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Published January 23, 2007 | Supplemental Material + Published
Journal Article Open

Differences in protein mobility between pioneer versus follower growth cones

Abstract

Navigating growth cones need to integrate, process and respond to guidance signals, requiring dynamic information transfer within and between different compartments. Studies have shown that, faced with different navigation challenges, growth cones display dynamic changes in growth kinetics and morphologies. However, it remains unknown whether these are paralleled by differences in their internal molecular dynamics. To examine whether there are protein mobility differences during guidance, we developed multiphoton fluorescence recovery after photobleaching methods to determine molecular diffusion rates in pathfinding growth cones in vivo. Actively navigating growth cones (leaders) have consistently longer recovery times than growth cones that are fasciculated and less actively navigating (followers). Pharmacological perturbations of the cytoskeleton point to actin as the primary modulator of diffusion in differently behaving growth cones. This approach provides a powerful means to quantify mobility of specific proteins in neurons in vivo and reveals that diffusion is important during axon navigation.

Additional Information

Copyright © 2007 by the National Academy of Sciences. Communicated by Harry B. Gray, California Institute of Technology, Pasadena, CA, November 15, 2006 (received for review May 10, 2006). Published online before print January 17, 2007, 10.1073/pnas.0610142104 We thank Michael Liebling, Helen McBride, and Marianne Bronner-Fraser for helpful discussion and comments on the manuscript. This work was supported by the Beckman Institute at Caltech and National Institutes of Health Grants HD043897 and HD037105 (to S.E.F.) and a National Defense Science and Engineering Graduate Fellowship (to R.P.K.). Author contributions: R.P.K., M.B.-M., and S.E.F. designed research; R.P.K. and M.B.-M. performed research; R.P.K. contributed new reagents/analytic tools; R.P.K. and M.B.-M. analyzed data; and R.P.K., M.B.-M., and S.E.F. wrote the paper. The authors declare no conflict of interest. This article contains supporting information online at www.pnas.org/cgi/content/full/0610142104/DC1.

Attached Files

Published - 1207.pdf

Supplemental Material - KULpnas07sfig4.pdf

Supplemental Material - KULpnas07sfig5a.pdf

Supplemental Material - KULpnas07sfig5b.pdf

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Supplemental Material - KULpnas07smovie1.avi

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Supplemental Material - KULpnas07smovie3.avi

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August 22, 2023
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