Lab tour

The Huganir lab is located on the 10th floor of Hunterian building. In the lab, we use a combinatino of molecular, biochemical, cellular, electrophysiological, and optical imaging techniques to study glutamate receptor trafficking, synaptic plasticity and the implication of glutamate receptor trafficking in learning and memory.

Imaging AMPAR Insertion using total internal reflection fluorescent microscopy (TIR-FM)

Insertion of AMPAR to neuronal surface is one of the key steps in synaptic delivery of AMPARs. To directly dissecting the molecular mechanisms governing AMPAR insertion, I have set up a custom TIRF microscope to directly visualize individual insertion events of AMPAR subunit GluR1. This TIRF system was based on a Zeiss AxioObserver and a Zeiss TIRF slider. A Coherent Sapphire 488 -50 mW (OEM version) laser was chosen as the excitation source. The excitation laser was coupled to the TIRF slider through a KineFlex optical fiber, and a Uniblitz shutter was used to control laser on/off. Examples below are N-terminal pHluorin tagged GluR1 imaged under TIRF to visualizing insertion events, and morphology of neurons was visualized withe epi-fluorescent imaging. Click Here for reference.

(The TIRF system shares a 6x4 airtable with the long term imaging system)

The setup: (lasers)

The setup: (microscope)

(Visulizing AMPAR Insertion with TIRF imaging,

Magenta: Neuron under epi-illumination; Greem: Insertion observed with TIRF)

(Y-T maximum intensity projection of above AMPAR insertion event to facilitate visualizing insertion events)

(AMPAR Insertion and Diffusion on dendrites, high resolution.

Magenta: dendrite and spines morphology; Green: Insertion of AMPAR )

By imaging pHluorin-GluR1, in combination with other experimental approaches, the following is what we think a mechanism that regulates activity dependent GluR1 insertion:

Questions or Comments, please contact Da-Ting Lin