356 Fifth International Conference on the Molecular Biology and Pathology of Matrix

labeled for the type XV mRNA. There was a clear and relatively strong signal for the type XVIII collagen mRNA in fibroblastic cells around devel- oping bone and in developing muscle cells. A relatively weak signal was seen in hepatocytes of liver. The epithelial cells of collecting ducts in kidney were markedly labeled also for the type XVIII collagen mRNA. Although the cDNA- derived sequences show strong homology between types XV and XVIII, the Northern and in-situ analyses indicate that they are expressed partly in different tissues and in markedly different levels in some of the tissues where both are expressed.

gene to the syntenic region of chromosome 19. The deduced polypeptide has a C-terminal collage- nous domain that consists of 13 separate segments. Immunofluorescence analyses have indicated that this collagenous domain resides in the extracellular milieu, while the N-terminal non-collagenous do- main of 573 amino acids resides within the cyto- plasm of basal keratinocytes. Thus, type XVII collagen demonstrates an unusual type II trans- membrane topography. Type XVII collagen serves as an autoantigen in some patients with bullous pemphigoid, an acquired blistering disease, and was previously designated as the 180-kDa bullous pemphigoid antigen. Thus, type VII and XVII col- lagens are associated with BMZ pathology.

Cloning of Collagens of the Cutaneous Basement Membrane Zone: Types VII and X V I I

Jouni Uitto*, Angela M. Christiano*, Daniel S. Greenspant and Kehua Li*

*Departments of Dermatology and Biochemistry and Molecular Biology, Jefferson Medical College, Philadelphia, PA 19107; and tDepartment of Pathology and Laboratory Medicine, University of Wisconsin, Madison, WI 53708, USA

Collagens comprise a family of closely related, yet genetically distinct proteins, designated as types I- XIX. At least 11 of these collagens are found in the skin, and two of the collagens, types VII and XVII, are expressed in the basement membrane zone (BMZ) of stratified squamous epithelia. We have cloned the full-length human type VII collagen cDNA, as well as the entire gene (COL7A1) which we have mapped to human chromosome 3p21. The entire gene spans approximately 31 kb of the genomic DNA, and consists of 118 separate exons, the largest number reported in any gene. The de- duced amino acid sequence of the coding region indicates that type VII collagen consists of a central collagenous domain interrupted by 19 imperfec- tions and non-collagenous insertions. The large N- terminal NC-1 domain consists of submodules with homology to known adhesive proteins, and the smaller NC-2 domain has homology to the Kunitz protease inhibitor. Mutations in COL7A1 have been disclosed in the dystrophic forms of epider- molysis bullosa.

Type XVII collagen is a 180-kDa transmembrane protein localized to hemidesmosomes in the cuta- neous BMZ. We have cloned the full-length type XVII collagen cDNA, and assigned the human gene to chromosomal locus 10q24.3 and the mouse

2. New Information about Old Collagens

Localization of Basement Membrane Collagen Complex in Liver and Its Concentration in Sera after Chronic Administration of Carbon Tetrachloride

E. Adachi, T. Yoshida, T. Nagura, Y. Masuda and T. Hayashi

Medical School Department of Anatomy, Osaka University, Suita; Shiseido Research Center, Yokohama; and The University of Tokyo College of Arts and Sciences Department of Chemistry, Tokyo, Japan

We attempted to clarify the causal relationships between accumulation of collagen fibrils and of basement membrane collagen complex (BCC) rec- ognized by a monoclonal antibody JK-132 in fibrotic liver of crab-eating monkeys induced by administration of CC14. Liver tissues were obtained through biopsy under a laparoscope at intervals of 4 weeks. Simultaneously, monkey sera were also collected and the concentration of CEB measured by sandwich ELISA using two monoclonal anti- bodies (JK-199 and JK-132).

At 2 to 3 months after injection of CC14 (0.1-0.6 mg/kg body weight) twice a week, the concentra- tion of BCC increased 2 to 3 times as high as that under normal conditions. Histologically, fibrotic septa were observed in liver lobules at this stage. No basal lamina, however, was found under the endothelial cells.

At 5 to 10 months after CC14 administration, the concentration of BCC decreased to normal level. No significant sign of liver fibrosis was observed in biopsy samples. However, immunofluorescence