Use of optical tweezers technology for long-term, focal stimulation of specific subcellular neuronal compartments

D'Este, Elisa, Baj, Gabriele, Beuzer, Paolo , Ferrari, Enrico, Pinato, Giulietta, Tongiorgi, Enrico and Cojoc, Dan (2011) Use of optical tweezers technology for long-term, focal stimulation of specific subcellular neuronal compartments. Integrative Biology, 3 (5). pp. 568-577. ISSN 1757-9694

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Spatial regulation of secretory molecule release is a sophisticated mechanism used by the nervous system to control network development and finely tune the activity of each synapse. Great efforts have been made to develop techniques that mimic secretory molecule release with the aim of stimulating neurons as close as possible to physiological conditions. However, current techniques have poor spatial resolution or low flexibility. Here, we propose a novel approach to achieve focal and prolonged stimulation of neurons using optical tweezers and single microbeads functionalized with a secretory molecule, the neurotrophin brain-derived neurotrophic factor (BDNF). BDNF is a key regulator of neuronal development and plasticity. We show that single BDNF-coated microbeads can be extracted with optical tweezers from small reservoirs and positioned with submicrometric precision to specific sites on the dendrites of cultured hippocampal neurons. Localized contact of microbeads functionalized with BDNF, but not with bovine serum albumin (BSA), induced focal increase of calcium signaling in the stimulated dendrite, specific activation of the TrkB receptor pathway and influenced the development of growth cones. Remarkably, a single BDNF-coated bead localized on a dendrite was found to be enough for TrkB phosphorylation, an efficient and long-lasting activation of calcium signaling in the soma, and c-Fos signaling in the nucleus, comparable to bath stimulation conditions. These findings support the use of optical tweezer technology for long-term, localized stimulation of specific subcellular neuronal compartments.

Keywords:Optical tweezers, brain-derived neurotrophic factor
Subjects:B Subjects allied to Medicine > B140 Neuroscience
C Biological Sciences > C700 Molecular Biology, Biophysics and Biochemistry
Divisions:College of Science > School of Life Sciences
ID Code:6581
Deposited On:15 Oct 2012 20:51

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