title: Immunohistochemical and Western Blotting Analyses of Ganoine in the Ganoid Scales of Lepisosteus oculatus: an Actinopterygian Fish.
abstract: In order to compare its characteristics with those of jaw tooth collar enamel, normally developing and experimentally regenerating ganoine from ganoid scales of Lepisosteus oculatus (spotted gar), an actinopterygian fish species, was examined by Western blotting and immunohistochemistry. Amelogenin, a major enamel matrix protein (EMP), is widely found from sarcopterygian fish to mammals. Therefore, we used antimammalian amelogenin antibodies and antisera: an antibody against bovine amelogenin; antiserum against porcine amelogenin; and region-specific antibodies or antiserum against the C-terminus, middle region, or N-terminus of porcine amelogenin in this study. Positive immunoreactivity with the antibody against bovine amelogenin, antiserum against porcine amelogenin, and the middle and C-terminal region-specific antibodies was detected in both normally developing and regenerating ganoine matrix, as well as in granules found within inner ganoine epithelial cells. These immunohistochemical analyses indicated that the Lepisosteus ganoine matrix contains EMP-like proteins with epitopes similar to mammalian amelogenins. In Western blotting analyses of regenerating ganoid scales with the antibovine amelogenin antibody, two protein bands with molecular weights of approximately 78 and 65 kDa were detected, which were similar to those found in Lepisosteus tooth enamel. Our study suggests that in Lepisosteus, EMP-like proteins in the ganoine matrix corresponded to those in tooth enamel. However, it was revealed that the 78 and 65 kDa EMP-like proteins were different from 27 kDa bovine amelogenin.
authors.affiliation: Advanced Research Center, School of Life Dentistry at Niigata, The Nippon Dental University, Niigata, Japan.
authors.forename: Ichiro
authors.initials: I
authors.lastname: Sasagawa
authors.affiliation: Department of Biology, School of Life Dentistry at Niigata, The Nippon Dental University, Niigata, Japan.
authors.forename: Shunya
authors.initials: S
authors.lastname: Oka
authors.affiliation: Department of Microbiology, School of Life Dentistry at Niigata, The Nippon Dental University, Niigata, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Mikami
authors.affiliation: Department of Histology, School of Life Dentistry at Niigata, The Nippon Dental University, Niigata, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yokosuka
authors.affiliation: Department of Histology, School of Life Dentistry at Niigata, The Nippon Dental University, Niigata, Japan.
authors.forename: Mikio
authors.initials: M
authors.lastname: Ishiyama
authors.affiliation: Department of Biochemistry, School of Life Dentistry at Niigata, The Nippon Dental University, Niigata, Japan.|Department of Dental Hygiene, College at Niigata, The Nippon Dental University, Niigata, Japan.
authors.forename: Akane
authors.initials: A
authors.lastname: Imai
authors.affiliation: Division of Pediatric Dentistry, Department of Oral Health Sciences, Graduate School, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan.
authors.forename: Hitoyata
authors.initials: H
authors.lastname: Shimokawa
authors.affiliation: Department of Oral Biology, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Uchida
mesh_terms: D053523:Amelogenin / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D003743:Dental Enamel / Q000378:metabolism; D003746:Dental Enamel Proteins / Q000378:metabolism*; D005399:Fishes / Q000378:metabolism*; D007150:Immunohistochemistry; D012867:Skin / Q000254:growth & development / Q000378:metabolism*
chemical_list: D053523:Amelogenin; D003746:Dental Enamel Proteins