Cochlin expression in anterior segment organ culture models after TGFbeta2 treatment.
OBJECTIVE
To determine the effect of transforming growth factor (TGF)-beta2 treatment on intraocular pressure (IOP), outflow facility, and cochlin expression in vitro in monkey and pig organ-cultured anterior segments (MOCAS and POCAS).


METHODS
MOCAS (rhesus and cynomolgus) or POCAS were infused with media containing 10 ng/mL TGFbeta2 to one segment of each pair and 0.1% BSA (vehicle) to the contralateral segment for up to 14 days at a constant rate. Cochlin expression was determined by immunohistochemical study, ELISA, and Western blot analysis using chicken polyclonal antibodies against different regions of cochlin.


RESULTS
TGFbeta2 infusion produced elevated IOP in MOCAS (usually after 5 days), that was approximately 45% greater than baseline and compared to control segments. Outflow facility (OF) was decreased by approximately 40% compared with pretreatment baseline (n=5). In POCAS (n=7), IOP was increased (approximately 3 days) by approximately 75% compared with baseline and contralateral changes. The IOP elevation subsided thereafter. Cochlin levels increased with duration of TGFbeta2 treatment in the media and in the region of the trabecular meshwork in both species.


CONCLUSIONS
TGFbeta2-induced IOP elevation was associated with an increase in cochlin secretion into the media and expression in the tissue of MOCAS and POCAS. Whether cochlin overexpression contributes to elevated IOP or is a consequence of other changes relevant to IOP elevation remains to be determined.
Bascom Palmer Eye Institute, University of Miami, Miami, Florida 33136, USA. sbhattacharya@med.miami.edu
Sanjoy K
SK
Bhattacharya
B'Ann T
BT
Gabelt
Jose
J
Ruiz
Renata
R
Picciani
Paul L
PL
Kaufman
D000818:Animals; D000869:Anterior Eye Segment / Q000187:drug effects* / Q000378:metabolism; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D016326:Extracellular Matrix Proteins; D019084:Fluorescent Antibody Technique, Indirect; D007429:Intraocular Pressure / Q000187:drug effects; D008252:Macaca fascicularis; D008253:Macaca mulatta; D009924:Organ Culture Techniques; D011506:Proteins / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D013552:Swine; D053781:Transforming Growth Factor beta2 / Q000494:pharmacology*
C110668:COCH protein, human; D016326:Extracellular Matrix Proteins; D011506:Proteins; D011994:Recombinant Proteins; C509041:TGFB2 protein, human; D053781:Transforming Growth Factor beta2