Transforming growth factor-beta1 expression in cultured corneal fibroblasts in response to injury.
The mechanisms underlying TGF-beta 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-beta1 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-beta1 RNA was determined using Northern blot analysis and in situ hybridization, while the TGF-beta receptors were identified by affinity cross-linking. Injury increased the expression of TGF-beta1 mRNA in cells at the wound edge after 30 min; this response was amplified by the addition of serum. TGF-beta1 mRNA expression was observed in a number of cells distal from the wound. After wound closure, TGF-beta1 mRNA was negligible and resembled unwounded cultures. The half-life of TGF-beta1 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-beta 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-beta1 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.
Song
Q H
QH
Departments of Ophthalmology and Biochemistry, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
Singh
R P
RP
Richardson
T P
TP
Nugent
M A
MA
Trinkaus-Randall
V
V
D000818:Animals; D002478:Cells, Cultured; D003315:Cornea / Q000378:metabolism*; D065306:Corneal Injuries; D003470:Culture Media; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D066298:In Vitro Techniques; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D016212:Transforming Growth Factor beta / Q000235:genetics* / Q000378:metabolism*
D003470:Culture Media; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta