Date of Completion
Physiology and Neurobiology
Molecular and Cellular Neuroscience | Molecular Genetics | Nervous System | Nervous System Diseases | Neurology | Neurosciences
Neurons are a post-mitotic cell population, and therefore, they are not able to regenerate in vivo after a traumatic injury. Because inhibitory GABAergic interneurons and oligodendrocyte precursor cells (OPCs) are derived from the same precursor, recent studies have focused on transforming these OPCs into GABAergic neurons. However, there are different types of GABAergic interneurons that have different electrophysiological responses, which can lead to functional differences. The Nishiyama laboratory had already used a key gene in GABAergic interneuron and OPC differentiation, Distal-less homeobox 2 (Dlx-2), to transfect OPCs; early electrophysiology tests showed most of these transfected cells behaved like immature neurons, but some behaved how fast-spiking parvalbumin-containing GABAergic neurons would. To further clarify what type of GABAergic interneurons our Dlx-2 transfected cells are becoming, I chose fourteen genes to quantify and compare, utilizing reverse-transcriptase polymerase chain reactions (qRT-PCR). After designing and optimizing primer pairs to target these genes, five replicates of transfected cells were tested; results were analyzed using both the 2−ΔΔCT method and by evaluating raw cycle thresholds for each gene. Gad67, the gene responsible for producing GABA, was significantly more expressed in the Dlx-2-transfected samples, showing that these cells are differentiating into GABAergic interneurons. Although a few genes linked to the parvalbumin- and somatostatin-containing types of GABAergic interneurons were significantly more expressed in the Dlx-2-transfected samples, most genes were not significantly changed, suggesting that these cells are either expressing a mixed phenotype or are not developed enough at this time point to clearly show a subtype.
Nolan, Timothy, "Quantifying Expression of Interneuron Subtype Markers for Dlx-2 Transfected NG2 Cells" (2019). Honors Scholar Theses. 647.