Abstract
Purpose :
Canonical Wnt signaling pathway is an important signaling transduction mechanism involved in essential cellular processes. Optic nerve head (ONH) astrocytes play an important role in glaucoma pathogenesis. The role of Wnt pathway in ONH astrocytes has never been investigated. In this study, we examined the presence of canonical Wnt signaling pathway in ONH astrocytes.
Methods :
Primary mouse optic nerve head (ONH) astrocytes from C57BL/6J mice were cultured and characterized using specific antibodies against glial (GFAP) and S100β. When confluent, cells were serum starved overnight and then treated for 24 hours with 100 nM Wnt3a. Following treatment, cells were collected and cytosolic and nuclear fractions were separated. Expression levels of β-catenin in fractions were determined by western blotting. Images were analyzed via densitometry and fold changes in expression were compared to control group.
Results :
Cells cultured from mouse ONH were positive for GFAP and S100β. Expression levels of β-catenin increased 1.354-fold when treated with 100 nM Wnt3a compared to control in the cytosolic fraction and 1.145-fold when treated with 100 nM Wnt3a compared to control in the nuclear fraction.
Conclusions :
Wnt3a promoted nuclear translocation of β-catenin in mouse ONH astrocytes, which suggests a functional canonical Wnt signaling pathway in mouse ONH. Functions of this pathway in ONH astrocytes will be further studied.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.