Spatial-temporal actin dynamics during synaptic plasticity of single dendritic spine investigated by two- photon fluorescence correlation spectroscopy

by Jian Hua Chen
Doctoral thesis
Date of Examination:2013-06-24
Date of issue:2013-12-19
Advisor:Prof. Dr. Peter Jomo Walla
Referee:Prof. Dr. Reinhard Jahn
Referee:Prof. Dr. Andreas Janshoff
Persistent Address: http://dx.doi.org/10.53846/goediss-4291

 

 

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Abstract

English

In the content of my thesis, the construction and calibration of a setup that combined conventional wide field fluorescence microscopy (FM) and two-photon fluorescence correlation spectroscopy (2P-FCS) was first described. The combination of FM and 2P-FCS allowed the investigation of the actin dynamic changes at higher temporal resolution and to gain better insights into the dynamic behavior of the different forms of actin filamentous structures. Samples investigated in the course of this study were living neuronal cells (CA3) that had been transfected with eGFP-actin and fCherry to simultaneously monitor changes in actin dynamics and morphology with the function of Tetraethylammonium (TEA) induced LTP. The actin dynamics before and after TEA stimulation was scrutinized to correlate with the morphological plasticity after long-term potentiation (LTP).  Using FCS measurement, we can distinguish two groups of actin filaments within living cell by two-component fitting models. Further, the absolute particle number and brightness can be compared before and after TEA stimulation. We conclude that dendritic spines underwent morphological enlargement with the function of TEA stimulation are correlated to dynamic changes of actin filaments. These more dynamic actin filaments serve as an important role to drive morphological enlargement after LTP.
Keywords: dendritic spine; two- photon fluorescence correlation spectroscopy; actin dynamics; long term potentiation
 
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