School of Science Division of Life Science 30 microRNA Expression Analysis Supervisor: NGUYEN Tuan Anh / LIFS Student: TAO Kwok Wing / BCB Course: UROP 1000, Summer In order to know biological implications of the point mutation at the 448th codon within the G4 protein, it is a prerequisite to synthesize and purify the mutated protein. The report show the experimental workflow employed to achieve the synthesis and purification of the mutant G4 protein, encompassing the selection of expression host, optimization of induction conditions, and refinement of chromatographic purification strategies. Additionally, this report summarizes the analytical results, including yield, purity, and quality of the final product. Therefore, it can be providing a foundational dataset for future investigations into the molecular and biochemical consequences of the 448th codon mutation. Investigating Synaptic Transmission in Alzheimer's Disease Supervisor: PARK Hyo Keun / LIFS Student: DONGRE Jayad Deepak / BCB Course: UROP 2100, Fall The aim of this research project was to use immunolabeling to investigate the effect of tau phosphorylation on the number of synapses in a mouse neuronal cell. A phospho-mimic of the tau protein named T217E is used to replicate tau phosphorylation while T217A – a non-phospho-mimic of tau - is used as a positive control. Genetic modification is used to insert the T217E, T217A or T217 (wild-type tau) gene into plasmids of the AAV virus that function as delivery vectors, which are then transfected into mouse neuronal cells. Confocal microscopy imaging was conducted to observe the presence of synapses within the neurons. It is expected that the cells treated with T217 (wild-type), T217E and T217A will have differences in number of synapses, due to the amino acid modifications at threonine-217. In AD pathology, hyperphosphorylated tau becomes disengaged from microtubules and aggregates inside the cell. This excess accumulation can form in the synapse too, destroying the structure of the synapse and leading to reduced synaptic transmission and eventual cell death. Thus, it is predicted that cells treated with T217E will resemble AD pathology and have less number of synapses than T217, while cells treated with T217A will show similar synapse quantity as the wild-type. Investigating Synaptic Transmission in Alzheimer's Disease Supervisor: PARK Hyo Keun / LIFS Student: LAI Chun Kan / BCB Course: UROP 1000, Summer Alzheimer’s disease, a type of neurodegenerative disorder first identified in 1907 by Alois Alzheimer, is widely believed to be linked to the disruption of the physiological behavior of tau protein. Mutation of Tau protein including R406W, V337M and P301L alone is sufficient to reduce presynaptic activity which ultimately leads to neurodegenerative disease. In this study, we used rat primary neuron culture and expressed both wildtype (WT) and pathogenic mutant Tau P301L by transient transfection. Upon stimulation, a noticeable defect in synaptic vesicle release was recorded in Tau P301L expressing neurons, suggesting pathogenic mutant Tau protein P301L impedes neurotransmission.
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