Research Papers

Rat embryonic hippocampus and induced pluripotent stem cell derived cultured neurons recover from laser-induced subaxotomy

[+] Author Affiliations
Aaron Selfridge, Nicholas Hyun

University of California, San Diego, Department of Bioengineering, 9500 Gilman Drive, La Jolla, California 92093, United States

Chai-Chun Chiang, April M. Weissmiller, William C. Mobley

University of California, San Diego, Department of Neurosciences, 9500 Gilman Drive, La Jolla, California 92093, United States

Sol M. Reyna

University of California, San Diego, Department of Biomedical Sciences, 9500 Gilman Drive, La Jolla, California 92093, United States

Linda Z. Shi

University of California, San Diego, Institute of Engineering in Medicine, 9500 Gilman Drive, La Jolla, California 92093, United States

Daryl Preece

University of California, San Diego, Department of NanoEngineering, 9500 Gilman Drive La Jolla, California 92093, United States

Michael W. Berns

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

Neurophoton. 2(1), 015006 (Feb 13, 2015). doi:10.1117/1.NPh.2.1.015006
History: Received September 28, 2014; Accepted January 12, 2015
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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 Nd:YVO4 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.

Figures in this Article
© 2015 Society of Photo-Optical Instrumentation Engineers

Topics

Lasers ; Nerve ; Axon ; Stem cells

Citation

Aaron Selfridge ; Nicholas Hyun ; Chai-Chun Chiang ; Sol M. Reyna ; April M. Weissmiller, et al.
"Rat embryonic hippocampus and induced pluripotent stem cell derived cultured neurons recover from laser-induced subaxotomy", Neurophoton. 2(1), 015006 (Feb 13, 2015). ; http://dx.doi.org/10.1117/1.NPh.2.1.015006


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