Expression of transcription factors and crystallin proteins during rat lens regeneration

PURPOSE: To establish a model of lens regeneration in rats and to detect the expression of transcription factor and crystallin genes. METHODS: An extracapsular lens extraction (ECLE) was performed in Sprague-Dawley rats. Examinations with slit-lamp and histological analysis were performed at various time points after ECLE. Real-time PCR and/or immunofluorescence were performed to detect the expression of the lens transcription factors paired box 6 (Pax6), prospero homeobox 1 (Prox1), and forkhead box E3 (Foxe3) and α-, β-, and γ-crystallin (Cryaa, Cryab, Crybb1, Crybb2, Cryba2, and Crygd, respectively). RESULTS: Lens epithelial cells (LECs) were left behind under the anterior capsule immediately after ECLE. Lens fiber differentiation had occurred in the peripheral capsular bag in all rats 3 days after ECLE. One month after surgery, all capsular bags were filled with new semitransparent lenticular structures displaying an established equator with well differentiated bow regions. The mRNA-expression quantity of lens transcription factors and α-, β-, and γ- crystallin increased after ECLE. Pax6 was expressed in both LECs and the newly regenerated lens fiber cells, Prox1 was expressed both in LECs and differentiating lens fiber cells, and Foxe3 was confined to LECs. CONCLUSIONS: Lens fiber differentiation during regeneration follows a process similar to embryological development, with proliferation of epithelial cells along the anterior and posterior capsule, elongation of the posterior epithelial cells, and differentiation of epithelial cells into lens fibers. The regenerated lens contains proteins and transcription factors similar to those found in normal lenses. Inductive interactions seen during lens development are not necessary for lens regeneration.