Journal: Scientific Reports
Article Title: Cholesterol modulates presynaptic and postsynaptic properties of excitatory synaptic transmission
Figure Lengend Snippet: Cholesterol depletion reduces the fraction of synaptic immobile NMDARs. ( A ) Surface NMDARs were detected using a QD-antibody complex directed against extracellular epitopes in GluN2A or GluN2B. Left, representative summed trajectories of NMDAR-QDs (red) acquired over a period of 25 s (20 Hz frame rate) in hippocampal neurons. Scale bar 5 µm. Right, representative examples of NMDAR reconstructed trajectories. ( B , C ) Diffusion coefficients for synaptic GluN2A-containing NMDARs and GluN2B-containing NMDARs in control and after cholesterol depletion (10 mM MβCD pretreatment, 5 min). ( D , E ) Diffusion coefficients for extrasynaptic GluN2A-containing NMDAR and GluN2B-containing NMDARs in control and after cholesterol depletion. ( F , G ) Diffusion coefficients for the mobile fraction of synaptic GluN2A and GluN2B-containing NMDARs in control and after cholesterol depletion. ( H , I ) Fraction of synaptic immobile receptors in control and after cholesterol depletion. (* p
Article Snippet: Single-particle tracking For endogenous GluN2A or 2B quantum dot (QD) fluorescence particle tracking, hippocampal neurons were incubated with an antibody against the N-terminal extracellular domain of GluN2A (ACG-002 1:500, Alomone Labs) or GluN2B subunits (ACG-003 1:500, Alomone Labs) for 10 min. Neurons were then washed and incubated for 5 min with QD 655 anti-rabbit IgG (Q-11421MP, Invitrogen ).
Techniques: Diffusion-based Assay