title: Gelatinase A and TIMP-2 expression in the fibrous sclera of myopic and recovering chick eyes.
abstract: OBJECTIVE
  Myopia, or nearsightedness, is characterized by excessive lengthening of the ocular globe and is associated with extracellular matrix remodeling in the posterior sclera. The activity of gelatinase A, a member of the matrix metalloproteinase family, has been shown to increase in the posterior sclera during the development of induced myopia in several species. In the present study, the distribution and relative expression of gelatinase A and its associated inhibitor, tissue inhibitor of metalloproteinases (TIMP)-2, were measured within the fibrous scleras of experimentally myopic (form-deprived) eyes, control eyes, and eyes recovering from form deprivation to better understand the mechanisms that regulate scleral remodeling and the rate of ocular elongation.
  
  
  METHODS
  Total RNA was extracted from the posterior scleras of form-deprived chick eyes, eyes recovering from deprivation myopia, and paired contralateral control eyes, and subjected to northern blot analysis analyses using cDNA probes to chicken gelatinase A and TIMP-2. The distribution of gelatinase A and TIMP-2 mRNAs was evaluated by in situ hybridization on frozen sections of chick scleras using 33P-labeled RNA probes. Gelatinase A activity within the fibrous scleras of form-deprived eyes and paired contralateral recovering eyes was evaluated by gelatin zymography.
  
  
  RESULTS
  Northern blot analysis indicated that the relative expression of gelatinase A was increased by 128% in deprived eyes (P = 0.009), whereas after 1 day of recovery, levels were decreased by 80% in scleras from recovering eyes (P = 0.005). In contrast, TIMP-2 expression was significantly decreased (-53%, P = 0.027) in the posterior scleras of form-deprived eyes. No significant differences were detected in levels of TIMP-2 expression between recovering eyes and paired control eyes. In situ hybridization indicated that most of the gelatinase A transcripts were present in the fibrous layer of the posterior scleras from form-deprived and recovering eyes.
  
  
  CONCLUSIONS
  Changes in the steady state levels of gelatinase A and TIMP-2 mRNA lead to changes in gelatinase activity within the fibrous sclera and mediate, at least in part, the process of visually regulated ocular growth and scleral remodeling.
authors.affiliation: Department of Anatomy and Cell Biology, University of North Dakota School of Medicine and Health Sciences, Grand Forks 58202, USA. jarada@medicine.nodak.edu
authors.forename: J A
authors.initials: JA
authors.lastname: Rada
authors.forename: C A
authors.initials: CA
authors.lastname: Perry
authors.forename: M L
authors.initials: ML
authors.lastname: Slover
authors.forename: V R
authors.initials: VR
authors.lastname: Achen
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D002645:Chickens; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression; D017403:In Situ Hybridization; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000235:genetics*; D009216:Myopia / Q000209:etiology / Q000378:metabolism* / Q000503:physiopathology; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012590:Sclera / Q000378:metabolism* / Q000503:physiopathology; D012683:Sensory Deprivation; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D019716:Tissue Inhibitor of Metalloproteinase-2; D012313:RNA; D020778:Matrix Metalloproteinase 2