Purpose Pseudorabies virus (PRV) constructed to express an enhanced green fluorescent protein (EGFP) was injected into vitreous body to label homolateral and contralateral ipRGCs and neurons in SCN, so to provide an important tool for exploring morphological changes of ipRGCs in retinal degeneration.
Methods Unilateral PRV injected into the vitreous body of the Long Evans rats and RCS rats eyes to transsynaptically infect a restricted set of retinal ganglion cells (ipRGCs) and retinorecipient neurons including neurons in the hypothalamic suprachiasmatic nucleus (SCN).
Results At longer post-injection times, retinal ganglion cells in the contralateral eye also expressed EGFP, becoming infected after transsynaptic uptake and retrograde transport from infected retinorecipient neurons. Retinal ganglion cells that expressed EGFP were easily identified in retinal whole mounts viewed under epifluorescence. Whole-cell patch-clamp recordings revealed that the physiological properties of PRV-infected RGCs were within the range of properties observed in non-infected RGCs neurons.
Conclusion The results suggest that PRV is a powerful tool for the transsynaptic labeling of ipRGCs, so to be identified in vitro by their expression of EGFP for analyzing electrophysiological and morphological details. |