Birthdating studies

The vertebrate retina uses diverse neuronal cell types arrayed into complex neural circuits to extract, process, and relay information from the visual scene to the higher order processing centers of the brain.Amacrine cells, a class of interneurons, are thought to mediate much of the processing of the visual signal that occurs within the retina.Although amacrine cells display extensive morphological diversity, the molecular nature of this diversity is largely unknown.Furthermore, it is not known how this diversity arises during development.Here, we have combined in vivo genetic labeling, single cell genome-wide expression profiling, and classical birthdating to (The vertebrate retina is an excellent system for studying neuronal diversity and how this diversity arises during development.The retina contains 5 major neuronal cell classes and 1 glial cell type.

Amacrine cell types also can be characterized based on their expression of a limited set of molecular markers; however, there are far fewer molecular markers than known morphological types.This paucity of markers is a serious impediment to characterization of this important cell class (3, 4).To characterize the diversity of amacrine cells and how it arises during development, we first used genetic reporters to label individual amacrine cells within the developing retina and observed a range of distinct morphologies.We then used microarray-based expression profiling of single cells to analyze the distinct transcriptional programs of amacrine cells during development.Single cell profiling allowed us to investigate the extent of molecular diversity within the amacrine cell class and led to the identification of previously unknown molecular markers for known and previously uncharacterized amacrine cells, as well as a classification of single amacrine cells according to transcriptional identity.We also unexpectedly observed that GABAergic amacrine cells emerge before glycinergic amacrine cells in development.

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