PURPOSE: To determine whether trabecular tissue in vivo and cultured trabecular cells have the messenger RNA transcript for transforming growth factor-beta 1 (TGF-beta 1), and to examine whether these cells synthesize and secrete TGF-beta 1 in vitro. METHODS: Total RNA was isolated from the trabecular meshwork, iris, and ciliary body freshly excised from porcine eyes as well as from cultured trabecular cells, and the reverse transcriptase-polymerase chain reaction and Southern hybridization were used for detection of TGF-beta 1 messenger RNA. The amount of TGF-beta 1 secreted by trabecular cells in culture was determined by radioimmunoassay. RESULTS: Excised whole trabecular tissue, iris, and ciliary body, as well as cultured trabecular cells expressed messenger RNA transcripts for TGF-beta 1. On the ethidium bromide-stained agarose gel, two PCR-amplified products (161 and 400 base pairs) were found in the total RNA isolated from cultured trabecular cells. The oligonucleotide probe specific for TGF-beta 1 detected only one band with the expected length of 161 base pairs. The secretion of TGF-beta 1 into conditioned medium was at the level of 16.7-20 pg/ml per 2 million trabecular cells during a 24-hr period. CONCLUSIONS: These investigations show that the trabecular meshwork, iris, and ciliary body in vivo express the messenger RNA transcript for TGF-beta 1, and that trabecular cells in vitro synthesize and secrete this cytokine. The TGF-beta 1 present in normal aqueous humor may be derived locally, at least in part, from the cells of the trabecular meshwork, iris, and ciliary body. Abnormal synthesis, secretion, activation, and clearance of TGF-beta 1 may contribute to the pathogenesis of many ocular disorders, including primary open-angle glaucoma.
PURPOSE: To determine whether trabecular tissue in vivo and cultured trabecular cells have the messenger RNA transcript for transforming growth factor-beta 1 (TGF-beta 1), and to examine whether these cells synthesize and secrete TGF-beta 1 in vitro. METHODS: Total RNA was isolated from the trabecular meshwork, iris, and ciliary body freshly excised from porcine eyes as well as from cultured trabecular cells, and the reverse transcriptase-polymerase chain reaction and Southern hybridization were used for detection of TGF-beta 1 messenger RNA. The amount of TGF-beta 1 secreted by trabecular cells in culture was determined by radioimmunoassay. RESULTS: Excised whole trabecular tissue, iris, and ciliary body, as well as cultured trabecular cells expressed messenger RNA transcripts for TGF-beta 1. On the ethidium bromide-stained agarose gel, two PCR-amplified products (161 and 400 base pairs) were found in the total RNA isolated from cultured trabecular cells. The oligonucleotide probe specific for TGF-beta 1 detected only one band with the expected length of 161 base pairs. The secretion of TGF-beta 1 into conditioned medium was at the level of 16.7-20 pg/ml per 2 million trabecular cells during a 24-hr period. CONCLUSIONS: These investigations show that the trabecular meshwork, iris, and ciliary body in vivo express the messenger RNA transcript for TGF-beta 1, and that trabecular cells in vitro synthesize and secrete this cytokine. The TGF-beta 1 present in normal aqueous humor may be derived locally, at least in part, from the cells of the trabecular meshwork, iris, and ciliary body. Abnormal synthesis, secretion, activation, and clearance of TGF-beta 1 may contribute to the pathogenesis of many ocular disorders, including primary open-angle glaucoma.
Authors: Nikoleta Tellios; Jillian C Belrose; Alexander C Tokarewicz; Cindy Hutnik; Hong Liu; Andrew Leask; Michael Motolko; Miho Iijima; Sunil K Parapuram Journal: Sci Rep Date: 2017-04-11 Impact factor: 4.379