December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Immunohistochemical Characterization of Developing Eyes of Type XVIII Collagen Knockout Mice
Author Affiliations & Notes
  • JD Javier
    Ophthalmology Massachusetts Eye and Ear Infirmary and Schepens Eye Research Institute Boston MA
  • JB Lee
    Ophthalmology Massachusetts Eye and Ear Infirmary and Schepens Eye Research Institute Boston MA
  • JH Chang
    Ophthalmology Massachusetts Eye and Ear Infirmary and Schepens Eye Research Institute Boston MA
  • T Kato
    Ophthalmology Massachusetts Eye and Ear Infirmary and Schepens Eye Research Institute Boston MA
  • N Fukai
    Cell Biology Harvard Medical School Boston MA
  • B Olsen
    Cell Biology Harvard Medical School Boston MA
  • DT Azar
    Ophthalmology Massachusetts Eye and Ear Infirmary and Schepens Eye Research Institute Boston MA
  • NF Azar
    Ophthalmology Massachusetts Eye and Ear Infirmary and Schepens Eye Research Institute Boston MA
  • Footnotes
    Commercial Relationships   J.D. Javier, None; J.B. Lee, None; J.H. Chang, None; T. Kato, None; N. Fukai, None; B. Olsen, None; D.T. Azar, None; N.F. Azar, None. Grant Identification: NIH EY10101
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 3221. doi:
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    • Get Citation

      JD Javier, JB Lee, JH Chang, T Kato, N Fukai, B Olsen, DT Azar, NF Azar; Immunohistochemical Characterization of Developing Eyes of Type XVIII Collagen Knockout Mice . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3221.

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Abstract

Abstract: : Purpose: To immunolocalize metalloproteinases, collagens and antiangiogenic factors in Type XVIII collagen knockout mice during development and wound healing. Methods: Littermate offspring from heterozygous and homozygous collagen XVIII knockout C57BL/6 mice were included in this study. Mice were genotyped using PCR, and the eyes mounted in OCT and sectioned for immunohistochemistry. Specimens from heterozygous and homozygous collagen XVIII knockout mice were used to determine the immunolocalization of collagen IV, collagen XV A, collagen XV B, NC-11 and hinge domain (AbN) of collagen XVIII, MMP-7, PEDF, angiostatin (K1) and plasminogen (B chain) using well- characterized polyclonal antibodies. Specimens were examined by immunofluorescence and confocal microscopy. Results: No phenotypic anatomic abnormalities were observed in the eyes of the knockout mice. NC-11 was immunolocalized to the posterior lid margin and corneal epithelium and epithelial basement membrane in heterozygous animals and was absent in knockout animals. Type IV collagen was primarily localized in the palpebral conjunctiva, anterior and posterior layers of the cornea and in the lens capsule. AbN staining was noted in the cornea and lids in both heterozygous and homozygous knockout animals. There were no differences in the immunolocalization patterns of the other enzymes, collagens and antiangiogenic factors between the wild type and knockout mice. Conclusion: The immunohistochemical alterations in the homozygous collagen type XVIII knockout mice do not seem to be associated with phenotypic abnormalities.

Keywords: 370 cornea: basic science • 483 neovascularization • 316 animal model 
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