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Single-Neuron Gene Expression Analysis Using the Maxwell® 16 LEV System in the Neural Systems and Behavior Course

View ORCID ProfileRayna M. Harris, View ORCID ProfileAdriane G. Otopalik, Colin J. Smith, View ORCID ProfileDirk Bucher, View ORCID ProfileJorge Golowasch, View ORCID ProfileHans A. Hofmann
doi: https://doi.org/10.1101/107342
Rayna M. Harris
1Marine Biological Laboratory;
2The University of Texas at Austin;
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Adriane G. Otopalik
1Marine Biological Laboratory;
3Brandeis University;
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Colin J. Smith
1Marine Biological Laboratory;
4Children’s Hospital of Philadelphia;
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Dirk Bucher
1Marine Biological Laboratory;
5New Jersey Institute of Technology
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Jorge Golowasch
1Marine Biological Laboratory;
5New Jersey Institute of Technology
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Hans A. Hofmann
1Marine Biological Laboratory;
2The University of Texas at Austin;
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ABSTRACT

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.

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Posted February 10, 2017.
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Single-Neuron Gene Expression Analysis Using the Maxwell® 16 LEV System in the Neural Systems and Behavior Course
Rayna M. Harris, Adriane G. Otopalik, Colin J. Smith, Dirk Bucher, Jorge Golowasch, Hans A. Hofmann
bioRxiv 107342; doi: https://doi.org/10.1101/107342
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Single-Neuron Gene Expression Analysis Using the Maxwell® 16 LEV System in the Neural Systems and Behavior Course
Rayna M. Harris, Adriane G. Otopalik, Colin J. Smith, Dirk Bucher, Jorge Golowasch, Hans A. Hofmann
bioRxiv 107342; doi: https://doi.org/10.1101/107342

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