Superconcentration of light: circumventing the classical limit to achievable irradiance

Roth, Stephan; Sheppard, Colin J. R.; Heintzmann, Rainer

doi:10.1364/ol.41.002109

Cited by 33 publications

(22 citation statements)

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“…To localize actin cappers in stereocilia, we used conventional confocal microscopy or, for increased resolution, image-scanning microscopy with an Airyscan detector ( Müller and Enderlein, 2010 ; Sheppard et al, 2013 ; Roth et al, 2016 ). We used an antibody against CAPZB2 that has been validated with knockdown ( Sinnar et al, 2014 ) and peptide-blocking experiments (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We performed image-scanning microscopy ( Müller and Enderlein, 2010 ; Sheppard et al, 2013 ; Roth et al, 2016 ) using a 63×, 1.4 NA Plan-Apochromat objective or 25× 0.8 NA Plan-Neofluar ( Fig. 6, A and B ) on a Zeiss Elyra PS.1, equipped with an Airyscan detector; images were processed using ZEN 2012 (black edition, 64-bit software; Zeiss) version 8.0 with default settings.…”

Section: Methodsmentioning

confidence: 99%

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Heterodimeric capping protein is required for stereocilia length and width regulation

Avenarius

Krey

Dumont

et al. 2017

Journal of Cell Biology

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Heterodimeric capping protein is required for stereocilia length and width regulation

Avenarius

Krey

Dumont

et al. 2017

Journal of Cell Biology

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“…We occasionally detected GFP-ELMOD1 expression in cochlear hair cells. To provide higher-resolution localization of GFP-ELMOD1 there, we used image-scanning microscopy with an Airyscan detector (Müller and Enderlein, 2010;Sheppard et al, 2013;Roth et al, 2016). Some anti-GFP signal was detected in the stereocilia membrane of outer hair cells, but the anti-GFP signal was much brighter in tubular structures in the cells' somas ( Fig.…”

Section: Localization Of Elmod1mentioning

confidence: 99%

ELMOD1 Stimulates ARF6-GTP Hydrolysis to Stabilize Apical Structures in Developing Vestibular Hair Cells

et al. 2017

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Sensory hair cells require control of physical properties of their apical plasma membranes for normal development and function. Members of the ADP-ribosylation factor (ARF) small GTPase family regulate membrane trafficking and cytoskeletal assembly in many cells. We identified ELMO domain-containing protein 1 (ELMOD1), a guanine nucleoside triphosphatase activating protein (GAP) for ARF6, as the most highly enriched ARF regulator in hair cells. To characterize ELMOD1 control of trafficking, we analyzed mice of both sexes from a strain lacking functional ELMOD1 [roundabout ()]. In mice, cuticular plates of utricle hair cells initially formed normally, then degenerated after postnatal day 5; large numbers of vesicles invaded the compromised cuticular plate. Hair bundles initially developed normally, but the cell's apical membrane lifted away from the cuticular plate, and stereocilia elongated and fused. Membrane trafficking in type I hair cells, measured by FM1-43 dye labeling, was altered in mice. Consistent with the proposed GAP role for ELMOD1, the ARF6 GTP/GDP ratio was significantly elevated in utricles compared with controls, and the level of ARF6-GTP was correlated with the severity of the phenotype. These results suggest that conversion of ARF6 to its GDP-bound form is necessary for final stabilization of the hair bundle. Assembly of the mechanically sensitive hair bundle of sensory hair cells requires growth and reorganization of apical actin and membrane structures. Hair bundles and apical membranes in mice with mutations in the gene degenerate after formation, suggesting that the ELMOD1 protein stabilizes these structures. We show that ELMOD1 is a GTPase-activating protein in hair cells for the small GTP-binding protein ARF6, known to participate in actin assembly and membrane trafficking. We propose that conversion of ARF6 into the GDP-bound form in the apical domain of hair cells is essential for stabilizing apical actin structures like the hair bundle and ensuring that the apical membrane forms appropriately around the stereocilia.

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“…We occasionally detected GFP-ELMOD1 expression in cochlear hair cells. To provide higher-resolution localization of GFP-ELMOD1 there, we used imagescanning microscopy with an Airyscan detector (Müller and Enderlein, 2010;Sheppard et al, 2013;Roth et al, 2016). Some anti-GFP signal was detected in the stereocilia membrane of outer hair cells, but the anti-GFP signal was much brighter in tubular structures in the cells' somas ( Fig.…”

Section: Localization Of Elmod1mentioning

confidence: 99%

ELMOD1 stimulates ARF6-GTP hydrolysis to stabilize apical structures in developing vestibular hair cells

Krey

Dumont

Wilmarth

et al. 2017

Preprint

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Abbreviated title: ELMOD1 controls apical membranes through ARF6 11 figures, 0 tables, 0 multimedia, and 0 3D models Abstract, 191 words (250 maximum); Significance Statement, 110 words (120 maximum); Introduction, 618 words (650 maximum); Discussion, 1571 words (1500 maximum) Conflict of Interest: NoneAcknowledgements: We thank David Corey for sharing transcriptomics data prior to publication. PGBG was supported by NIH grants R01 DC002368 and P30 DC005983. We received support from the following core facilities: mass spectrometry from the OHSU Proteomics Shared Resource (partial support from NIH core grant P30 EY010572), confocal microscopy from the OHSU Advanced Light Microscopy Core @ The Jungers Center. Hybridoma cells for JLA20 (deposited by J. J.-C. Lin) and DSHB-GFP-4C9were obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242.All rights reserved. No reuse allowed without permission.(which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint . http://dx.doi.org/10.1101/189621 doi: bioRxiv preprint first posted online Sep. 15, 2017; 2 Abstract Sensory hair cells require control of physical properties of their apical plasma membranes for normal development and function. Members of the ARF small GTPase family regulate membrane trafficking and cytoskeletal assembly in many cells. We identified ELMOD1, a guanine nucleoside triphosphatase activating protein (GAP) for ARF6, as the most highly enriched ARF regulator in hair cells. To characterize ELMOD1 control of trafficking, we used a mouse strain lacking functional ELMOD1 (roundabout; rda). In rda/rda mice, cuticular plates of utricle hair cells initially formed normally, then degenerated after postnatal day 5 (P5); large numbers of vesicles invaded the compromised cuticular plate. Hair bundles initially developed normally, but the cell's apical membrane lifted away from the cuticular plate, and stereocilia elongated and fused. Membrane trafficking in type I hair cells, measured by FM1-43 dye labeling, was altered in rda/rda mice. Consistent with the proposed GAP role for ELMOD1, the ARF6 GTP/GDP ratio was significantly elevated in rda/rda utricles as compared to controls, and the level of ARF6-GTP was correlated with the severity of the rda/rda phenotype. These results suggest that conversion of ARF6 to its GDP-bound form is necessary for final stabilization of the hair bundle. Significance StatementAssembly of the mechanically sensitive hair bundle of sensory hair cells requires growth and reorganization of apical actin and membrane structures. Hair bundles and apical membranes in mice with mutations in the Elmod1 gene degenerate after formation, suggesting that the ELMOD1 protein stabilizes these structures. We show that ELMOD1 is a GTPase-activating protein in hair cells for the small GTPbinding protein ARF6, known to participate in act...

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Superconcentration of light: circumventing the classical limit to achievable irradiance

Cited by 33 publications

References 19 publications

Heterodimeric capping protein is required for stereocilia length and width regulation

Heterodimeric capping protein is required for stereocilia length and width regulation

ELMOD1 Stimulates ARF6-GTP Hydrolysis to Stabilize Apical Structures in Developing Vestibular Hair Cells

ELMOD1 stimulates ARF6-GTP hydrolysis to stabilize apical structures in developing vestibular hair cells

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