Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by the selective loss of motor neurons from the brain and spinal cord. Disease progression is rapid and patients typically die from respiratory failure 3-5 years following disease onset. Numerous ALS causing mutant genes have been identified but the underlying cause of motor neuron degeneration is not known. Recent genome analysis from families that contain a heritable form of ALS have identified multiple mutations in two cytoskeletal genes, profilin 1 and TUBA4A, that cause ALS; linking cytoskeletal dysfunction to ALS pathology. In this proposal I will examine whether an ALS mutant gene disrupts two cytoskeletal-dependent functions that are crucial for motor neuron health. Motor neurons extend structures great distances from the cell body and rely heavily on the microtubule cytoskeleton for the transport of cellular materials to these distal structures. In the first part of the proposl, I will examine whether expression of mutant TUBA4A affects the transport of essential microtubule based cargos to and from the distal structures. I first will use live cell imaging to track the rate of transport of various essential cargos in primary cultured neurons expressing mutant TUBA4A. Next I will examine if developmental or cellular interactions play a contributing role by examining changes in motor protein movement in the sciatic nerve of mutant TUBA4A expressing mice. Finally, I will evaluate weather mutant TUBA4A expression changes the motor neuron tolerance to microtubule-destabilizing neurotoxic insult. Injury to motor neurons causes separation of the motor neuron axon from the neuromuscular junction (NMJ). After the initial denervation of the NMJ the motor neuron can follow one of two paths: axon regrowth and reattachment to the NMJ or continued axon dieback and cell death. The stability of the microtubules in the tip of the injured axon will influence which of these two paths will be the finl outcome. In the second part of the proposal I will examine if mutant TUBA4A expression increases retraction bulb formation and decreases axon regeneration following injury. I will first use live cell imaging to measure the rate of regrowth of severed axons in primary motor neurons expressing mutant TUBA4A. I will next examine whether mutant TUBA4A expression changes the rate of growth cone formation and microtubule organization after sciatic nerve lesion in mutant TUBA4A expressing mice. Finally, I will examine whether mutant TUBA4A affects the reinnervation of muscle tissue following nerve crush in mutant TUBA4A expressing mice. The goal of this proposal is to further knowledge on how cytoskeleton-dependent functions are involved in ALS disease pathology. The results gained from this proposal will offer new insight into the cause of cell death in ALS and will open new areas for drug research and therapy.

Public Health Relevance

Amyotrophic lateral sclerosis (ALS) is a fatal neurological disease that affects 20,000-30,000 of the U.S. population. The disease attacks the nerve cells (motor neurons) that control voluntary muscle action. ALS causes a loss a loss of muscle coordination, progressive muscle weakness and eventually paralysis and death. Disease progression is rapid and patients typically die within 3-4 years from respiratory failure following the initial diagnosis. Numerous mutant genes have been found to cause ALS, however, the mechanism leading to disease onset is not fully understood. Recently multiple mutations in two cytoskeletal genes profilin 1 and TUBA4A implicate cytoskeletal defects as contributing factors to ALS. Additionally, mouse models of ALS show defective cytoskeletal-based intracellular transport of materials within motor neurons emphasizing the importance the cytoskeleton. The proposed research aims to examine in more detail how disruption of the cytoskeleton contributes to ALS disease pathology.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32NS090831-03
Application #
9221364
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Gubitz, Amelie
Project Start
2015-04-01
Project End
2018-03-31
Budget Start
2017-04-01
Budget End
2018-03-31
Support Year
3
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Massachusetts Medical School Worcester
Department
Neurology
Type
Schools of Medicine
DUNS #
603847393
City
Worcester
State
MA
Country
United States
Zip Code
01655