Session: 7

PS7-09 | Multiple states of neuronal development in the adult hippocampus revealed by single-nuclei RNA-seq

Natalí Rasetto

Laboratorio Plasticidad Neuronal

Adult hippocampal neurogenesis plays a critical role in spatial memory formation and retrieval, context discrimination, and clearance of memory traces. In the mouse dentate gyrus, the maturation of adult-born granule cells (GCs) lasts several weeks and can be divided in 4 phases based on electrophysiological and morphological features. However, the molecular mechanisms that control the progression through those discrete phases are still unknown. We have proposed that such progression is driven by sequential changes in the program of gene expression, and should be revealed by transcriptome analysis. We thus set up an approach for high-throughput single-nuclei RNA sequencing applying Chromium 10X Genomics technology to interrogate the transcriptomic composition of new GCs at 1, 2, 4 and 8 weeks of age. We used double transgenic mice, Ascl1CreERT2;CAGfloxStopSun1sfGFP to allow conditional expression of Sun-1/sfGFP in the nuclear membrane of GCs at identified ages. Fluorescent nuclei were purified using FACS. We have obtained a dataset containing 18,000 distinct transcripts in 16,000 nuclei. Preliminary bioinformatic analysis resulted in multiple clusters corresponding to different stages of neuronal differentiation. Interestingly, radial glial cells move along a pathway of >12 transitions to reach a mature neuronal phenotype. We are currently identifying specific markers to discriminate possible functional differences among those clusters.