The inhibitory GABAA receptors regulate most regions of the nervous system. Identification of the cell types and tissues that express different GABAA 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 GABAA α2 subunit. Students synthesized fluorescent mRNA in situ probes directed against the GABAA α2 subunit using a variety of technologies. First, students cloned a fragment of the GABAA α2 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 GABAA α2 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 Students generated previously unpublished data and produced stunning images while gaining practical skills applicable to both academic and industry research projects
