title: Rat embryonic hippocampus and induced pluripotent stem cell derived cultured neurons recover from laser-induced subaxotomy.
abstract: Axonal injury and stress have long been thought to play a pathogenic role in a variety of neurodegenerative diseases. However, a model for studying single-cell axonal injury in mammalian cells and the processes of repair has not been established. The purpose of this study was to examine the response of neuronal growth cones to laser-induced axonal damage in cultures of embryonic rat hippocampal neurons and induced pluripotent stem cell (iPSC) derived human neurons. A 532-nm pulsed [Formula: see text] picosecond laser was focused to a diffraction limited spot at a precise location on an axon using a laser energy/power that did not rupture the cell membrane (subaxotomy). Subsequent time series images were taken to follow axonal recovery and growth cone dynamics. After laser subaxotomy, axons thinned at the damage site and initiated a dynamic cytoskeletal remodeling process to restore axonal thickness. The growth cone was observed to play a role in the repair process in both hippocampal and iPSC-derived neurons. Immunofluorescence staining confirmed structural tubulin damage and revealed initial phases of actin-based cytoskeletal remodeling at the damage site. The results of this study indicate that there is a repeatable and cross-species repair response of axons and growth cones after laser-induced damage.
authors.affiliation: University of California , San Diego, Department of Bioengineering, 9500 Gilman Drive, La Jolla, California 92093, United States.
authors.forename: Aaron
authors.initials: A
authors.lastname: Selfridge
authors.affiliation: University of California , San Diego, Department of Bioengineering, 9500 Gilman Drive, La Jolla, California 92093, United States.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Hyun
authors.affiliation: University of California , San Diego, Department of Neurosciences, 9500 Gilman Drive, La Jolla, California 92093, United States.
authors.forename: Chai-Chun
authors.initials: CC
authors.lastname: Chiang
authors.affiliation: University of California , San Diego, Department of Biomedical Sciences, 9500 Gilman Drive, La Jolla, California 92093, United States.
authors.forename: Sol M
authors.initials: SM
authors.lastname: Reyna
authors.affiliation: University of California , San Diego, Department of Neurosciences, 9500 Gilman Drive, La Jolla, California 92093, United States.
authors.forename: April M
authors.initials: AM
authors.lastname: Weissmiller
authors.affiliation: University of California , San Diego, Institute of Engineering in Medicine, 9500 Gilman Drive, La Jolla, California 92093, United States.
authors.forename: Linda Z
authors.initials: LZ
authors.lastname: Shi
authors.affiliation: University of California , San Diego, Department of NanoEngineering, 9500 Gilman Drive La Jolla, California 92093, United States.
authors.forename: Daryl
authors.initials: D
authors.lastname: Preece
authors.affiliation: University of California , San Diego, Department of Neurosciences, 9500 Gilman Drive, La Jolla, California 92093, United States.
authors.forename: William C
authors.initials: WC
authors.lastname: Mobley
authors.affiliation: University of California , San Diego, Department of Bioengineering, 9500 Gilman Drive, La Jolla, California 92093, United States ; University of California , San Diego, Institute of Engineering in Medicine, 9500 Gilman Drive, La Jolla, California 92093, United States ; University of California , Irvine, Beckman Laser Institute, 1002 Health Sciences Road, Irvine, California 92612, United States.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Berns
keywords: growth cone; hippocampus; induced pluripotent stem cell; neurons; regeneration; subaxotomy