RT Journal Article
SR Electronic
T1 Single-Neuron Gene Expression Analysis Using the Maxwell® 16 LEV System in the Neural Systems and Behavior Course
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 107342
DO 10.1101/107342
A1 Harris, Rayna M.
A1 Otopalik, Adriane G.
A1 Smith, Colin J.
A1 Bucher, Dirk
A1 Golowasch, Jorge
A1 Hofmann, Hans A.
YR 2017
UL http://biorxiv.org/content/early/2017/02/10/107342.abstract
AB Gene expression analysis from single cells has become increasingly prominent across biological disciplines; thus, it is important to train students in these approaches. Here, we present an experimental and analysis pipeline that we developed for the Neural Systems & Behavior (NS&B) course at Marine Biological Laboratory. Our approach used the Maxwell® 16 LEV simplyRNA Tissue Kit and GoTaq® 2-Step RT-qPCR System for gene expression analysis from single neurons of the crustacean stomatogastric ganglion, a model system to study the generation of rhythmic motor patterns. We used double-stranded RNA to knockdown expression of a putative neuromodulator-activated sodium channel. We then examined the electrophysiological responses to known neuromodulators and confirmed that the response was reduced. Finally, we measured how mRNA levels of several ion channel genes changed in response. Our results provide new insights into the neural mechanisms underlying the generation and modulation of rhythmic motor patterns.