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Transforming growth factor‐β1 expression in cultured corneal fibroblasts in response to injury
Author(s) -
Song Qin Hui,
Singh Rishi P.,
Richardson Thomas P.,
Nugent Matthew A.,
TrinkausRandall Vickery
Publication year - 2000
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(20000501)77:2<186::aid-jcb3>3.0.co;2-m
Subject(s) - wound healing , transforming growth factor , messenger rna , northern blot , stromal cell , receptor , gene expression , in situ hybridization , microbiology and biotechnology , biology , chemistry , immunology , gene , cancer research , genetics
The mechanisms underlying TGF‐β regulation in response to injury are not fully understood. We have developed an in vitro wound model to evaluate the expression and localization of transforming growth factor‐β1 in rabbit corneal fibroblasts in response to injury. Experiments were conducted in the presence or absence of serum so that the effect of the injury could be distinguished from exogenous wound mediators. Cultures were wounded and evaluations conducted over a number of time points. Expression of TGF‐β1 RNA was determined using Northern blot analysis and in situ hybridization, while the TGF‐β receptors were identified by affinity cross‐linking. Injury increased the expression of TGF‐β1 mRNA in cells at the wound edge after 30 min; this response was amplified by the addition of serum. TGF‐β1 mRNA expression was observed in a number of cells distal from the wound. After wound closure, TGF‐β1 mRNA was negligible and resembled unwounded cultures. The half‐life of TGF‐β1 mRNA was two times greater in the wounded cultures, indicating that the injury itself maintained the expression, while cell migration was present. Analogous to these findings, we found that binding of TGF‐β to its receptors was maximal at the wound edge, decreasing with time and distance from the wound. These results indicate that injury increases the level of expression of TGF‐β1 mRNA and maintains a higher level of receptor binding during events in wound repair and that these might facilitate the migratory and synthetic response of stromal fibroblasts. J. Cell. Biochem. 77:186–199, 2000. © 2000 Wiley‐Liss, Inc.