title: Measurement of subcellular force generation in neurons.
abstract: Forces are important for neuronal outgrowth during the initial wiring of the nervous system and after trauma, yet subcellular force generation over the microtubule-rich region at the rear of the growth cone and along the axon has never, to our knowledge, been directly measured. Because previous studies have indicated microtubule polymerization and the microtubule-associated proteins Kinesin-1 and dynein all generate forces that push microtubules forward, a major question is whether the net forces in these regions are contractile or expansive. A challenge in addressing this is that measuring local subcellular force generation is difficult. Here we develop an analytical mathematical model that describes the relationship between unequal subcellular forces arranged in series within the neuron and the net overall tension measured externally. Using force-calibrated towing needles to measure and apply forces, in combination with docked mitochondria to monitor subcellular strain, we then directly measure force generation over the rear of the growth cone and along the axon of chick sensory neurons. We find the rear of the growth cone generates 2.0 nN of contractile force, the axon generates 0.6 nN of contractile force, and that the net overall tension generated by the neuron is 1.3 nN. This work suggests that the forward bulk flow of the cytoskeletal framework that occurs during axonal elongation and growth-cone pauses arises because strong contractile forces in the rear of the growth cone pull material forward.
authors.affiliation: Department of Mathematics, Kettering University, Flint, Michigan.
authors.forename: Matthew
authors.initials: M
authors.lastname: O'Toole
authors.affiliation: Department of Zoology, Michigan State University, East Lansing, Michigan.
authors.forename: Phillip
authors.initials: P
authors.lastname: Lamoureux
authors.affiliation: Department of Zoology, Michigan State University, East Lansing, Michigan. Electronic address: kmiller@msu.edu.
authors.forename: Kyle E
authors.initials: KE
authors.lastname: Miller
mesh_terms: D000818:Animals; D001369:Axons / Q000378:metabolism; D002478:Cells, Cultured; D002642:Chick Embryo; D003599:Cytoskeleton / Q000378:metabolism; D008954:Models, Biological; D009038:Motion; D011984:Sensory Receptor Cells / Q000378:metabolism* / Q000648:ultrastructure; D013314:Stress, Mechanical