ON-bipolar cell gene expression during retinal degeneration: Implications for optogenetic visual restoration

• DOI: 10.1016/j.exer.2021.108553
• Publication Date: 2021-03-31T19:43:16Z
• Authors: Michael J. Gilhooley; Doron G. Hickey; Moritz Lindner; Teele Palumaa; Steven Hughes; Stuart N. Peirson; Robert E. MacLaren; Mark W. Hankins
• Keywords: 0301 basic medicine; Retinal Bipolar Cells; Genetic Vectors; Retinal Degeneration; Mice, Transgenic; Inherited retinal degeneration; Dependovirus; Flow Cytometry; Real-Time Polymerase Chain Reaction; Immunohistochemistry; Spermidine Synthase; Article; 3. Good health; Optogenetics; Mice; 03 medical and health sciences; Gene Expression Regulation; Contactin 1; Animals; Annexin A7; Female; Gene expression; Bipolar cell; Sulfatases

Retinal bipolar cells survive even in the later stages of inherited retinal degenerations (IRDs) and so are attractive targets for optogenetic approaches to vision restoration. However, it is not known what extent remodelling that these undergo during degeneration affects their function. Specifically, unclear if they free from metabolic stress, receptive adeno-associated viral vectors, suitable opsin-based tools able propagate signals by releasing neurotransmitter. Fluorescence activated cell sorting (FACS) was performed isolate labelled dissociated retinae litter-mates with or without IRD mutation Pde6brd1/rd1 selectively expressing an enhanced yellow fluorescent protein (EYFP) as a marker ON-bipolar cells. Subsequent mRNA extraction allowed Illumina® microarray comparison gene expression degenerate those wild type retinae. Changes four candidate genes were further investigated at level using immunohistochemistry over course degeneration. A total sixty differentially expressed transcripts reached statistical significance: did include any directly associated native primary signalling, nor changes consistent stress. Four significantly altered (Srm2, Slf2, Anxa7 & Cntn1), implicated synaptic remodelling, neurotransmitter release vector entry had immunohistochemical staining colocalising markers varying Our findings suggest relatively few context degeneration: despite likely remain viable In addition, several where seen could provide basis investigations enhance efficacy therapies.