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PLoS One. 2012;7(1):e29869. doi: 10.1371/journal.pone.0029869. Epub 2012 Jan 23.

Microfluidic chips for in vivo imaging of cellular responses to neural injury in Drosophila larvae.

Author information

1
Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, United States of America.

Abstract

With powerful genetics and a translucent cuticle, the Drosophila larva is an ideal model system for live imaging studies of neuronal cell biology and function. Here, we present an easy-to-use approach for high resolution live imaging in Drosophila using microfluidic chips. Two different designs allow for non-invasive and chemical-free immobilization of 3(rd) instar larvae over short (up to 1 hour) and long (up to 10 hours) time periods. We utilized these 'larva chips' to characterize several sub-cellular responses to axotomy which occur over a range of time scales in intact, unanaesthetized animals. These include waves of calcium which are induced within seconds of axotomy, and the intracellular transport of vesicles whose rate and flux within axons changes dramatically within 3 hours of axotomy. Axonal transport halts throughout the entire distal stump, but increases in the proximal stump. These responses precede the degeneration of the distal stump and regenerative sprouting of the proximal stump, which is initiated after a 7 hour period of dormancy and is associated with a dramatic increase in F-actin dynamics. In addition to allowing for the study of axonal regeneration in vivo, the larva chips can be utilized for a wide variety of in vivo imaging applications in Drosophila.

PMID:
22291895
PMCID:
PMC3264548
DOI:
10.1371/journal.pone.0029869
[Indexed for MEDLINE]
Free PMC Article

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