Expression of VSX1 in human corneal keratocytes during differentiation into myofibroblasts in response to wound healing.
OBJECTIVE
To characterize the expression of the visual system homeobox gene (VSX1) in human corneal keratocytes both in vitro and in vivo.


METHODS
The expression of VSX1 was evaluated through semiquantitative RT-PCR, immunofluorescence and in situ hybridization both in corneas (either freshly obtained or wounded) and in collagenase/hyaluronidase-isolated keratocytes grown in the absence or presence of serum to promote keratocyte-to-myofibroblast differentiation.


RESULTS
Quiescent or resting keratocytes normally residing in the corneal stroma or cultured in vitro in the absence of serum did not express VSX1. In wounded corneas or when cultured in the presence of serum to mimic wound-healing responses, keratocytes underwent fibroblastic transformation (with appearance of alpha-SMA and disappearance of CD-34 and keratocan signals) and started expressing VSX1.


CONCLUSIONS
The results show that VSX1 is expressed in vitro and in vivo during human corneal wound healing, a process in which differentiation of corneal keratocytes into myofibroblasts occurs. These data may help to elucidate the role of VSX1 in cornea physiology suggesting a potential involvement in cornea-related diseases such as keratoconus.
Barbaro
Vanessa
V
Epithelial Stem Cell Research Centre, Veneto Eye Bank Foundation, Venice, Italy; the.
Di Iorio
Enzo
E
Ferrari
Stefano
S
Bisceglia
Luigi
L
Ruzza
Alessandro
A
De Luca
Michele
M
Pellegrini
Graziella
G
D000199:Actins / Q000378:metabolism; D018952:Antigens, CD34 / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D003319:Corneal Stroma / Q000166:cytology*; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D017403:In Situ Hybridization; D008856:Microscopy, Fluorescence; D011509:Proteoglycans / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014945:Wound Healing / Q000502:physiology*
D000199:Actins; D018952:Antigens, CD34; D005136:Eye Proteins; D018398:Homeodomain Proteins; C490948:KERA protein, human; D011509:Proteoglycans; D012333:RNA, Messenger; C491583:VSX1 protein, human