Single-neuron transcriptome and methylome sequencing for epigenomic analysis of aging.
Enormous heterogeneity in transcription and signaling is the feature that slows down progress in our understanding of the mechanisms of normal aging and age-related diseases. This is critical for neurobiology of aging where the enormous diversity of neuronal populations presents a significant challenge in experimental design. Here, we introduce Aplysia as a model for genomic analysis of aging at the single-cell level and provide protocols for integrated transcriptome and methylome profiling of individually identified neurons during the aging process. These single-cell RNA-seq and DNA methylation assays (methyl-capture/methyl enrichment) are compatible with all major next generation sequencing platforms (we used Roche/454 and SOLiD/Life Technologies as illustrative examples) and can be used to integrate an epigenetic signature with transcriptional output. The described sequencing library construction protocol provides both quantitative and directional information from transcriptional profiling of individual cells. Our results also confirm that different copies of DNA in polyploid Aplysia neurons behave similarly with respect to their DNA methylation.
The Whitney Laboratory for Marine Biosciences, University of Florida, Saint Augustine, FL, USA.
Leonid L
LL
Moroz
Andrea B
AB
Kohn
D000375:Aging / Q000235:genetics*; D000818:Animals; D001048:Aplysia / Q000235:genetics; D001483:Base Sequence; D019175:DNA Methylation / Q000235:genetics*; D057890:Epigenomics / Q000379:methods*; D020869:Gene Expression Profiling; D059014:High-Throughput Nucleotide Sequencing; D009474:Neurons / Q000166:cytology; D017423:Sequence Analysis, RNA / Q000379:methods*; D059467:Transcriptome / Q000235:genetics*