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Tissue-specific expression of type XII collagen during mouse embryonic development
S P Oh1, C M Griffith, E D Hay
1Department of Anatomy and Cellular Biology, Harvard Medical School, Boston, Massachusetts 02115.
Summary
Researchers developed polyclonal antibodies to study mouse type XII collagen, a protein found in dense connective tissues. This protein exists in two forms, with cultured cells primarily producing the larger 350 kDa version.
Area of Science:
- Biochemistry
- Molecular Biology
- Developmental Biology
Background:
- Type XII collagen is a fibril-associated collagen involved in connective tissue organization.
- Understanding the expression patterns and forms of type XII collagen is crucial for comprehending tissue development and integrity.
Purpose of the Study:
- To generate specific antibodies for detecting mouse type XII collagen.
- To investigate the expression sites and molecular forms of type XII collagen in mouse tissues and cultured cells.
Main Methods:
- Generation of polyclonal antibodies against a fusion peptide of mouse type XII collagen.
- Western blotting of mouse tissue extracts and cultured fibroblast medium.
- Immunohistochemical analysis of mouse embryos.
Main Results:
- Antibodies recognized 220 kDa and 350 kDa polypeptides in mouse tissue extracts.
- Type XII collagen expression was detected in dense connective tissues including tendons, ligaments, dermis, cornea, blood vessel walls, meninges, and developing membranous bones.
- Cultured skin fibroblasts predominantly produced the 350 kDa (long) form, while tissue extracts contained both 220 kDa (short) and 350 kDa forms.
Conclusions:
- Specific antibodies successfully detected type XII collagen in mouse tissues.
- Type XII collagen is widely distributed in dense connective tissues during embryonic development.
- Differential expression of type XII collagen forms occurs between native tissues and cultured fibroblasts.