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The coordination of netrin signal transduction via TUBB3 and JNK1 in axon guidance
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The coordination of netrin signal transduction via TUBB3 and JNK1 in axon guidance
Contents
Abstract
Ackownledgements
Contents
List of Figures
List of Abbreviations
1 Background
1.1 The nervous system
1.2 Axon guidance
1.3 Netrins
1.4 Netrin-1 receptors
1.4.1 Deleted in colorectal cancer (DCC)
1.4.2 Down syndrome cell adhesion molecule (DSCAM)
1.4.3 Uncoordinated 5 (UNC5)
1.5 Netrin-1 signaling
1.6 Cytoskeleton
1.6.1 Microtubule
1.6.2 Actin
1.7 Tubulin Beta-3 chain (TUBB3)
1.8 c-Jun N-terminal kinase 1 (JNK1)
2 Significance
3 Material and methods
3.1 Plasmids and oligonucleotides
3.2 Antibodies and other reagents
3.3 Immunoprecipitation and western blot analysis
3.4 Dissociated primary neuron culture and RNAi electroporation
3.5 Immunocytochemistry
3.6 Protein purification
3.7 Microtubule cosedimentation assay
3.8 Axon outgrowth assay
3.9 Chick spinal cord axon turning assay
3.10 Analysis of commissural axon projection in vivo
3.11 JNK activity assay
4 Results
4.1 TUBB3 interacts with DCC
4.2 MT dynamics modulates the interaction of TUBB3 with DCC
4.2.1 The netrin-1 induction of TUBB3/DCC interaction requires MT dynamics
4.2.2 The activity of Src family kinases is required for netrin-1 induction of TUBB3 tyrosine phosphorylation and the interaction of TUBB3 with DCC
4.3 TUBB3 is required for netrin-1-induced neurite outgrowth
4.3.1 I The knockdown of TUBB3 inhibits netrin-1-induced axon outgrowth in cortical neurons
4.3.2 TUBB3 does not bind to endogenous TrkB, a BDNF receptor, and is not involved in BDNF-promoted neurite outgrowth
4.3.3 TUBB3 knockdown inhibits netrin-1-induced axon outgrowth of chick dorsal spinal cord explants
4.4 TUBB3 is required for axon attraction by netrin-1
4.4.1 TUBB3 is required for netrin-1 attraction of spinal commissural axons
4.4.2 The intracellular P2-3 domain of DCC inhibits the interaction of TUBB3 with full-length wild-type DCC, as well as netrin-1-induced neurite outgrowth and commissural axon attraction
4.5 TUBB3 is required for spinal commissural axon projection in vivo
4.5.1 TUBB3 knockdown inhibits of commissural axon projection in vivo
4.5.2 TUBB3 is essential for spinal cord commissural axon pathfinding in vivo
4.6 Netrin-1 increases JNK1 activity
4.7 Netrin-1-induced JNK1 activation requires the intracellular domains of DCC and DSCAM and is inhibit by JNK inhibitors
4.8 Netrin-1 induces endogenous JNK activity
4.9 JNK activation in the developing spinal cord
4.10 JNK1 inhibition blocks netrin-1-induced neurite outgrowth
4.11 JNK1 is required for netrin-1-mediated commissural axon attraction
4.12 JNK1 is essential for spinal cord commissural axon projection and pathfinding in vivo
5 Discussion
5.1 TUBB3 is an essential signaling component in netrin-1-mediated axon outgrowth/guidance
5.2 Netrin/DCC signaling couples directly to MT dynamics
5.3 JNK1 is required for commissural axon projection and pathfinding in the developing spinal cord
5.4 JNK1 coordinates netrin/DCC, netrin/UNC5 and netrin/DSCAM signaling
5.5 Coordination of MT/actin dynamics and JNK1 in axon guidance
ReferenceBài viết liên quan