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Updated: Aug 23, 2026
An Isolated Retinal Preparation to Record Light Response from Genetically Labeled Retinal Ganglion Cells
Published on: January 26, 2011
A method for analysis of gene expression in isolated mouse photoreceptor and Müller cells
Karl J Wahlin1, Lynette Lim, Elizabeth A Grice
1Department of Ophthalmology, The Johns Hopkins University School of Medicine, Baltimore, MD 21287-9257, USA.
Purpose:
Molecular analysis of complex phenomena, such as selective death of photoreceptors and their rescue by neuro-protective agents, has been hindered by limitations of techniques for investigating gene expression in individual cells within a heterogeneous tissue such as the retina. The purpose of this study was to develop methods to assess gene expression in single retinal cells.
Methods:
Individual cells from papain-dissociated mouse retinae were captured with micropipettes and identified by morphology and by immunocytochemistry. Single cell cDNA libraries were generated by poly-d(T)-primed reverse transcription, poly-d(A) tailing of first strand cDNA, and en masse PCR-amplification using a custom made oligo-d(T). PCR was used to investigate gene expression in cDNAs from individual cells.
Results:
Dissociated rod and Müller glia cells maintained their morphology, which correlated with their immunocytochemical properties. RPE cells were recognized by their pigmentation. With the exception of bipolar cells, non-photoreceptor neurons were only identifiable by immunocytochemistry. Abundant cDNA could be synthesized from each individual cell. Cell-specific "markers" were detected by PCR almost exclusively in the predicted cell types. The expression of neurotrophic factor receptors was consistent with previous biological studies.
Conclusions:
These studies establish a method to compare, investigate, and analyze gene expression in individual cells of the retina.
