A reliable work ow for single-nuclei RNA- seq of hippocampal adult-born granule cell

RASETTO, NATALI BELEN; BERARDINO, ARIEL; VERCESI, MARIA LUZ; VEGA WAICHMAN, TOMAS; FERELLA, LUCIANA; BROWN, JULIANA; ARLOTTA, PAOLA; CHERNOMORETZ, ARIEL; SCHINDER, ALEJANDRO; GIACOMINI, DAMIANA

Congreso; Congreso SAN 2020; 2020

Soc Argentina Neurociencias

Adult hippocampal neurogenesis plays a critical role in spatial memory, retrieval, contextdiscrimination, and clearance of memory traces. In the mouse dentate gyrus, thematuration of adult-born granule cells (GCs) lasts several weeks and can be divided in 4phases based on their functional characteristics. Although the distinctiveelectrophysiological and morphological features of each stage have already beendescribed, the molecular mechanisms that control the progression through the differentstages towards a mature phenotype are still unknown. To investigate the molecularmechanisms instructing structural and functional maturation in each developmentalstage of adult-born GCs, we set up an ef cient protocol for high-throughput single-nucleiRNA sequencing applying Chromium 10X Genomics technology. We used doubletransgenic mice, Ascl1CreERT2;CAG oxStopSun1sfGFP to allow conditional expressionof Sun-1/sfGFP in the nuclear membrane of adult-born GCs. At speci c time points,dentate gyri were microdissected and uorescent nuclei were isolated and puri ed usingFACS. This approach resulted in the puri cation and successful RNA-seq of 3000 nucleiobtained from 2-week-old immature GCs. We present a preliminary bioinformaticanalysis of these data indicating that the experimental work ow described here is areliable method to explore the molecular principles of neuronal maturation andintegration in the adult brain.