Upregulation of bone morphogenetic protein-1/mammalian tolloid and procollagen C-proteinase enhancer-1 in corneal scarring.

Investigative ophthalmology & visual science (2014-09-25)
Francois Malecaze, Dawiyat Massoudi, Pierre Fournié, Cyrielle Tricoire, Myriam Cassagne, Marilyne Malbouyres, David J S Hulmes, Catherine Moali, Stephane D Galiacy
RESUMEN

To characterize the expression of the bone morphogenetic protein-1 (BMP-1)/tolloid-like proteinases (collectively called BTPs), which include BMP-1, mammalian tolloid (mTLD), and mammalian tolloid-like 1 (mTLL-1) and 2 (mTLL-2), as well as the associated proteins procollagen C-proteinase enhancers (PCPE-1 and -2), in corneal scarring. Using a mouse full-thickness corneal excision model, wound healing was followed for up to 28 days by transmission electron microscopy, immunohistology (BMP-1/mTLD and PCPE-1), and quantitative PCR (Q-PCR: collagen III, BMP-1/mTLD, mTLL-1, mTLL-2, PCPE-1, PCPE-2). Bone morphogenetic protein-1/mTLD and PCPE-1 were also immunolocalized in cases of human corneal scarring following injuries. In the mouse model, throughout the follow-up period, there was a large increase in collagen III mRNA expression in the stroma. By transmission electron microscopy, there was marked cellular infiltration into the wound as well as disorganization of collagen fibrils, but no significant difference in fibril diameter. In control corneas, by Q-PCR, BMP-1/mTLD showed the highest expression, compared to low levels of mTLL-1 and undetectable levels of mTLL-2, in both epithelium and stroma. Following wounding, both BMP-1/mTLD and PCPE-1 mRNA and protein increased, while PCPE-2 mRNA decreased. Finally, by immunofluorescence, BMP-1/mTLD and PCPE-1 were strongly expressed in the scar region in both mouse and human corneas. Bone morphogenetic protein-1/mTLD and PCPE-1 are upregulated in corneal scars. Both proteins may therefore contribute to the process of corneal scarring.

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Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Rabbit serum
Supelco
Atropine Sulfate, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Xylazine hydrochloride, ≥99.0% (HPLC)
USP
Atropine sulfate, United States Pharmacopeia (USP) Reference Standard
Supelco
Xylazine hydrochloride, VETRANAL®, analytical standard
Atropine sulfate, European Pharmacopoeia (EP) Reference Standard
Xylazine hydrochloride, European Pharmacopoeia (EP) Reference Standard

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