Previously, our lab characterized the regeneration of zebrafish rod and cone photoreceptors in response to light-induced damage.  To determine the breadth of cell types that can regenerate within the retina and characterize the underlying mechanisms involved in neuronal regeneration and cellular differentiation, I intravitreally injected ouabain, a sodium-potassium ATPase inhibitor, into the zebrafish eye and determined that ouabain destroys the inner neurons of the zebrafish retina, which is in contrast to the light-induced photoreceptor cell death in the outer retina.

Histological analysis demonstrated that ouabain (2,0 µM) disrupted the ganglion cell layer (GCL), inner nuclear layer (INL), and both plexiform layers (IPL and OPL) within three days after injection, while leaving photoreceptors intact.  Immunohistochemical analysis with an antibody that detects proliferating cell nuclear antigen (PCNA), which labels cells undergoing DNA replication and/or repair, was used to characterize the large proliferative response initiated by the Müller glia at 3 dpi.  Retinal sections labeled with HuC/D, an antibody used to label differentiated amacrine and ganglion cells, characterized the subsequent regeneration of two classes of known inner retinal cell types within sixty days post ouabain injection.  Taken together, these data suggest that retinal regeneration is a highly coordinated event that is largely dependent on the types of cells destroyed and the level of destruction sustained by the retina.  Currently, I am interested in the activation profile of microglial/macrophage cells that infiltrate and proliferate during the early retinal damage.

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