Ultra-short pulses to signal neuronal growth cone machinery

Measurable change in the sensory motor machinery of growth cones are induced by non contact femtosecond laser. The focused laser beam with an average power of 3 mW was positioned at some distance away from the closest fillopodia of cortical neurons from primary cell cultures (mice E15). By identifying a set of preliminary parameters we were able to statistically analyze the phenomenological behavior of the fillopodia and classify the effects different conditions of laser light has on the growth cone. Results show that fillopodia become significantly biased towards the focused femtosecond laser light. The same experiment performed with continuous wave (CW) produced results which were indistinguishable from the case where there is no laser light present (placebo condition) indicating no clear effects of the CW laser light on the fillopodia at a distance. These findings show the potential for ultrashort pulsed light to become a new type of pathfinding cue for neuronal growth cones.

[1]  Astrid A. Prinz,et al.  Electrical synapses by guided growth of cultured neurons from the snail Lymnaea stagnalis , 2000, Biological Cybernetics.

[2]  J. Goldberg.  How does an axon grow? , 2003, Genes & development.

[3]  Marie-France Carlier,et al.  Mechanism of Actin-Based Motility , 2001, Science.

[4]  Makiko Sato,et al.  Guiding neuronal development with in situ microfabrication , 2004 .

[5]  T. Svitkina,et al.  Actin machinery: pushing the envelope. , 2000, Current opinion in cell biology.

[6]  P. Forscher,et al.  Substrate-cytoskeletal coupling as a mechanism for the regulation of growth cone motility and guidance. , 2000, Journal of neurobiology.

[7]  Rafael Yuste,et al.  Multiphoton stimulation of neurons. , 2002, Journal of neurobiology.

[8]  A. B. Huber,et al.  Signaling at the growth cone: ligand-receptor complexes and the control of axon growth and guidance. , 2003, Annual review of neuroscience.

[9]  J. Käs,et al.  Guiding neuronal growth with light , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Samarendra K Mohanty,et al.  Controlled induction, enhancement, and guidance of neuronal growth cones by use of line optical tweezers. , 2005, Optics letters.

[11]  D. Jay,et al.  The clutch hypothesis revisited: ascribing the roles of actin-associated proteins in filopodial protrusion in the nerve growth cone. , 2000, Journal of neurobiology.

[12]  Peter Fromherz,et al.  Guided outgrowth of leech neurons in culture , 1991, Neuroscience Letters.

[13]  J. Chilton.  Molecular mechanisms of axon guidance. , 2006, Developmental biology.

[14]  J. Kornhuber,et al.  Light‐induced exocytosis in cell development and differentiation , 2006, Journal of cellular biochemistry.

[15]  M. Poo,et al.  Orientation of neurite growth by extracellular electric fields , 1982, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[16]  Ignacio Tinoco,et al.  Temperature control methods in a laser tweezers system. , 2005, Biophysical journal.

[17]  M. Kirschner,et al.  Cytoskeletal dynamics and nerve growth , 1988, Neuron.

[18]  C. Goodman,et al.  The Molecular Biology of Axon Guidance , 1996, Science.

[19]  G. Albrecht‐Buehler,et al.  Surface extensions of 3T3 cells towards distant infrared light sources , 1991, The Journal of cell biology.