Synaptic Functions of Cell Adhesion and Extracellular Matrix Molecules

Credits: 
1.00
Date: 
30.11.2011
Organizers: 
UNN, IIT
Coordinator: 
Alexander Dityatev
Speaker: 
A. Dityatev, E. Ponimaskin, M. Philippov
Minimal requirements: 
Students are expected to have basic knowledge of cell biology. Experience in in vitro electrophysiology or imaging is an advantage. Good English language skills are essential
Description: 

Synaptic Functions of Cell Adhesion and Extracellular Matrix Molecules

Speakers: Alexander Dityatev, PhD, IIT, Genova, Italy & SUNN, Nizhny Novgorod, Russia; Evgeni Ponimaskin, PhD; MHH, Hannover, Germany; Mikhail Philippov, PhD; SUNN, Nizhny Novgorod, Russia

Course will be held in Conference Hall of the University Fundamental Library ( building №1 of SUNN)

30.11.2011, 14:00-18:00 (4 hours)

01.12.2011, 9:00-18:00 (8 hours)

02.12.2011, 9:00-13:00 (4 hours)

Language: English

Contact person: Susan Asatryan <asatryan@neuro.nnov.ru>

Deadline for registration - 22.11.2011

 Summary:

Processing and storage of information in the CNS relies on formation of synaptic contacts and modification of synaptic efficacy. In addition to neurotransmitters, machinery for their release and neurotransmitter receptors, these processes crucially involve cell adhesion molecules (CAMs) and extracellular matrix (ECM) molecules. These molecules function as “adhesive glue” to stabilize interactions between neurons, as extracellular scaffolds and signal transducers inducing pre- and postsynaptic differentiation, as barriers for diffusion and as long-term modulators of ion channels and neurotransmitter receptors. Mutations in CAMs and ECM molecules lead to severe brain disorders, such as schizophrenia, mental retardation, epilepsy, autism and neurodegeneration. This modular course is (1) to give introduction into functions of most studied CAMs and ECM molecules, (2) to provide methodological details on how to measure their distribution, interactions, posttranslational modifications and signaling and (3) to discuss how the knowledge of molecular mechanisms involved in Alzheimer’s disease can be used for development of therapeutic strategies for treatment of this devastating disease. 

 

Module 1 “Cell adhesion molecules (CAMs) and their functions in the CNS” (A.Dityatev, E. Ponimaskin, 6 hours)

  • Cell-to-cell contacts
  • Homophilic and heterophilic CAMs, cys- and trans-interactions
  • Measurements of adhesive interactions
    • Bead aggregation assay 
    • Laser twizers
    • Atomic force microscopy
  • Cadherins mediate Ca2+ dependent cell adhesion
    • Structure and functions of cadherins
    • Protocadherins and synaptogenesis
  • IgCAMs mediate Ca2+ independent cell adhesion
    • NCAM-mediated intracellular signaling during neurite outgrowth
    • Modulation of cell adhesion by polysialic acid (PSA)
    • PSA and migration of neurons in the rostral migratory stream
    • L1 and axonal growth and guidance
    • P0 and myelin
    • SynCAM and induction of synaptic differentiation
  • Neurexins and neuroligins
  • Ephrins and Eph receptors
  • Diseases associated with human mutations in CAMs
    • L1, CHL1 and mental retardation
    • L1 and fetal alcohol syndrome
    • NCAM and schizophrenia
    • Neuroligins and autism
    • CAM-derived peptides for therapeutic applications
  • Palmitoylation of CAMs as important post-translational fatty acid modification
    • Palmitoylation of NCAM and NCAM-mediated neurite outgrowth
    • Palmitoylation of NCAM and FGF-mediated neurite outgrowth
  • Practical course on palmitoylation measurements
    • Method of metabolic labeling
    • Non-radioactive “click” method
    • Non-radioactive ABE method

Module 2 “Extracellular matrix molecules and their functions in the CNS” (A. Dityatev, 4 hours)

  • Examples of ECM structures
    • Collagen fibrils in skin
    • Basal lamina at the NMJ and other structures
    • Perineuronal nets in the CNS
  • ECM ligands and receptors
    • Integrins and their ligands
    • NG2, CD44 and hyaluronan
  • Outside-in and inside-out signaling
  • Domain structure of ECM components and examples of their functions
    • Laminins and clustering of presynaptic Ca2+ channels
    • Agrin and clustering of acetylcholine receptors
    • Neuronal pentraxins and clustering of AMPA receptors
    • Reelin: formation of the cortex and role in LTP
    • Chondroitin sulfate proteoglycans and inhibition of axonal growth
    • Tenascin-R and the GABAergic interneurons
    • Tenascin-C and Ca2+ channel-dependent synaptic plasticity
    • Thrombospondins and synaptogenesis
  • Assembly of ECM structures
    • Basal lamina of NMJ
    • Perineuronal nets
    • Glial scar
  • Remodeling of ECM and therapeutic applications
    • Chondroitinase ABC improves regeneration
    • Chondroitinase ABC and restoration of developmental plasticity in the cortex
    • Improved regeneration and retarded kindling in tenascin-R deficient mice
  • Extracellular proteases and neuroplasticity 

Module 3 “Applications of fluorescence proteins in neurobiology” (E.Ponimaskin, 2 hours)

  • Introduction in fluorescence proteins (GFP and derivates)
  • The usage of fluorescent proteins for structural and functional studies
  • Analysis of protein distribution
    • Analysis of protein co-localisation
    • Analysis of dynamic distribution by FRAP
    • Analysis of protein-protein interaction by FRET
  • Examples of FRAP, FRET and TIRF applications
  • Small GTPases and their role in regulation of neuronal morphology
  • Introduction to FRET-based biosensors
  • Examples of FRET-based biosensors for RhoA and cdc42

 

Module 4 “Alzheimer’s disease and its therapies” (M. Filippov, 3 hours)

  • Introduction to Alzheimer’s disease
    • Amyloid Plaques and Tau tangles. How to diagnose?
    • Modeling of Alzeheimer disease in mice.
    • Review Amyloid Precursor Protein processing and interactions
      • APP processing (Alfa, Beta and Gamma)
      • A-beta accumulation and its toxicity
      • APP binding partners
      • APP and Apolipoprotein E
      • APP and synaptic vesicles
      • APP and extracellular matrix
        • APP family members and Reelin
        • A-beta and HSPG
    • Key problems:
      • Beta-amyloid plaques destroy the architecture of the nervous system
        • A-beta and neuronal activity
        • Loss of spines and synapses
        • Impairment in learning and memory
        • Accumulation of A-beta around blood vessels and Neuroinflammation
      • Cholinergic deficiency: impairment in NGF transport and cholinergic signaling
      • Neuronal Senescence, Reactive Oxygen Species, BACE1 and APP
        • Cholesterol loss at Neuronal Senescence and allocation of BACE1 to APP
        • BACE1 and its regulation by microRNAs. Why do the microRNA genes are OFF in elders?
      • Gamma secretase. Can we learn something from the genetics?
      • Impaired Extracellular Matrix and A-beta plaques 
  • Therapy
    • Gene therapy approaches
      • Inhibition of BACE
      • Inhibition of neuron specific gamma secretase subunits
      • Endopeptidases and Extracellular Matrix modulating enzymes
    • Immune therapies 
      • “Passive” immune therapy
      • “Active” immune therapy: viral vectors as the antigene presentation tools
    • Other hypothetical possibilities
  • Conclusion

Materials of lectures at zip file

Duration(days): 
3
Place: 
UNN

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