism, Transcription Factors chemistry, Transcription Factors metabolism, Zebrafish Proteins chemistry, Zebrafish Proteins metabolism, DNA-Binding Proteins genetics, Hair Cells, Auditory, Inner metabolism, Hair Cells, Auditory, Inner pathology, Transcription Factors genetics, Zebrafish genetics, Zebrafish Proteins genetics
- Abstract
Control of the extracellular environment of inner ear hair cells by ionic transporters is crucial for hair cell function. In addition to inner ear hair cells, aquatic vertebrates have hair cells on the surface of their body in the lateral line system. The ionic environment of these cells also appears to be regulated, although the mechanisms of this regulation are less understood than those of the mammalian inner ear. We identified the merovingian mutant through genetic screening in zebrafish for genes involved in drug-induced hair cell death. Mutants show complete resistance to neomycin-induced hair cell death and partial resistance to cisplatin-induced hair cell death. This resistance is probably due to impaired drug uptake as a result of reduced mechanotransduction ability, suggesting that the mutants have defects in hair cell function independent of drug treatment. Through genetic mapping we found that merovingian mutants contain a mutation in the transcription factor gcm2. This gene is important for the production of ionocytes, which are cells crucial for whole body pH regulation in fish. We found that merovingian mutants showed an acidified extracellular environment in the vicinity of both inner ear and lateral line hair cells. We believe that this acidified extracellular environment is responsible for the defects seen in hair cells of merovingian mutants, and that these mutants would serve as a valuable model for further study of the role of pH in hair cell function., (© 2014. Published by The Company of Biologists Ltd.)
- Published
- 2014
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47. Intense and specialized dendritic localization of the fragile X mental retardation protein in binaural brainstem neurons: a comparative study in the alligator, chicken, gerbil, and human.
- Author
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Wang Y, Sakano H, Beebe K, Brown MR, de Laat R, Bothwell M, Kulesza RJ Jr, and Rubel EW
- Subjects
- Aged, Aged, 80 and over, Animals, Avian Proteins genetics, Avian Proteins metabolism, Female, Fragile X Mental Retardation Protein genetics, Humans, Male, Middle Aged, Rats, Sprague-Dawley metabolism, Reptilian Proteins genetics, Reptilian Proteins metabolism, Sequence Homology, Amino Acid, Species Specificity, Superior Olivary Complex metabolism, Alligators and Crocodiles metabolism, Brain Stem metabolism, Chickens metabolism, Dendrites metabolism, Fragile X Mental Retardation Protein metabolism, Gerbillinae metabolism
- Abstract
Neuronal dendrites are structurally and functionally dynamic in response to changes in afferent activity. The fragile X mental retardation protein (FMRP) is an mRNA binding protein that regulates activity-dependent protein synthesis and morphological dynamics of dendrites. Loss and abnormal expression of FMRP occur in fragile X syndrome (FXS) and some forms of autism spectrum disorders. To provide further understanding of how FMRP signaling regulates dendritic dynamics, we examined dendritic expression and localization of FMRP in the reptilian and avian nucleus laminaris (NL) and its mammalian analogue, the medial superior olive (MSO), in rodents and humans. NL/MSO neurons are specialized for temporal processing of low-frequency sounds for binaural hearing, which is impaired in FXS. Protein BLAST analyses first demonstrate that the FMRP amino acid sequences in the alligator and chicken are highly similar to human FMRP with identical mRNA-binding and phosphorylation sites, suggesting that FMRP functions similarly across vertebrates. Immunocytochemistry further reveals that NL/MSO neurons have very high levels of dendritic FMRP in low-frequency hearing vertebrates including alligator, chicken, gerbil, and human. Remarkably, dendritic FMRP in NL/MSO neurons often accumulates at branch points and enlarged distal tips, loci known to be critical for branch-specific dendritic arbor dynamics. These observations support an important role for FMRP in regulating dendritic properties of binaural neurons that are essential for low-frequency sound localization and auditory scene segregation, and support the relevance of studying this regulation in nonhuman vertebrates that use low frequencies in order to further understand human auditory processing disorders., (Copyright © 2013 Wiley Periodicals, Inc.)
- Published
- 2014
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- View/download PDF
48. Differential conduction velocity regulation in ipsilateral and contralateral collaterals innervating brainstem coincidence detector neurons.
- Author
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Seidl AH, Rubel EW, and Barría A
- Subjects
- Animals, Axons physiology, Chick Embryo, Electric Stimulation, Excitatory Amino Acid Antagonists pharmacology, Female, GABA Antagonists pharmacology, Imaging, Three-Dimensional, In Vitro Techniques, Male, Models, Neurological, Neural Conduction drug effects, Neurons drug effects, Patch-Clamp Techniques, Picrotoxin pharmacology, Quinoxalines pharmacology, Valine analogs & derivatives, Valine pharmacology, Brain Stem cytology, Functional Laterality physiology, Neural Conduction physiology, Neural Pathways physiology, Neurons physiology
- Abstract
Information processing in the brain relies on precise timing of signal propagation. The highly conserved neuronal network for computing spatial representations of acoustic signals resolves microsecond timing of sounds processed by the two ears. As such, it provides an excellent model for understanding how precise temporal regulation of neuronal signals is achieved and maintained. The well described avian and mammalian brainstem circuit for computation of interaural time differences is composed of monaural cells in the cochlear nucleus (CN; nucleus magnocellularis in birds) projecting to binaurally innervated coincidence detection neurons in the medial superior olivary nucleus (MSO) in mammals or nucleus laminaris (NL) in birds. Individual axons from CN neurons issue a single axon that bifurcates into an ipsilateral branch and a contralateral branch that innervate segregated dendritic regions of the MSO/NL coincidence detector neurons. We measured conduction velocities of the ipsilateral and contralateral branches of these bifurcating axon collaterals in the chicken by antidromic stimulation of two sites along each branch and whole-cell recordings in the parent neurons. At the end of each experiment, the individual CN neuron and its axon collaterals were filled with dye. We show that the two collaterals of a single axon adjust the conduction velocities individually to achieve the specific conduction velocities essential for precise temporal integration of information from the two ears, as required for sound localization. More generally, these results suggest that individual axonal segments in the CNS interact locally with surrounding neural structures to determine conduction velocity.
- Published
- 2014
- Full Text
- View/download PDF
49. Spontaneous hair cell regeneration in the neonatal mouse cochlea in vivo.
- Author
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Cox BC, Chai R, Lenoir A, Liu Z, Zhang L, Nguyen DH, Chalasani K, Steigelman KA, Fang J, Rubel EW, Cheng AG, and Zuo J
- Subjects
- Animals, Anion Transport Proteins metabolism, Hair Cells, Auditory ultrastructure, Mice, Microscopy, Electron, Scanning, Mitosis physiology, Sulfate Transporters, Animals, Newborn, Cell Transdifferentiation physiology, Hair Cells, Auditory physiology, Regeneration physiology
- Abstract
Loss of cochlear hair cells in mammals is currently believed to be permanent, resulting in hearing impairment that affects more than 10% of the population. Here, we developed two genetic strategies to ablate neonatal mouse cochlear hair cells in vivo. Both Pou4f3(DTR/+) and Atoh1-CreER™; ROSA26(DTA/+) alleles allowed selective and inducible hair cell ablation. After hair cell loss was induced at birth, we observed spontaneous regeneration of hair cells. Fate-mapping experiments demonstrated that neighboring supporting cells acquired a hair cell fate, which increased in a basal to apical gradient, averaging over 120 regenerated hair cells per cochlea. The normally mitotically quiescent supporting cells proliferated after hair cell ablation. Concurrent fate mapping and labeling with mitotic tracers showed that regenerated hair cells were derived by both mitotic regeneration and direct transdifferentiation. Over time, regenerated hair cells followed a similar pattern of maturation to normal hair cell development, including the expression of prestin, a terminal differentiation marker of outer hair cells, although many new hair cells eventually died. Hair cell regeneration did not occur when ablation was induced at one week of age. Our findings demonstrate that the neonatal mouse cochlea is capable of spontaneous hair cell regeneration after damage in vivo. Thus, future studies on the neonatal cochlea might shed light on the competence of supporting cells to regenerate hair cells and on the factors that promote the survival of newly regenerated hair cells.
- Published
- 2014
- Full Text
- View/download PDF
50. Bax, Bcl2, and p53 differentially regulate neomycin- and gentamicin-induced hair cell death in the zebrafish lateral line.
- Author
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Coffin AB, Rubel EW, and Raible DW
- Subjects
- Animals, Cell Death drug effects, Cell Death physiology, Female, Gene Expression Regulation, Developmental, Gentamicins toxicity, Hair Cells, Auditory physiology, Lateral Line System embryology, Male, Neomycin toxicity, Protein Synthesis Inhibitors toxicity, Proto-Oncogene Proteins c-bcl-2 metabolism, Tumor Suppressor Protein p53 metabolism, Zebrafish embryology, Zebrafish Proteins metabolism, bcl-2-Associated X Protein metabolism, Hair Cells, Auditory pathology, Lateral Line System pathology, Proto-Oncogene Proteins c-bcl-2 genetics, Tumor Suppressor Protein p53 genetics, Zebrafish genetics, Zebrafish Proteins genetics, bcl-2-Associated X Protein genetics
- Abstract
Sensorineural hearing loss is a normal consequence of aging and results from a variety of extrinsic challenges such as excessive noise exposure and certain therapeutic drugs, including the aminoglycoside antibiotics. The proximal cause of hearing loss is often death of inner ear hair cells. The signaling pathways necessary for hair cell death are not fully understood and may be specific for each type of insult. In the lateral line, the closely related aminoglycoside antibiotics neomycin and gentamicin appear to kill hair cells by activating a partially overlapping suite of cell death pathways. The lateral line is a system of hair cell-containing sense organs found on the head and body of aquatic vertebrates. In the present study, we use a combination of pharmacologic and genetic manipulations to assess the contributions of p53, Bax, and Bcl2 in the death of zebrafish lateral line hair cells. Bax inhibition significantly protects hair cells from neomycin but not from gentamicin toxicity. Conversely, transgenic overexpression of Bcl2 attenuates hair cell death due to gentamicin but not neomycin, suggesting a complex interplay of pro-death and pro-survival proteins in drug-treated hair cells. p53 inhibition protects hair cells from damage due to either aminoglycoside, with more robust protection seen against gentamicin. Further experiments evaluating p53 suggest that inhibition of mitochondrial-specific p53 activity confers significant hair cell protection from either aminoglycoside. These results suggest a role for mitochondrial p53 activity in promoting hair cell death due to aminoglycosides, likely upstream of Bax and Bcl2.
- Published
- 2013
- Full Text
- View/download PDF
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