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

In Japanese