To Study The Effect Of α-Syn Oligomer On NMDAR On Neuronal Membrane Surface

Objective: To study the effect of α-synuclein(α-Syn)oligomer on the viability of neurons and N-methyl-D-aspartic acid receptor(NMDAR)on the membrane surface of neurons.Methods: Incubation of neurons by extracellular addition α-Syn oligomer.Western blot and immunofluorescence were used to detect whether the extracellular α-Syn oligomers could enter neurons and have an effect on the expression of NMDAR on the surface of neurons.The brains of healthy newborn SD rats(born within 24 h)were dissected under strict aseptic condition,and the primary neurons were extracted and cultured from the brains.The α-Syn oligomers were prepared by PAGE gel protein micro recovery method,and the neurons were incubated with extracellular α-Syn oligomers.The effects of extracellular α-Syn oligomers on the viability of neurons were detected by CCK-8,immunofluorescence and Western blot were used to detect the effect of α-Syn oligomer on the expression of NMDAR.Results: 1.The Western blot suggests that the incubation protein was α-Syn oligomer.2.The results of CCK-8 showed that the activity of neurons in the experimental group with α-Syn oligomer was lower than that in the control group without α-Syn oligomer.3.The results of Western blot and immunofluorescence showed that there were more α-Syn oligomers detected in the neurons of the experimental group(α-Syn 1 group,α-Syn 2 group)with α-Syn oligomer than that of the control group(control group)without α-Syn oligomer,and the α-Syn 2 group(2 μg/ml with α-Syn oligomer)was more than that of the α-Syn 1 group(1 μg/ml with α-Syn oligomer).4.The results of Western blot and immunofluorescence showed that α-Syn oligomers increased in the experimental group(α-Syn 1,α-Syn 2)compared with the control group(control group);NR1 decreased in the membrane protein of neurons,increased in the cytoplasmic protein of neurons,remained unchanged in the total protein of neurons,and the change of NMDAR in α-Syn 2 group(2 μg/ml with α-Syn oligomer)was more significant than that in α-Syn 1 group(1 μg/ml with α-syn oligomer).Conclusions: 1.The addition of α-Syn oligomer in vitro can reduce the viability of neurons.2.The α-Syn oligomers can enter neurons.3.When α-syn oligomer enters neurons,it can affect the NMDAR on the surface of neuron membrane and promote the internalization of NMDAR on the surface of neuron membrane.