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Spatiotemporal dynamics of transport in the squid giant axon : competition between cargo motor receptors, JIP-1, APP-C, and negative charge

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Please use this identifier to cite or link to this item: http://hdl.handle.net/1928/17453

Spatiotemporal dynamics of transport in the squid giant axon : competition between cargo motor receptors, JIP-1, APP-C, and negative charge

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dc.contributor.author Seamster, Pamela
dc.date.accessioned 2012-02-01T17:38:19Z
dc.date.available 2013-12-31T11:00:07Z
dc.date.issued 2012-02-01
dc.date.submitted December 2011
dc.identifier.uri http://hdl.handle.net/1928/17453
dc.description.abstract Transport of membrane-­bound organelles is critical to neuronal cell function yet mechanisms hitching vesicles to transport machinery remain elusive. Here we test whether jun-­‐kinase interacting protein (JIP-­‐1), a peripheral membrane scaffolding protein that binds kinesin light chain, is sufficient to mediate cargo transport in axons and study its competition with amyloid precursor protein cytoplasmic domain (APP-­‐C) and negative charge, also known motor receptors. Fluorescent beads (100 nm diameter) exhibit sequence-­‐specific fast anterograde transport (0.46μm/s instantaneous velocity) in the squid giant axon when conjugated to a 14-­‐ amino acid synthetic peptide derived from the carboxyl terminus of JIP-­‐1. JIP-­‐1-­‐ beads have statistically significant faster velocities, longer run lengths, and fewer pauses of shorter durations than APP-­‐C or negatively charged beads by cumulative probability analyses of thousands of motile beads compared by a nonparametric K-­‐S test, with a P=0.004. In competition experiments negatively charged beads gradually cease moving when co-­‐injected with either APP-­‐C or JIP-­‐1 beads, which sustain 90% motility. Co-­‐injection of APP-­‐C and JIP-­‐1 beads decreases each bead's propensity to move initially. At later time points JIP-­‐1-­‐beads recover frequency without further decreasing APP-­‐C moves, suggesting JIP-­‐1 recruits motors from a cryptic pool not accessible to APP-­‐C. Soluble JIP-­‐1 peptide inhibits JIP-­‐1 beads, with smaller effects on APP-­‐C and negatively charged beads. Thus the hierarchy for recruitment of transport machinery is JIP>APP>negative charge. Organelle transport may in part be regulated through the numbers, types and affinities of motor receptors displayed on each organelle's cytoplasmic surface. en_US
dc.description.sponsorship INCBN IGERT and the NIH en_US
dc.language.iso en_US en_US
dc.subject Axonal Transport, JIP-1, APP-C en_US
dc.subject.lcsh Axonal transport.
dc.subject.lcsh Neurons--Physiology.
dc.subject.lcsh Squids--Nervous system.
dc.subject.lcsh Squids--Physiology.
dc.subject.lcsh Kinesin.
dc.title Spatiotemporal dynamics of transport in the squid giant axon : competition between cargo motor receptors, JIP-1, APP-C, and negative charge en_US
dc.type Thesis en_US
dc.description.degree Biomedical Sciences en_US
dc.description.level Masters en_US
dc.description.department University of New Mexico. Biomedical Sciences Graduate Program en_US
dc.description.advisor Bearer, Elaine
dc.description.committee-member Bearer, Elaine
dc.description.committee-member Koch, Steve
dc.description.committee-member Lidke, Diane
dc.description.committee-member Shuttleworth, Bill
dc.description.committee-member Steinberg, Stan
emb.embargo.terms 2013-12-31


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