Microfluidic Chips for In Vivo Imaging of Cellular Responses to Neural Injury in Drosophila Larvae
نویسندگان
چکیده
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.
منابع مشابه
Concentration dependent effect of morphine, aspirin, capsaicin and chili pepper hydro alcoholic extract on thermal and chemical pain model in fruit fly (Drosophila melanogaster)
Introduction: Pain research using animal models is related to ethical concerns, so invertebrates and insects have been recommended by researchers. In the present study, the nociceptive and antinociceptive effects of capsaicin, aspirin, morphine and chili extract were examined using fruit fly (Drosophila melanogaster) as an alternative for rodent pain model. Methods: Stage 3 of larvae and ad...
متن کاملUsing Microfluidics Chips for Live Imaging and Study of Injury Responses in Drosophila Larvae
Live imaging is an important technique for studying cell biological processes, however this can be challenging in live animals. The translucent cuticle of the Drosophila larva makes it an attractive model organism for live imaging studies. However, an important challenge for live imaging techniques is to noninvasively immobilize and position an animal on the microscope. This protocol presents a...
متن کاملMicrofluidic Devices for Anesthetic Free In-vivo Axonal Transport Imaging
Microfluidic devices have been used for in-vitro studies in the past while recently they have been extended for in-vivo studies in whole organisms. In this report we use a microfluidic device for high resolution fluorescence imaging of axonal transport in model systems Caenorhabditis elegans and Drosophila larvae. The device immobilized animals are compared with respect to the anesthetized anim...
متن کاملAdministration of Leukemia Inhibitory Factor Increases Opalin Expression in the Cerebral Cortex of Male Balb/C Mice An In Vivo Study
Background: Leukemia inhibitory factor (LIF) is a neurortophic cytokine which plays an important role in the neural cell survival. Expression of LIF and its receptor, LIFR, in different brain regions has been demonstrated. Based on evidences LIF plays an important role in the modulation of neurogenesis and glial responses to injury. Up-regulation of LIF after central nervous system (CNS) damage...
متن کاملToxicological Evaluation of a New Lepidopteran Insecticide, Flubendiamide, in Non-Target Drosophila melanogaster Meigen (Diptera: Drosophilidae)
Background: Flubendiamide, comparatively a new pesticide designed to eradicate lepidopteran insect pests is known to have low risk to birds, mammals, fish, algae, honey bees, non-target arthropods, earthworms, soil macro- and micro-organisms, non-target plants as well as sewage treatment organisms; however, the risk assessment for aquatic invertebrates from metabolite could not be finalized wit...
متن کامل