Collagen metabolism during healing of lacerated rabbit corneas.
We previously demonstrated that there are two waves of increased collagen synthesis following corneal laceration in rabbits. In the present study, we have examined whether increases in collagen synthesis and degradation result from increased amounts of mRNAs for collagen and collagenase, respectively. Rabbits were anesthetized by combined administration of ketamine (intramuscular) and pentobarbital (intravenous). A penetrating 8-mm incision was made at the center of each cornea. The lacerated corneas were allowed to heal for 0-49 days. The rabbits were then killed and the corneas excised. The total RNA was extracted from the tissue and subjected to slot-blot hybridization using 32P-labeled alpha 1(I) cDNA. The results indicate that there is a two-phase increase in the amount of alpha 1(I) mRNA in injured corneas and that the collagenase mRNA is elevated at most times throughout the healing period. However, the increase is collagenase mRNA may not fully account for the accelerated collagen degradation during corneal wound-healing. Thus, we propose that cells in the wound area may be directly involved in collagen degradation by phagocytosis. To examine our hypothesis, we cultured injured rabbit corneas in the presence or absence of leupeptin, a proteinase inhibitor. The tissues were then examined by electron microscopy. In the presence of leupeptin, lysosomes within fibroblasts or fibroblast-like cells in the wound area of the lacerated corneas healed for 2 and 3 weeks, contain collagen fibrils. In the absence of leupeptin no identifiable collagen was seen in the lysosomes.(ABSTRACT TRUNCATED AT 250 WORDS)
Sakai
J
J
Department of Ophthalmology, University of cincinnati College of Medicine, OH 45267-0527.
Hung
J
J
Zhu
G
G
Katakami
C
C
Boyce
S
S
Kao
W W
WW
D000818:Animals; D003094:Collagen / Q000096:biosynthesis*; D003315:Cornea / Q000648:ultrastructure; D065306:Corneal Injuries; D005260:Female; D007976:Leupeptins / Q000494:pharmacology; D008247:Lysosomes / Q000187:drug effects / Q000648:ultrastructure; D008297:Male; D003012:Microbial Collagenase / Q000096:biosynthesis; D008854:Microscopy, Electron; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D013997:Time Factors; D014945:Wound Healing / Q000502:physiology*
D007976:Leupeptins; D012333:RNA, Messenger; D003094:Collagen; D003012:Microbial Collagenase