Is neuroglobin (Ngb) a perspective for glaucoma?

Abstract

Abstract Purpose To further explore the mechanisms of Ngb function in the retina, and investigate the possible neuroprotective potency of this O2‐binding protein in glaucoma. Our project will potentially open exciting perspectives for the treatment and prognosis of the blinding condition of glaucoma. Methods 1) In vitro: We will set up cultures for primary RGCs. The cultured RGCs will be subjected to hypoxia by incubation in a controlled‐atmosphere incubator. Cell survival and apoptosis will be assessed using MTT method and TUNEL staining. Ngb expression in normoxic and hypoxic cells will be compared. Additionally, the RGCs will be transfected with plasmids expressing Ngb mRNA to evaluate the neuroprotective effect of Ngb. 2) In vivo: The ischemia/reperfusion model and the staurosporine intravitreal injection model cause acute RGC loss. We will also use a laser model to induce chronic elevated IOP by obstructing the aqueous outflow pathway. We will first analyze upregulated Ngb expression in these models. The localization of both Ngb mRNA and protein will be assessed. Ngb‐Tg mice will be subjected to the glaucoma models. We will measure cell density or retinal thickness and perform electroretinography to evaluate the physiological function of retina. We will also analyse the possible contribution of oxidative stress via immunostainings for ROS and determine possible effects on NO scavenging by evaluating the expression of NOS. Furthermore, we will transfect RGCs in vivo using intravitreal injections of a Ngb expressing adeno‐associated virus (AAV) to test the therapeutic potential of Ngb. Conclusion Our project will not only provide us with an improved understanding of neuroglobin, but also open a new perspective for the treatment of retinal ganglion cell apoptosis and glaucomatous neuropathy.