Sonic Hedgehog Expression and Its Role in Form-Deprivation Myopia in Mice

Abstract

Purpose: To investigate whether sonic hedgehog (Shh) plays a role in postnatal eye development and the development of experimentally induced myopia. Methods: Expression of Shh, Patched-1 (Ptc-1), and Gli3 was evaluated in the eyes of 13- to 14-day-old C57B/L6 mice with form-deprivation myopia (FDM) (n = 100) and controls (n = 100) using real-time PCR and Western blot analysis. In a second experiment, 336 mice were divided into two groups: the first wore a unilateral translucent diffuser to induce myopia and the second served as a control. Both groups received four intravitreal injections of either Shh-N (Sonic hedgehog amino-terminal peptide) or cyclopamine (a specific inhibitor of the Shh pathway) every other day. Retinoscopic refraction and axial length measurements were performed on the 11th day of form deprivation. Sections of the eyes were observed using a light microscope. Results: Inducing myopia caused a significant increase in expression of Shh mRNA (7 days: t = 6.09, p = 0.004; 14 days: t = 3.48, p = 0.025) and protein (7 days: t = 4.06, p = 0.015; 14 days: t = 4.25, p = 0.013). Expression of both Gli3 mRNA (t = 7.61, p = 0.002) and protein (t = 2.89, p = 0.045) increased after 7 days of occlusion. Administration of Shh-N stimulated the development of myopia and axial growth in both occluded (refraction: F = 7.49, p = 0.001; axial length: F = 9.89, p < 0.001) and non-occluded eyes (refraction: F = 14.20, p < 0.001; axial length: F = 20.37, p < 0.001). Cyclopamine reduced myopic refractive error and axial elongation in occluded eyes (refraction: F = 27.91, p < 0.001; axial length: F = 15.18, p < 0.001). It also influenced non-occluded eyes, reducing axial growth and shifting the refraction toward hyperopia (refraction: F = 14.81, p < 0.001; axial length: F = 3.99, p = 0.024). No difference in retinal thickness was found between experimental and control eyes. Conclusions: The Shh signaling pathway may influence both form-deprivation myopia and the postnatal growth of eyes with normal visual input.