Date of Award

8-1-2015

Degree Type

Dissertation

Degree Name

Doctor of Philosophy

Department

Graduate School of Biomedical Sciences

First Advisor

Raghu R. Krishnamoorthy

Second Advisor

Weiming Mao

Third Advisor

John V. Planz

Abstract

Glaucoma is a group of chronic progressive optic neuropathies commonly characterized by elevated intraocular pressure (IOP) (a subset of glaucoma patients display neurodegenerative effects at ‘normal’ IOP) leading to axonal degeneration, optic nerve head cupping and apoptosis of retinal ganglion cell death (RGCs), which result in visual field defects and blindness. While there are medications available to lower IOP, there is an unmet need for neuroprotective treatments for glaucoma, since some neurodegenerative effects persist despite lowering IOP.

The main focus of this study was on the class 4 POU domain transcription factor, Brn3b, which has been shown to play a key role in the development of RGCs. Two previous studies from other labs showed that a decrease in Brn3b expression occurs in animal model of glaucoma. A recent publication from our laboratory demonstrated neuroprotective effects of adeno-associated virus (AAV) mediated expression of Brn3b in a rat model of ocular hypertension. This research project identified some mechanisms of Brn3b-mediated neuroprotection in cultured PC12 cells (under the condition of hypoxia) and also in vivo in the Morrison’s model of ocular hypertension in rats.

In the first part of the study, we demonstrated the effect of overexpression of Brn3b on various markers of synaptic plasticity in PC12 cells under conditions of normoxia as well as hypoxia. Immunoblot as well as immunocytochemical analyses revealed an increase in expression of neurite growth markers, GAP-43 and ac-TUBA, by Brn3b upregulation both under conditions of normoxia as well as hypoxia. . This suggests that transcription factor Brn3b has the ability to upregulate expression genes contributing to synaptic plasticity genes both under ‘normal’ conditions and during a glaucomatous insult (hypoxia). In the concluding part of this study, cell survival factors including, Bcl-2, Bcl-xL and p-AKT were studied as potential targets of Brn3b-mediated neuroprotection. Adeno-associated virus-mediated expression of Brn3b in rat eyes with elevated IOP promoted an upregulation of Bcl-2, Bcl-xL and p-AKT in RGCs, as determined by immunohistochemistry. Taken together, the evidence suggests that Brn3b has the potential to be developed as a therapeutic agent for neuroprotection during ocular neurodegenerative diseases like glaucoma.

Comments

Available August 2016.