Proteoglycan turnover in the sclera of normal and experimentally myopic chick eyes.
OBJECTIVE
The turnover of chick scleral proteoglycans from control and form-vision deprived (myopic) eyes was compared in vivo and in explant cultures to determine whether proteoglycan degradation is altered during the development of myopia and to characterize the mechanism of proteoglycan turnover in the sclera.


METHODS
Seven-day-old chicks were radiolabeled via an intraperitoneal injection of 35SO4, and monocular form deprivation was induced 48 hours later. After 1, 2, and 3 weeks of form deprivation, birds were killed, and the amount of 35SO4-proteoglycans remaining in different scleral regions was measured in control and deprived eyes. Posterior sclera were also radiolabeled in organ culture containing 35SO4, and radiolabeled scleral proteoglycans were chased into unlabeled medium for 0 to 11 days. 35SO4-labeled proteoglycans within the scleral matrix and those released into the medium were characterized by Sepharose CL-2B chromatography and western blot analysis.


RESULTS
The biological half-life of scleral proteoglycans was significantly shorter within the posterior pole of form-deprived eyes (t1/2 = 7.212 days) compared with the same region of control eyes (t1/2 = 9.619 days; P < 0.001), whereas no differences in turnover rates were seen in the anterior sclera or equatorial sclera. When posterior scleral punches were placed in organ culture, 35SO4-labeled proteoglycan turnover rates were similar for control and form-deprived eyes. Chromatographic and western blot analyses indicated that approximately 80% of the total 35SO4 within the posterior sclera is incorporated into the aggrecan. Western blot analyses of aggrecan core protein released into the medium by control and form-deprived scleral punches indicated that the core protein was degraded into a series of smaller fragments of Mr = 102 to 220 kDa. A specific antiserum (anti-FVDIPEN) detected the presence of a 50-kDa C-terminal aggrecan fragment released into the medium, which was generated by the action of the matrix metalloproteinase gelatinase A and/or stromelysin.


CONCLUSIONS
The turnover rate of 35SO4-labeled scleral proteoglycans is vision dependent and is accelerated in the posterior sclera of chick eyes during the development of experimental myopia. The loss of proteoglycans from the scleral matrix involves proteolytic cleavage at various sites along the aggrecan core protein through the action, at least in part, of gelatinase A and/or stromelysin.
Department of Anatomy and Cell Biology, University of North Dakota School of Medicine and Health Sciences, Grand Forks 58202, USA.
J A
JA
Rada
V R
VR
Achen
K G
KG
Rada
D053674:Aggrecans; D000818:Animals; D015153:Blotting, Western; D002645:Chickens; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D002850:Chromatography, Gel; D016326:Extracellular Matrix Proteins; D018093:Gelatinases / Q000494:pharmacology; D006207:Half-Life; D037181:Lectins, C-Type; D008297:Male; D020778:Matrix Metalloproteinase 2; D019278:Matrix Metalloproteinase 3 / Q000494:pharmacology; D008666:Metalloendopeptidases / Q000494:pharmacology; D009216:Myopia / Q000209:etiology / Q000378:metabolism*; D009924:Organ Culture Techniques; D010446:Peptide Fragments / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism*; D012590:Sclera / Q000187:drug effects / Q000378:metabolism*; D012683:Sensory Deprivation
D053674:Aggrecans; D011508:Chondroitin Sulfate Proteoglycans; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D010446:Peptide Fragments; D011509:Proteoglycans; D018093:Gelatinases; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2