Dynamics of presynaptic protein recruitment induced by local presentation of artificial adhesive contacts.
In this study, we introduce a novel approach to induce and observe the formation of presynaptic compartments in axons through a combination of atomic force microscopy (AFM) and fluorescence microscopy. First, we use a poly-D-lysine-coated bead attached to an AFM tip to induce the recruitment of two synaptic proteins, bassoon and synaptophysin, and measure their absolute arrival times to the presynaptic department. We find that bassoon arrives before synaptophysin. Second, we observe the formation of very long (several 10s of μm), structured, protein-containing membranous strings as the AFM tip was withdrawn from the axon. It is conceivable that these strings might be a novel mechanism by which new neurites or branch points along existing neurites may be generated in situ.
Department of Physics, McGill University, Montreal, Canada.
Fernando
F
Suarez
Peter
P
Thostrup
David
D
Colman
Peter
P
Grutter
D000269:Adhesives / Q000494:pharmacology*; D000818:Animals; D001369:Axons / Q000187:drug effects / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology*; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D008863:Microspheres; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D016708:Synaptophysin / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
D000269:Adhesives; C113688:Bsn protein, rat; D009419:Nerve Tissue Proteins; D016708:Synaptophysin; D049452:Green Fluorescent Proteins