The Rho-GTPases are intracellular signaling molecules that regulate actin dynamics, cell-cycle progression, cell adhesion, and gene transcription. Signal transduction mediated by RhoA, a well characterized member of the Rho family, was found to be responsible for the cell contraction observed in the early phase of apoptosis. Activation of RhoA and its downstream effector Rho-associated-coiled-coil-kinase (ROCK), an event stimulated by repulsive axon guidance cues during development, is also likely to be responsible for the collapse of the neurite network, another early feature of neuronal apoptosis. Inhibition of the RhoA-ROCK signaling pathway therefore emerges as an attractive strategy to promote cell survival and axon regeneration after central nervous system (CNS) injury or in neurodegenerative diseases. Here, we aim to analyze the influence of RhoA-ROCK signaling pathway on retinal cell survival. Our approach is to use the commercially available ROCK inhibitors H-1152P, HA1077, and Y-27632 in mouse retinal explant cultures and to determine the extent of cell damage by ethidium homodimer-1 and DAPI staining. We combine these in vitro experiments with immunohistochemistry and western blot to characterize certain intra- and extracellular changes at the protein level associated with ROCK inhibition. Our preliminary results showed a neuroprotective effect of H-1152P on the ganglion cell layer, which was associated with a reduction in glial cell reactivity and the release of various cytokines.