Immunohistochemical Analysis of Proteoglycan Biosynthesis during Early Development of the Chicken Cornea.

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概要

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• Antibodies to core proteins of chicken corneal keratan sulfate proteoglycan and chondroitin sulfate proteoglycan were prepared and purified by use of an affinity column. Using these antibodies and monoclonal antibody 5-D-4 to keratan sulfate (commercial), the localization of proteoglycans in developing corneas (Days 5 to 17 of embryonic age and 2 days after hatching) was determined immunohistochemically. Keratan sulfate proteoglycan antigen was not detected in cornea on Day 5, but it was detected uniformly over the whole stroma on Day 6, ca. 12 h after invasion of the primary stroma by mesenchymal cells. The absence of the antigen in cornea of Day 5 was confirmed by Western blotting of the corneal extract. Immunohistochemistry with 5-D-4 antibody revealed that the keratan sulfate chain was undersulfated in corneas of Days 6 to 7, because the staining was much weaker than that in cornea of Day 8. In addition, keratan sulfate proteoglycan antigen was detected uniformly over the whole stroma on Days 7 to 17 and 2 days after hatching, but not in the epithelial layer on Day 13 and after: because the epithelial layer was clearly not observed on photomicrographs until Day 13, it is not known whether keratan sulfate proteoglycan was synthesized by the epithelium during Days 6 to 12. In contrast, chondroitin sulfate proteoglycan antigen was detected in cornea on Day 5 and also, like keratan sulfate proteoglycan, uniformly over the whole stroma on Day 6 through 2 days after hatching. Furthermore, the chondroitin sulfate proteoglycan was not detected in the epithelial layer on Day 13 and after. These results show that keratan sulfate proteoglycan is synthesized by the stromal cells which invade the primary stroma between Day 5.5 and 6, while chondroitin sulfate proteoglycan is synthesized by epithelial and/or endothelial cells before the invasion, and also by the stromal cells after the invasion.

• 社団法人 日本生化学会の論文

著者

• TAKAHASHI Ikuko

  Section of Radiochemistry, Faculty of Pharmacy, Meijo University

• Hamada Yuri

  Section Of Radiochemistry Faculty Of Pharmacy Meijo University

• Nakamura Youko

  Section Of Radiochemistry Faculty Of Pharmacy Meijo University

• Nakazawa Kiyoshi

  Section Of Radiochem. Fac. Of Pharm. Sci. Meijo Univ.

関連論文

• Comparative Analysis of Proteoglycans Synthesized by Chick Corneal Stromal Cells in Cell Culture and Organ Culture
• Immunohistochemical Analysis of Proteoglycan Biosynthesis during Early Development of the Chicken Cornea
• Proteoglycan Synthesis by Chicken Corneal Explants
• Histochemical detection of X-ray induced oxidative stress in mice by 2',7'-dichlorofluorescin
• Purification and Characterization of Chick Corneal β-_D-Glucuronyltransferase Involved in Chondroitin Sulfate Biosynthesis(Biochemistry/Molecular Biology)
• Modification of Proteoglycan Synthesis by Corneal Stromal Cells on Co-Culture with Either Epithelial or Endothelial Cells
• Keratan Sulfate Synthesis by Corneal Stromal Cells within Three-Dimensional Collagen Gel Cultures
• Proteoglycan Synthesis by Corneal Explants from Developing Embryonic Chicken
• Immunohistochemical Analysis of Proteoglycan Biosynthesis during Early Development of the Chicken Cornea.

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