Abstract
Purpose: :
To identify transgenic mouse lines that have different cell types selectively labeled with GFP. These mice would allow dissecting the cellular components of complex retinal circuits.
Methods: :
We analysed retinas from ~500 different mouse lines of the Gene Expression Nervous System Atlas project. Each mouse line expresses GFP from a different promoter in Bacterial Artificial Chromosomes. One retina of each eye pair was processed for wholemount labelling with anti-GFP and anti-ChAT antibodies as well as DAPI. We obtained 3D confocal scans from each GFP positive retinas. Stratification was quantified relative to the ChAT strata. For detailed characterizations of cell types, we sectioned the second retina and stained with a variety of antibodies. Moreover, two photon laser targeted electrophysiological recordings at different developmental stages were performed.
Results: :
We identified ~100 mouse lines with stratum and/or cell type-specific labeling. Among others we identified mouse lines in which the different inhibitory cell types of the night vision and directional selective circuit are selectively labeled. We quantified the stratification patterns to predict potential synaptic connectivity between marked cells of different lines. GFP expression was high enough in many lines to allow two-photon laser targeted electrophysiological recordings in adult as well as in developing retinas. The 3D scans and quantifications will be available from a public database and the mouse lines can be ordered from MMRRC.
Conclusions: :
Our mapping of retinal cell types to specific BAC transgenic mouse lines enables targeted recordings, imaging and gene expression studies of different cell types from developing or mature retinal circuits.
Keywords: retina: proximal (bipolar, amacrine, and ganglion cells) • retina: distal (photoreceptors, horizontal cells, bipolar cells) • gene/expression