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Single-Cell Biophysics: Measurement, Modulation, and Modeling

Poster Abstracts

86 

83-POS

Board 42

Traumatic Retraction of Neurons Regulated by Cell-Extracellular Matrix Adhesion

Xueying Shao

, Kwan Kei Wong, Yuan Lin.

The University of Hong Kong, Hong Kong, Hong Kong.

Damage to neural cells and retraction of axons during traumatic brain injury (TBI) are believed

to trigger disintegration of the neural network and eventually lead to severe symptoms such as

permanent memory loss and emotional disturbances. Unfortunately, a method allowing us to

quantitatively characterize the traumatic retraction of neural cells as well identify key players

involved is still lacking. In this study, we used a sharp atomic force microscope (AFM) probe to

transect axons and trigger their retraction while, at the same time, utilizing a total internal

reflection fluorescence microscope (TIRFM) to monitor alterations in the axon cytoskeleton and

cell-ECM adhesion. Interestingly, it was observed that cortical neuron cells cultured on a poly-L-

lysine (PLL) coated coverslip for 3 days retracted much faster than those cultured for 7 days,

presumably because that stronger cell-substrate adhesion was formed after 7 days of culturing.

Furthermore, a well-developed axon may not fully shrink back to the main cell body after the

first axotomy. Instead, the retracting motion can be arrested/stopped. However, axon retraction

will be re-triggered if a second transection is conducted. Finally, we showed that, in general, the

retraction process can be divided into three stages: i) an initial stage with a high retracting speed

of ~0.01 µm/s; ii) an adhesion-dominated stage with a shrinking velocity that is order of

magnitude lower than the initial stage; and iii) the steady-state stage where no change in the axon

length can be detected.