1996
DOI: 10.1159/000259200
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Immunologically Defined Component of the Circumferential Ring Around the Cuticular Plate in Mammalian Hair Cells

Abstract: A monoclonal antibody (CAR) was raised by in vitro immunisation to a component of the circumferential actin ring that is associated with the apical junctions encircling the cuticular plates of mammalian hair cells. On western blots it bound a protein band at about 42 kD, equivalent to the normal location of actin, but it did not label the paracrystalline bundle of actin filaments in the stereocilia, the complex actin filament gel that forms the cuticular plate or the filamentous actin in the cell cortex. When … Show more

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Cited by 5 publications
(6 citation statements)
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References 24 publications
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“…We have raised numerous monoclonal antibodies to mammalian hair cells with in vitro immunization by using antigen prepared from guinea pig organs of Corti (Holley and Richardson, 1994;Holley and Nishida, 1995;Nishida and Holley, 1996). Many of the antibodies raised by using this method react with antigens in the cuticular plate, stereocilia, and numerous additional components of the reticular lamina.…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…We have raised numerous monoclonal antibodies to mammalian hair cells with in vitro immunization by using antigen prepared from guinea pig organs of Corti (Holley and Richardson, 1994;Holley and Nishida, 1995;Nishida and Holley, 1996). Many of the antibodies raised by using this method react with antigens in the cuticular plate, stereocilia, and numerous additional components of the reticular lamina.…”
Section: Methodsmentioning
confidence: 99%
“…The immunization method yields many antibodies to inner ear antigens, but few of those that are effective for immunofluorescence can also be used for immunoblotting. Two exceptions are antibodies against the lateral cisternae of guinea pig outer hair cells (Holley and Richardson, 1994) and the circumferential actin ring (Nishida and Holley, 1996).…”
Section: Possible Antigens For Cp1 Sc1 and Sp1-sp3mentioning
confidence: 99%
“…A blocking solution of 5% goat serum in PBS was used for 10 min before the antibody was added. The primary antibodies used for the characterization were as follows: anti-fimbrin antibody (rabbit polyclonal, dilution 1 : 25, a kind gift from Dr P. Matsudaira, Whitehead Institute for Biomedical Research, MIT, Cambridge, MA, USA); cuticular plate 1 (CP1) (dilution 1 : 100), stereocilia and pillar cells 1 (SP1) (dilution 1 : 100) and stereocilia 1 (SC1) (dilution 1 : 100), three mouse monoclonals raised against cochlear tissue (Nishida & Holley, 1996;Nishida et al, 1998); anti-myosin VIIa (rabbit polyclonal, dilution 1 : 100, a kind gift from Professor C. Petit, Institut Pasteur, Paris, France); anti-myosin VI (rabbit polyclonal, dilution 1 : 60, a gift of Dr T. Hasson, University of California at San Diego, USA); anti-espin antibody (rabbit polyclonal, dilution 1 : 7.6, a gift from Dr J. Bartles, Northwestern University, Chicago, USA) and anti-p27 kip1 (Abcam, Cambridge, UK). As a control for non-specific labelling, primary antibodies were replaced with either mouse or rabbit non-reactive immunoglobulins.…”
Section: Immunocytochemistrymentioning
confidence: 99%
“…To analyse the actin cytoskeleton more closely we used three monoclonal antibodies that recognize epitopes expressed by differentiating hair cells (Nishida & Holley, 1996;Nishida et al, 1998). During inner ear development in the mouse CP1 labels the cuticular plates of hair cells, SC1 labels stereocilia and SP1 labels both stereocilia and pillar cells.…”
Section: Retinoic Acid Alters the Actin Cytoskeletonmentioning
confidence: 99%
“…Two exceptions are antibodies against the lateral cisternae of guinea pig outer hair cells (Holley and Richardson, 1994) and the circumferential actin ring (Nishida and Holley, 1996). Thus, we have not identified the antigens.…”
Section: Possible Antigens For Cp1 Sc1 and Sp1-sp3mentioning
confidence: 71%