Injury induces endothelial to mesenchymal transition in the mouse corneal endothelium in vivo via FGF2

• Published in: Molecular vision 2019, Vol.25 (1), p.22-34
• Main Authors: Lee, JeongGoo, Jung, Eric, Heur, Martin
• Format: Article
• Language: English
• Subjects: Actin; Animals; Aqueous Humor - chemistry; Aqueous Humor - metabolism; Cdh1 Proteins - genetics; Cdh1 Proteins - metabolism; Collagen (type I); Collagen Type I - genetics; Collagen Type I - metabolism; Cornea; Cornea - metabolism; Cornea - pathology; corneal endothelium; Corneal Injuries - genetics; Corneal Injuries - metabolism; Corneal Injuries - pathology; Cyclin E - genetics; Cyclin E - metabolism; Cyclin-dependent kinase 2; Cyclin-Dependent Kinase 2 - genetics; Cyclin-Dependent Kinase 2 - metabolism; E-cadherin; Endothelium; Endothelium - drug effects; Endothelium - injuries; Endothelium - metabolism; Epithelial-Mesenchymal Transition - drug effects; Epithelial-Mesenchymal Transition - genetics; fibroblast growth factor; Fibroblast growth factor 2; Fibroblast Growth Factor 2 - antagonists & inhibitors; Fibroblast Growth Factor 2 - genetics; Fibroblast Growth Factor 2 - metabolism; Fibroblast Growth Factor 2 - pharmacology; Fibronectins - genetics; Fibronectins - metabolism; Gene Expression Regulation; Humans; IL-1β; Interleukin 1; Interleukin-1beta - genetics; Interleukin-1beta - metabolism; Interleukin-1beta - pharmacology; Ions; Mesenchyme; Mice; Mice, Inbred C57BL; Mice, Inbred NOD; Oncogene Proteins - genetics; Oncogene Proteins - metabolism; Signal Transduction; siRNA; Snail Family Transcription Factors - genetics; Snail Family Transcription Factors - metabolism; Snail protein; Tissue Culture Techniques; Vimentin - genetics; Vimentin - metabolism; Western blotting; Zinc Finger E-box-Binding Homeobox 1 - genetics; Zinc Finger E-box-Binding Homeobox 1 - metabolism
• ISSN: 1090-0535; 1090-0535

To determine whether the mouse corneal endothelium enters endothelial to mesenchymal transition (EndoMT) following surgical injury in vivo. The corneal endothelium in anesthetized mice was surgically injured in vivo under direct visualization. The secretion of interleukin-1 beta (IL-1β) and fibroblast growth factor 2 (FGF2) into the aqueous humor was analyzed with western blotting. The expression of , , , , , , , , , and was analyzed with semiquantitative RT-PCR in the mouse corneal endothelium ex vivo and in vivo. Knockdown of FGF2 was done using siRNA. was used as a corneal endothelial marker, and Keratocan ( ) was used as a stromal marker. β actin was used as a loading control. Sequential expression of IL-1β and FGF2 was detected in the aqueous humor after surgical injury. FGF2 treatment induced expression of endothelial to mesenchymal transition-related genes including , and in the mouse ex vivo corneal endothelium. This led to increased expression of , , , and and suppression of in a time-dependent manner. Expression of , , , , , , , , and was completely abolished by FGF2 siRNA knockdown in the mouse corneal endothelium ex vivo. Surgical injury induced expression in the in vivo mouse corneal endothelium. The injury-dependent expression of , , , , , , , , and and the suppression of were inhibited by siRNA knockdown of FGF in the mouse corneal endothelium in vivo. Moreover, siRNA knockdown of FGF2 inhibited the formation of the injury-dependent retrocorneal membrane in the in vivo mouse corneal endothelium. These findings suggest that after surgical injury, FGF2 induces the expression of EndoMT-related genes , , , , , , , and in the mouse corneal endothelium in vivo, similar to the human corneal endothelium ex vivo.