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Proceedings Paper

In vivo imaging of neural reactive plasticity after laser axotomy in cerebellar cortex
Author(s): A. L. Allegra Mascaro; L. Sacconi; B. Maco; G. W. Knott; F. S. Pavone
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Paper Abstract

Multi-photon imaging provides valuable insights into the continuous reshaping of neuronal connectivity in live brain. We previously showed that single neuron or even single spine ablation can be achieved by laser-mediated dissection. Furthermore, single axonal branches can be dissected avoiding collateral damage to the adjacent dendrite and the formation of a persistent glial scar. Here, we describe the procedure to address the structural plasticity of cerebellar climbing fibers by combining two-photon in vivo imaging with laser axotomy in a mouse model. This method is a powerful tool to study the basic mechanisms of axonal rewiring after single branch axotomy in vivo. In fact, despite the denervated area being very small, the injured axons consistently reshape the connectivity with surrounding neurons, as indicated by the increase in the turnover of synaptic boutons. In addition, time-lapse imaging reveals the sprouting of new branches from the injured axon. Newly formed branches with varicosities suggest the possible formation of synaptic contacts. Correlative light and electron microscopy revealed that the sprouted branch contains large numbers of vesicles, with varicosities in the close vicinity of Purkinje dendrites.

Paper Details

Date Published: 24 March 2014
PDF: 6 pages
Proc. SPIE 8928, Optical Techniques in Neurosurgery, Neurophotonics, and Optogenetics, 89281N (24 March 2014); doi: 10.1117/12.2037276
Show Author Affiliations
A. L. Allegra Mascaro, European Lab. for Non-Linear Spectroscopy, Univ. of Florence (Italy)
L. Sacconi, European Lab. for Non-Linear Spectroscopy, Univ. of Florence (Italy)
INO-CNR (Italy)
B. Maco, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
G. W. Knott, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
F. S. Pavone, European Lab. for Non-Linear Spectroscopy, Univ. of Florence (Italy)
INO-CNR (Italy)
ICON Foundation (Italy)


Published in SPIE Proceedings Vol. 8928:
Optical Techniques in Neurosurgery, Neurophotonics, and Optogenetics
Henry Hirschberg; E. Duco Jansen; Samarendra K. Mohanty; Nitish V. Thakor; Qingming Luo; Steen J. Madsen, Editor(s)

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