| Abstract |
Participants will be able to describe single cell analysis to broadly classify cell types and observe differences in chromatin accessibility and cell type abundance between treatment groups. Choline plays integral roles in neurodevelopment through the production of acetylcholine, formation of phosphatidyl choline, and as the primary dietary methyl donor. Despite the increased need for choline during pregnancy, 90% of women do not consume the Adequate Intake level, and choline is not currently included in standard prenatal vitamins. Maternal choline supplementation (MCS) has been shown to improve cognitive functioning and lessen neurodegeneration. The mechanism(s) underlying these benefits, however, are largely unknown. The purpose of this pilot study was to evaluate the effectiveness of nuclei extraction from frozen tissues and to ensure that we can produce high quality libraries for analysis of MCS and behavior outcomes. Using methods developed by the Shendure lab, cortical tissues from a subset (n=3/group) of behaviorally tested mice in an MCS research study were used to create snATAC-seq libraries. Cells were classified into 6 broad cell-type categories by cluster location on UMAPs, marker gene accessibility levels, and Gene Ontology (GO) functional pathways. Quality control thresholds from TSS enrichment and unique barcode reads per cell confirm high quality cells and great libraries have been created. We broadly classified 24 clusters containing 20,596 cells as excitatory neurons (6), interneurons (3), oligodendrocytes (5), astrocytes (3), microglia (2), or endothelial cells (2). Despite the small sample size, effects in cell number and chromatin accessibility have been detected. The quality control metrics from our pilot study provide confidence in the utility of archived brain tissues to assess the potential correlations between MCS and behavioral outcomes. |
, Bo Shui , Barbara J. Strupp , Jones Keith 
