Generating dorsal cerebral organoids with GABRA1 mutation to observe neurodevelopment
When studying genetic mutations within the brain, it becomes difficult to accurately observe how a mutation affects neurodevelopment due to lack of models of the human brain. Animal models are inadequate for neuroscience research due to the complexity of the human brain. Cerebral organoid generation is a promising alternative, as these can mimic the basic structural and functional properties of the brain. We cultured induced pluripotent cells (iPSCs) with the GABRA1 mutation and used them to create dorsal forebrain organoids to observe neurodevelopment. On day 15, RNA was extracted from dorsal and ventral organoids for comparison via a reverse transcription RT-qPCR to measure the expression of PAX6. Results showed an upregulation of PAX6 in dorsal GABRA1 organoids, which aligns with current literature. They were also used for cryosectioning and immunofluorescence where beta-tubulin III was used to identify neuronal differentiation. Neuronal formation was observed on day 15 and formation continued to expand on day 25. These results are the foundation of an ongoing experiment to continuously produce cerebral organoids from iPSCs with mutations to study neurological disorders.
