RT Journal Article
SR Electronic
T1 Biology and Bias in Cell Type-Specific RNAseq of Nucleus Accumbens Medium Spiny Neurons
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 444315
DO 10.1101/444315
A1 Hope Kronman
A1 Felix Richter
A1 Benoit Labonté
A1 Ramesh Chandra
A1 Shan Zhao
A1 Gabriel Hoffman
A1 Mary Kay Lobo
A1 Eric E. Schadt
A1 Eric J. Nestler
YR 2018
UL http://biorxiv.org/content/early/2018/10/16/444315.abstract
AB Isolation of cell populations is untangling complex biological interactions, but studies comparing methodologies lack in vivo complexity and draw limited conclusions about the types of transcripts identified by each technique. Furthermore, few studies compare FACS-based techniques to ribosomal affinity purification, and none do so genome-wide. We addressed this gap by systematically comparing nuclear-FACS, whole cell-FACS, and RiboTag affinity purification in the context of D1 or D2 dopamine receptor-expressing medium spiny neuron (MSN) subtypes of the nucleus accumbens (NAc), a key brain reward region. We find that nuclear-FACS-seq generates a substantially longer list of differentially expressed genes between these cell types, and a significantly larger number of neuropsychiatric GWAS hits than the other two methods. RiboTag-seq has much lower coverage of the transcriptome than the other methods, but very efficiently distinguishes D1- and D2-MSNs. We also demonstrate differences between D1- and D2-MSNs with respect to RNA localization, suggesting fundamental cell type differences in mechanisms of transcriptional regulation and subcellular transport of RNAs. Together, these findings guide the field in selecting the RNAseq method that best suits the scientific questions under investigation.