Use of scanning ion conductance microscopy to guide and redirect neuronal growth cones.
Scanning ion conductance microscopy has been applied to neuronal growth cones of the leech either to image or to stimulate them. Growth cone advance was recorded in non-contact mode using a 2% ion current decrease criterion for pipette-membrane distance control. We demonstrate effective growth cone remodelling using a 5% criterion (near-scanning). Recurrent line near-scanning aligned growth cone processes along the scan line. The new membrane protrusions, marked by DiI, started a few minutes after scanning onset and progressively grew in thickness. Using scanning patterns suitable for connecting distinct growth cones, new links were consistently developed. Although the underlying mechanism is still a matter for investigation, a mechanical perturbation produced by the moving probe appeared to induce the process formation. Thanks to its deterministic and interactive features, this novel approach to guiding growth cones is a promising way to develop networks of identified neurons as well as link them with artificial structures.
Pellegrino
Mario
M
Dipartimento di Scienze Fisiologiche, Università di Pisa, Pisa, Italy. marpell@dfb.unipi.it
Orsini
Paolo
P
De Gregorio
Francesca
F
D000818:Animals; D002465:Cell Movement; D022081:Cell Surface Extensions / Q000502:physiology / Q000648:ultrastructure; D020439:Growth Cones / Q000502:physiology* / Q000648:ultrastructure; D048091:Guided Tissue Regeneration / Q000379:methods; D047709:Hirudo medicinalis / Q000502:physiology / Q000648:ultrastructure; D020527:Microscopy, Scanning Probe / Q000379:methods*; D009416:Nerve Regeneration