Expression of the inhibitory receptor subunit GABAA ?2 in neurons and glia of zebrafish embryos

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Issue Date
2015-04-10
Authors
Vazquez, Jose L.
Andera, Katelynn M.
Bryant, MacKenzie M.
Fuerch, Derek J.
Kelly, Brianna D.
Marte, Victoria C.
Miller, Ilsa S.
Murray, Kayla M.
Ryan, Caitlin M.
Turkewitz, Daniel R.
Publisher
Keywords
Neurons , Glia , GABA , Neuroscience , Biology , Zebrafish , Epilepsy , Fluorescent , RNA , Antibody
Abstract
The inhibitory GABA_A receptors regulate most regions of the nervous system. Identification of the cell types and tissues that express different GABA_A receptor subunits could greatly benefit research into a variety of mental illnesses including epilepsy, drug abuse, anxiety disorders, depression, and schizophrenia. As part of a biotechniques course at SUNY Alfred State College of Technology, students identified regions in the nervous system of zebrafish embryos that expressed the GABA_A ?2 subunit. Students synthesized fluorescent mRNA in situ probes directed against the GABA_A ?2 subunit using a variety of technologies. First, students cloned a fragment of the GABA_A ?2 subunit gene from whole RNA using reverse transcription PCR. They then cloned the resulting cDNA into a plasmid vector. Utilizing this plasmid, students generated fluorescent in situ probes. Combining these in situ probes with antibodies specific to neurons and glia allowed students to determine which cell types express the GABA_A ?2 subunit. This project allowed students to carry out a modern biotechnology project from design to completion. Students generated previously unpublished data and produced stunning images while gaining practical skills applicable to both academic and industry research projects.
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