To understand how the microtubule motors, cytoplasmic dynein and kinesins, find and transport cargoes to specific cellular destinations, we need to know much more about how motors are attached to cargoes. The extended morphology of axons and dendrites makes post-mitotic neurons especially dependent on polarized transport and ideal for studying the regulation of transport. We recently showed that ss-spectrin mutations in humans that cause spinocerebellar ataxia type 5 (SCA5) impair motor-based transport of synaptic vesicles in a Drosophila model of neurodegeneration. In this application, we focus on an analysis of ss-spectrin and its role in mediating the attachment of dynactin and cytoplasmic dynein to cargoes in neurons. We will characterize the defective neuronal transport in our fly model and will pursue the gene products and mechanisms that regulate transport. The selective use of the proposed spectrin/ motor linkage in will be studied in the axons and dendrites of living neurons (Aim 1). New genetic modifiers of the SCA5 phenotypes and neuronal transport will be characterized in motility and biochemical assays (Aim 2), and the connection between neuronal transport, autophagy and neurodegeneration will be explored (Aim 3). Our work has important long-term implications for the understanding and treatment of spinal cord injuries, and the family of neurodegenerative diseases, including Amyotrophic lateral sclerosis, Huntington's Disease, and Alzheimer's Disease.

Public Health Relevance

We are studying the molecular basis of intracellular transport. Our work focuses on the microtubule motors cytoplasmic dynein and kinesin and their function in neurons. We have developed a fly model for neurodegeneration in the disease Spinocerebellar ataxia type 5 and we will study how perturbations in intracellular transport contribute to neurodegenerative disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM044757-22
Application #
8549260
Study Section
Nuclear and Cytoplasmic Structure/Function and Dynamics Study Section (NCSD)
Program Officer
Gindhart, Joseph G
Project Start
1990-07-01
Project End
2015-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
22
Fiscal Year
2013
Total Cost
$322,243
Indirect Cost
$108,837
Name
University of Minnesota Twin Cities
Department
Genetics
Type
Schools of Medicine
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Neisch, Amanda L; Avery, Adam W; Machamer, James B et al. (2016) Methods to identify and analyze gene products involved in neuronal intracellular transport using Drosophila. Methods Cell Biol 131:277-309
Mauvezin, Caroline; Neisch, Amanda L; Ayala, Carlos I et al. (2016) Coordination of autophagosome-lysosome fusion and transport by a Klp98A-Rab14 complex in Drosophila. J Cell Sci 129:971-82
Avery, Adam W; Crain, Jonathan; Thomas, David D et al. (2016) A human β-III-spectrin spinocerebellar ataxia type 5 mutation causes high-affinity F-actin binding. Sci Rep 6:21375
Delaney, Susan K; Hultner, Michael L; Jacob, Howard J et al. (2016) Toward clinical genomics in everyday medicine: perspectives and recommendations. Expert Rev Mol Diagn 16:521-32
Materassi, Donatello; Roychowdhury, Subhrajit; Hays, Thomas et al. (2013) An exact approach for studying cargo transport by an ensemble of molecular motors. BMC Biophys 6:14
Aggarwal, Tanuj; Materassi, Donatello; Davison, Robert et al. (2012) Detection of Steps in Single Molecule Data. Cell Mol Bioeng 5:14-31
Fasulo, Barbara; Koyama, Carol; Yu, Kristina R et al. (2012) Chk1 and Wee1 kinases coordinate DNA replication, chromosome condensation, and anaphase entry. Mol Biol Cell 23:1047-57
Smith, Rebecca B; Machamer, James B; Kim, Nam Chul et al. (2012) Relay of retrograde synaptogenic signals through axonal transport of BMP receptors. J Cell Sci 125:3752-64
Reis, Gerald F; Yang, Ge; Szpankowski, Lukasz et al. (2012) Molecular motor function in axonal transport in vivo probed by genetic and computational analysis in Drosophila. Mol Biol Cell 23:1700-14
Tao, Xiaodong; Crest, Justin; Kotadia, Shaila et al. (2012) Live imaging using adaptive optics with fluorescent protein guide-stars. Opt Express 20:15969-82

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