Type XIII collagen is widely expressed in the adult and developing human eye and accentuated in the ciliary muscle, the optic nerve and the neural retina.

The distribution of mRNAs coding for type XIII collagen, a novel nonfibril-forming collagen, was studied by Northern and in situ hybridizations of adult and fetal human eyes and the corresponding protein was localized by indirect immunofluorescence in frozen sections of 12 and 17 week human fetal eyes using a polyclonal antipeptide antibody to type XIII collagen. Type XIII collagen was found to be widely expressed in ocular tissues when studied at both the mRNA and protein levels in fetal and adult human tissues. No major differences were observed in the expression patterns between fetal and adult tissues. Surprisingly, the strongest signals seen in in situ hybridizations and immunofluorescence stainings occurred in the optic nerve bundles and in the ganglion cell layer of the retina. Other notable locations containing type XIII collagen included the developing ciliary smooth muscle, the posterior two-thirds of the corneal stroma and the striated extraocular muscles. Low level signals were also detected in the blood vessel walls and mesenchymal cells of the other ocular tissues. All immunosignals detected were adherent to cells, and the extracellular matrices appeared to be devoid of type XIII collagen. Our results are in concert with the presumed plasma membrane location of type XIII collagen, and it is hypothesized that this molecule could be involved in cell-matrix and perhaps cell-cell interactions. The wide expression of type XIII collagen in the eye, and especially in the neural structures, warrants future studies on type XIII collagen in other nerve structures and in pathological conditions affecting the eye. Due to its wide expression, type XIII collagen is likely to be an important factor for the normal development and functioning of the eye.

Type XIII collagen forms homotrimers with three triple helical collagenous domains and its association into disulfide-bonded trimers is enhanced by prolyl 4-hydroxylase.

Type XIII collagen is a type II transmembrane protein predicted to consist of a short cytosolic domain, a single transmembrane domain, and three collagenous domains flanked by noncollagenous sequences. Previous studies on mRNAs indicate that the structures of the collagenous domain closest to the cell membrane, COL1, the adjacent noncollagenous domain, NC2, and the C-terminal domains COL3 and NC4 are subject to alternative splicing. In order to extend studies of type XIII collagen from cDNAs to the protein level we have produced it in insect cells by means of baculoviruses. Type XIII collagen alpha chains were found to associate into disulfide-bonded trimers, and hydroxylation of proline residues dramatically enhanced this association. This protein contains altogether eight cysteine residues, and interchain disulfide bonds could be located in the NC1 domain and possibly at the junction of COL1 and NC2, while the two cysteine residues in NC4 are likely to form intrachain bonds. Pepsin and trypsin/chymotrypsin digestions indicated that the type XIII collagen alpha chains form homotrimers whose three collagenous domains are in triple helical conformation. The thermal stabilities (T(m)) of the COL1, COL2, and COL3 domains were 38, 49 and 40 degrees C, respectively. The T(m) of the central collagenous domain is unusually high, which in the light of this domain being invariant in terms of alternative splicing suggests that the central portion of the molecule may have an important role in the stability of the molecule. All in all, most of the type XIII collagen ectodomain appears to be present in triple helical conformation, which is in clear contrast to the short or highly interrupted triple helical domains of the other known collagenous transmembrane proteins.

Location of type XV collagen in human tissues and its accumulation in the interstitial matrix of the fibrotic kidney.

An antipeptide antibody was produced against the carboxyl-terminal noncollagenous domain of human type XV collagen and used to localize this recently described collagen in a number of human tissues. The most conspicuous findings were powerful staining of most of the capillaries and staining of the basement membrane (BM) zones of muscle cells. Not all of the BM zones were positive, however, as shown by the lack of staining in the developing fetal alveoli and some of the tubules in developing kidney. Nor was type XV collagen staining restricted to the BM zones, as some could be observed in the fibrillar collagen matrix of the papillary dermis and placental villi, for example. Interestingly, differences in the expression of type XV collagen could be observed during kidney development, and staining of fetal lung tissue suggested that changes in its expression may also occur during the formation of vascular structures. Another intriguing finding was pronounced renal interstitial type XV collagen staining in patients with kidney fibrosis due to different pathological processes. This suggests that the accumulation of type XV collagen may accompany fibrotic processes. Full-length human type XV collagen chains with an apparent molecular mass of approximately 200 kd were produced in insect cells using a baculovirus expression system. The fact that these had a markedly higher molecular mass than the 100- to 110-kd type XV collagen chains found in homogenates of heart and kidney tissue suggests either proteolytic processing during the synthesis of type XV collagen or an inability to solubilize complete molecules from tissues.