Nano-volume drop patterning for rapid on-chip neuronal connect-ability assays.
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| Abstract |
The ability of neurons to extend projections and to form physical connections among them (i.e., "connect-ability") is altered in several neuropathologies. The quantification of these alterations is an important read-out to investigate pathogenic mechanisms and for research and development of neuropharmacological therapies, however current morphological analysis methods are very time-intensive. Here, we present and characterize a novel on-chip approach that we propose as a rapid assay. Our approach is based on the definition on a neuronal cell culture substrate of discrete patterns of adhesion protein spots (poly-d-lysine, 23 ± 5 μm in diameter) characterized by controlled inter-spot separations of increasing distance (from 40 μm to 100 μm), locally adsorbed in an adhesion-repulsive agarose layer. Under these conditions, the connect-ability of wild type primary neurons from rodents is shown to be strictly dependent on the inter-spot distance, and can be rapidly documented by simple optical read-outs. Moreover, we applied our approach to identify connect-ability defects in neurons from a mouse model of 22q11.2 deletion syndrome/DiGeorge syndrome, by comparative trials with wild type preparations. The presented results demonstrate the sensitivity and reliability of this novel on-chip-based connect-ability approach and validate the use of this method for the rapid assessment of neuronal connect-ability defects in neuropathologies.
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| Authors | Alessia Petrelli, Emanuele Marconi, Marco Salerno, Davide De Pietri Tonelli, Luca Berdondini, Silvia Dante |
| Journal | Lab on a chip (Lab Chip) Vol. 13 Issue 22 Pg. 4419-29 (Nov 21 2013) ISSN: 1473-0189 [Electronic] England |
| PMID | 24064674 (Publication Type: Journal Article) |
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