TY  - THES
AU  - Da Silva Santos, Telma
TI  - Axon growth of CNS neurons in three dimensions is amoeboid-like and independent of adhesions
PB  - Rheinische Friedrich-Wilhelms-Universität Bonn
VL  - Dissertation
CY  - Bonn
M1  - DZNE-2020-00275
SP  - 137 pages : 39 figures, 12 movies, CD
PY  - 2019
N1  - Dissertation, Rheinische Friedrich-Wilhelms-Universität Bonn, 2019
AB  - Neurons physiologically grow in a three-dimensional (3D) environment. However, conventional cell culture systems for neuronal studies have been made on a hard and flat surface. Dimensionality is known to affect cytoskeletal organization, molecular pathways and cell dynamics in fibroblasts, dendritic cells and cancer cell lines. Here, I investigated axonal growth in vitro through a reproducible method representing a 3D and more physiological system. Overall neuronal growth is higher in 3D compared to the conventional two dimensions (2D) cultures, and the growth pattern matches what I observed in a more physiological set-up, ex vivo. The axonal growth cone morphology differs extensively, suggesting that artifacts arise from the mechanical properties of the 2D in vitro systems. Thus, the alterations on the actin cytoskeleton that I measured contradict the clutch model hypothesis to explain growth cone advancement. Furthermore, I provide pieces of evidence of the forces involved in physiological axonal elongation. I demonstrate that growth cone advancement is independent of adhesions, in accordance with the amoeboid movement, which is intimately linked with actin dynamics and myosin II activity. ln this work, I demonstrate how a 3D cell culture system offers insights on neuronal physiological growth and the artifacts associated with the conventional culture methods in 2D. Thereby, this work tightens the gap between in vitro andin vivo research.One Sentence Summary:The physiological motility of the axonal growth cone is amoeboid and independent of adhesions.
LB  - PUB:(DE-HGF)11
UR  - https://pub.dzne.de/record/144833
ER  -