title: Effects of basic FGF and TGF beta 1 on F-actin and ZO-1 organization during cat endothelial wound healing.
abstract: Previous studies suggest the existence of two separate and distinct mechanisms of endothelial wound healing (i.e., cell migration and cell spreading), which may be controlled by unique, injury-dependent, wound-related factors. The purpose of our study was to evaluate potential biologic mediators regulating healing of the growth arrested cat endothelium by using an ex vivo, organ-culture model. Three buttons were punched from each cornea of 11 cats with a 6-mm trephine. A 1- to 2-mm diameter endothelial scrape injury (SI) was made, and buttons were cultured in (a) serum-free media (SFM), (b) serum plus media (20% fetal calf serum), (c) SFM plus basic fibroblast growth factor (bFGF), (d) SFM plus bFGF and heparin, (e) SFM plus transforming growth factor-beta 1 (TGF beta 1), or (f) SFM plus TGF beta 1 and anti-TGF beta 1. At various times from 8-48 h after injury, buttons were stained with phalloidin and anti-ZO-1, and imaged by using laser scanning confocal microscopy. Evaluation of SI in cat corneal buttons under serum-free conditions showed maintenance of normal endothelial differentiation, indicating that the organ-culture SI model mimics in vivo SI. Addition of TGF beta 1 produced a dramatic reorganization of apical F-actin and development of stress fibers, as well as the loss of normal cell border-associated ZO-1 distribution. The effects of TGF beta 1 were blocked by the neutralizing antibodies to TGF beta 1. Addition of serum or bFGF produced much less pronounced changes in F-actin and ZO-1 distribution. These results suggest that TGF beta 1 may play a critical role in modulating the wound-healing response of the corneal endothelium.
authors.affiliation: Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas 75235-9057, USA.
authors.forename: W M
authors.initials: WM
authors.lastname: Petroll
authors.forename: J V
authors.initials: JV
authors.lastname: Jester
authors.forename: P A
authors.initials: PA
authors.lastname: Barry-Lane
authors.forename: H D
authors.initials: HD
authors.lastname: Cavanagh
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D001769:Blood; D002415:Cats; D016895:Culture Media, Serum-Free; D004728:Endothelium, Corneal / Q000293:injuries / Q000378:metabolism* / Q000473:pathology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D019084:Fluorescent Antibody Technique, Indirect; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D009924:Organ Culture Techniques; D010750:Phosphoproteins / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D014945:Wound Healing / Q000502:physiology*; D062826:Zonula Occludens-1 Protein
chemical_list: D000199:Actins; D016895:Culture Media, Serum-Free; D008565:Membrane Proteins; D010750:Phosphoproteins; D016212:Transforming Growth Factor beta; D062826:Zonula Occludens-1 Protein; D016222:Fibroblast Growth Factor 2