Biophysics NeuroScience PNS002
We observed endocytosis of glutamate receptors around the post-synaptic membrane using total internal reflection fluorescence microscopy. Cultured rat neurons were transfected with GluA-SEP, glutamate receptor tagged by pH-sensitive green fluorescent protein, super ecliptic pHluorin (SEP), and PSD95-TagRFPt (a post-synaptic marker). SEP shows fluorescence signals under neutral pH conditions such as extracellular solutions and the lumen of newly formed endocytic vesicles; these signals disappear under acidic conditions. In this video, cell-surface GluA-SEP signals were removed to visualize newly formed vesicles by changing the extracellular pH (7.4) to an acidic pH (6.0) every 4 sec. Images at pH 7.4 and pH 6.0 were acquired alternatively every 2 sec. Chemical stimulation inducing endocytosis was applied at 0 sec. Arrows and arrowheads indicate endocytosis around and out of the post-synaptic membrane, respectively. Scale bar is 5 µm, and the video is played at 132 times normal speed.
|Prefecture||Kyoto||Use of Microscope||Use|
|Region||Yoshida Campus, Kyoto University||Use of Automatic Shooting Devices||Don't Use|
|Shooting Date||2014/6/14||Use of Fluorescent Probe||Use|
|Shooting Time||15:00||Use of Infrared||Don't Use|
|Photographer||Syunpei HUJII||Shooting Interval (Sec.)||4秒|
|Copyright Holder||Syunpei HUJII||Shooting Speed||132倍速|