A characteristics protein highly expressed in guinea pig inner ear, defined by monoclonal antibody WH-1.
A new monoclonal antibody (termed WH-1; isotype IgG2b) was established using a homogenate of dissected guinea pig cochleas (N = 60) as immunogen. Western blotting and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) identified the WH-1 antigen as a protein or glycoprotein with M(r) approximately 40 kDa. Immunoperoxidase treatment of histologic cryosections of guinea pig cochlea, followed by light microscopic examination, revealed strong positive staining at three sites: (i) parts of Hensen's stripe, marginal band, covering net, and Kimura's membrane (within the tectorial membrane [TM]); (ii) Deiters' cells, pillar cells, Hensen's cells, and stereocilia of outer hair cells (within the organ of Corti); (iii) interdental cells, inner and outer sulcus cells, Reissner's membrane, and surface membrane of stria vascularis epithelium. Similar staining patterns were observed for cryosections of rat and mouse cochleas. Only a trace quantity of cross-reacting protein was detectable in brainstem. The protein was not detectable in tongue extract by Western blotting. However, sections of brainstem and tongue did show positive immunohistological staining with WH-1. Localization of WH-1 antigen was further examined by electron microscopy. WH-1 positivity on outer hair cell stereocilia, certain sites on the TM, interdental cell surface, Reissner's membrane epithelia, and inner and outer sulcus cells was confirmed. WH-1 antigen was not detected on inner hair cell stereocilia by light or electron microscopy. The localization of WH-1 antigen on outer hair cell stereocilia and TM suggests that it may play some role in adhesion between these structures.
Wataya
H
H
Biomembrane Institute, Seattle, WA 98119.
Handa
K
K
Hozawa
K
K
Takasaka
T
T
Cunningham
D
D
Rubel
E W
EW
Hakomori
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D000941:Antigens / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D003051:Cochlea / Q000378:metabolism* / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D006168:Guinea Pigs; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D008970:Molecular Weight; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry / Q000276:immunology; D014611:Vaccination
D000911:Antibodies, Monoclonal; D000941:Antigens; D006023:Glycoproteins; D011506:Proteins