A protein of 175,000 daltons associated with striated rootlets in ciliated epithelia, as revealed by a monoclonal antibody.
Basal bodies from laying quail oviduct were semipurified and used as immunogen to produce monoclonal antibodies. On 38 clones obtained and among those staining the apical pole of the ciliated cell, CC-310 was chosen because it labeled the apical region with a punctuated aspect, suggesting a staining of basal bodies or of basal body-associated structures; the basal pole was also labeled. The ultrastructural localization performed by the immunogold technique showed that the labeling was mainly associated with the striated rootlets. The basal feet, the side of the basal bodies, and the basal poles of the demembranated cells were also decorated. The identification of the antigen performed by immunoblots of deciliated cortices revealed two proteins of 175,000 and 40,000, whereas immunoblots of basal bodies showed only the 175,000-mw protein. The possibility of these two proteins sharing the same epitope, located at both poles of the cell, is discussed. Immunofluorescence ascertained that CC-310 decorated the striated rootlets in ciliated epithelia from other species: mussel, frog, and human tissue. Finally, when tested on cultured cell lines, CC-310 labeled the centrosome and its associated rootlets on PtK2 during interphase. During mitosis the poles of the mitotic spindle were stained without any apparent rootlet-like structure.
Klotz
C
C
Bordes
N
N
Laine
M C
MC
Sandoz
D
D
Bornens
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis / Q000276:immunology; D049872:Bivalvia; D002460:Cell Line; D002923:Cilia / Q000032:analysis / Q000648:ultrastructure; D003370:Coturnix; D006224:Cricetinae; D003598:Cytoskeletal Proteins / Q000032:analysis* / Q000276:immunology; D004848:Epithelium / Q000032:analysis / Q000648:ultrastructure; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007614:Macropodidae; D051379:Mice; D008815:Mice, Inbred Strains; D008854:Microscopy, Electron; D008970:Molecular Weight; D010057:Oviducts / Q000166:cytology*; D011893:Rana esculenta; D013045:Species Specificity; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D000941:Antigens; D003598:Cytoskeletal Proteins