A Cytomic Approach Towards Genomic Individuality Of Neurons

Here, we describe an approach for the DNA quantification of single cells in brain slices based on image cytometry (IC) that allows mapping the distribution of neurons with DNA content variation (DCV) in the context of preserved tissue architecture. The method had been optimized for DNA quantification of identified neurons but could easily be adapted to other tissues. It had been validated against chromogenic in situ hybridization (CISH) with chromosome-specific probes and laser microdissection followed by quantitative PCR (qPCR) of alu repeats. It can be combined with immunocytochemical detection of specific marker proteins which allow for further specification of cellular identity in the context of defined brain pathology. The method can be applied in a high-throughput mode where it allows analyzing 500,000 neurons per brain in a reasonable time. The combination of cytometry with molecular biological characterization of single microscopically identified neurons as outlined here might be a promising approach to study molecular individuality of neurons in the context of its physiological or pathophysiological environment. It reflects the concept of cytomics and will forward our understanding of the molecular architecture and functionality of neuronal systems.