The eyecup preparation has traditionally been used to study retinal physiology in lower vertebrates and in some mammals. The procedures for preparing eyecups of the rat and mouse have not been described, however. We now describe methods for preparing and maintaining viable eyecups for these two species. Eyecups were everted over a plastic dome and held in place between the two halves of a superfusion chamber. Fluid exchange in the chamber was rapid, with near total exchange occurring in 9 s. Eyecup viability was tested by monitoring light-evoked retinal responses as the preparation aged. In both rat and mouse, the amplitude of the electroretinogram (ERG) b-wave decreased slowly, declining to 1/2 maximal amplitude in ≃70 min. Light-evoked spike activity of neurons in the ganglion cell layer remained stable for ≃3 h and attenuated responses were recorded for an additional 1-2 h. Eyecups were able to dark adapt. Retinal sensitivity, tested by monitoring b-wave amplitude, recovered following exposure to an adapting light.
Bibliographical noteFunding Information:
We thank Barry S. Winkler for his suggestions concerning superfusate composition, Paul Ceelen for his excellent technical assistance and Janice I. Gepner and Kathleen R. Zahs for their helpful comments on the manuscript. This work was supported by National Institutes of Health grant EY 04077.
- Electroretinogram (ERG)
- Neuronal activity
- Superfusion chamber