Changes in distribution of the long form of type XII collagen during chicken corneal de.pdf

? The Histochemical Society, Inc. 0022-1554/02/$3.30 851 ARTICLE Volume 50(6): 851–862, 2002 The Journal of Histochemistry Cytochemistry Changes in Distribution of the Long Form of Type XII Collagen during Chicken Corneal Development Yoshihiro Akimoto, Naomi Yamakawa, Kiyoshi Furukawa, Koji Kimata, Hayato Kawakami, and Hiroshi Hirano Department of Anatomy, Kyorin University School of Medicine, Tokyo, Japan (YA,HK,HH); Department of Biosignal Research, Tokyo Metropolitan Institute of Gerontology, Tokyo, Japan (NY,KF); Institute of Medical Science, Aichi Medical College, Aichi, Japan (KK); and Nittai Jusei Medical College for Judo Therapeutics, Tokyo, Japan (HH) SUMMARY The expression and distribution of the long form of Type XII collagen were in- vestigated histochemically during chicken corneal development using a monoclonal anti- body (P3D11) raised against the N-terminal domain of chicken Type XII collagen. Specificity of the antibody was confirmed by immunoprecipitation before and after bacterial collage- nase digestion. Immunofluorescent microscopic studies showed that during chicken cornea formation, the long form of Type XII collagen is initially detected on Day 3 embryo (stage 19) in the sub-epithelial matrix of the corneal periphery and in the matrix around the optic cup. On Day 5 embryo (stage 27) the long form was expressed in the primary stroma. There- after, as the secondary stroma was formed, the long form localized in the sub-epithelial and sub-endothelial matrices and in the anterior region of the limbus (corneoscleral junc- tion) before the formation of Descemet’s and Bowman’s membranes. After hatching, the immunoreactivity decreased predominantly in the sub-epithelial and sub-endothelial ma- trices but remained at the anterior region of the limbus. Immunoelectron microscopic ex- amination demonstrated that the long form localizes in the Descemet’s and Bowman’s membranes and along the collagen fibrils in the stroma with a p