Design and Implementation of in Vivo Imaging of Neural Injury Responses in the Adult Drosophila Wing

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2013 Apr 18

PMID 23589940

Citations 20

Authors

Yanshan Fang

Lorena Soares

Nancy M Bonini

Affiliations

Soon will be listed here.

Abstract

Live-imaging technology has markedly advanced in the field of neural injury and axon degeneration; however, studies are still predominantly performed in in vitro settings such as cultured neuronal cells or in model organisms such as Caenorhabditis elegans in which axons lack glial wrappings. We recently developed a new in vivo model for adult-stage neural injury in Drosophila melanogaster, using the highly accessible wing of the animal. Because the Drosophila wing is translucent and dispensable for survival, it allows clear and direct visualization of injury-induced progressive responses of axons and glia highlighted by fluorescent protein (FP) markers in live animals over time. Moreover, unlike previous Drosophila models of neural injury, this procedure does not require dissection of the CNS. Thus, the key preparation steps for in vivo imaging of the neural injury response described in this protocol can be completed within 30 min.

Citing Articles

Study of Axonal Injury and Degeneration in .

Waller T, Smithson L, Collins C Cold Spring Harb Protoc. 2024; .

PMID: 38649194 PMC: 11701930. DOI: 10.1101/pdb.top108163.

Wallerian Degeneration and Clearance of Olfactory Receptor Neuron Axons following Antennal Transection.

Waller T, Smithson L, Collins C Cold Spring Harb Protoc. 2024; .

PMID: 38649191 PMC: 11701940. DOI: 10.1101/pdb.prot108167.

Detailed Imaging of Mitochondrial Transport and Precise Manipulation of Mitochondrial Function with Genetically Encoded Photosensitizers in Adult Drosophila Neurons.

Mattedi F, Chennell G, Vagnoni A Methods Mol Biol. 2022; 2431:385-407.

PMID: 35412288 DOI: 10.1007/978-1-0716-1990-2_20.

Current methods to analyze lysosome morphology, positioning, motility and function.

Barral D, Staiano L, Guimas Almeida C, Cutler D, Eden E, Futter C Traffic. 2022; 23(5):238-269.

PMID: 35343629 PMC: 9323414. DOI: 10.1111/tra.12839.

Establishment of a AD model.

Wang X, Zhao Y, Hu Y, Ren P, Sun Y, Guo X J Biol Methods. 2019; 3(2):e43.

PMID: 31453210 PMC: 6706136. DOI: 10.14440/jbm.2016.61.

References

Wang J, Beck E, McCabe B . A modular toolset for recombination transgenesis and neurogenetic analysis of Drosophila. PLoS One. 2012; 7(7):e42102. PMC: 3405054. DOI: 10.1371/journal.pone.0042102. View

El Bejjani R, Hammarlund M . Neural regeneration in Caenorhabditis elegans. Annu Rev Genet. 2012; 46:499-513. PMC: 3700416. DOI: 10.1146/annurev-genet-110711-155550. View

Fang Y, Soares L, Teng X, Geary M, Bonini N . A novel Drosophila model of nerve injury reveals an essential role of Nmnat in maintaining axonal integrity. Curr Biol. 2012; 22(7):590-5. PMC: 3347919. DOI: 10.1016/j.cub.2012.01.065. View

Becker C, Becker T . Adult zebrafish as a model for successful central nervous system regeneration. Restor Neurol Neurosci. 2008; 26(2-3):71-80. View

Rosenfeld J, Maas A, Bragge P, Morganti-Kossmann M, Manley G, Gruen R . Early management of severe traumatic brain injury. Lancet. 2012; 380(9847):1088-98. DOI: 10.1016/S0140-6736(12)60864-2. View

Hoopfer E, McLaughlin T, Watts R, Schuldiner O, OLeary D, Luo L . Wlds protection distinguishes axon degeneration following injury from naturally occurring developmental pruning. Neuron. 2006; 50(6):883-95. DOI: 10.1016/j.neuron.2006.05.013. View

Potter C, Turenchalk G, Xu T . Drosophila in cancer research. An expanding role. Trends Genet. 2000; 16(1):33-9. DOI: 10.1016/s0168-9525(99)01878-8. View

. Postembryonic sensory axon guidance in Drosophila. Cell Mol Life Sci. 1999; 55(11):1386-98. PMC: 11146940. DOI: 10.1007/s000180050379. View

Lessing D, Bonini N . Maintaining the brain: insight into human neurodegeneration from Drosophila melanogaster mutants. Nat Rev Genet. 2009; 10(6):359-70. PMC: 2820605. DOI: 10.1038/nrg2563. View

10.

Ghosh-Roy A, Chisholm A . Caenorhabditis elegans: a new model organism for studies of axon regeneration. Dev Dyn. 2010; 239(5):1460-4. PMC: 3069807. DOI: 10.1002/dvdy.22253. View

11.

Toomre D, Bewersdorf J . A new wave of cellular imaging. Annu Rev Cell Dev Biol. 2010; 26:285-314. DOI: 10.1146/annurev-cellbio-100109-104048. View

12.

Cho S, Wood A, Bowlby M . Brain slices as models for neurodegenerative disease and screening platforms to identify novel therapeutics. Curr Neuropharmacol. 2008; 5(1):19-33. PMC: 2435340. DOI: 10.2174/157015907780077105. View

13.

Sehgal A, Joiner W, Crocker A, Koh K, Sathyanarayanan S, Fang Y . Molecular analysis of sleep: wake cycles in Drosophila. Cold Spring Harb Symp Quant Biol. 2008; 72:557-64. DOI: 10.1101/sqb.2007.72.018. View

14.

Taylor A, Blurton-Jones M, Rhee S, Cribbs D, Cotman C, Jeon N . A microfluidic culture platform for CNS axonal injury, regeneration and transport. Nat Methods. 2005; 2(8):599-605. PMC: 1558906. DOI: 10.1038/nmeth777. View

15.

Keene A, Waddell S . Drosophila olfactory memory: single genes to complex neural circuits. Nat Rev Neurosci. 2007; 8(5):341-54. DOI: 10.1038/nrn2098. View

16.

Brand A, Perrimon N . Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development. 1993; 118(2):401-15. DOI: 10.1242/dev.118.2.401. View

17.

Xiong X, Wang X, Ewanek R, Bhat P, DiAntonio A, Collins C . Protein turnover of the Wallenda/DLK kinase regulates a retrograde response to axonal injury. J Cell Biol. 2010; 191(1):211-23. PMC: 2953441. DOI: 10.1083/jcb.201006039. View

18.

MacDonald J, Beach M, Porpiglia E, Sheehan A, Watts R, Freeman M . The Drosophila cell corpse engulfment receptor Draper mediates glial clearance of severed axons. Neuron. 2006; 50(6):869-81. DOI: 10.1016/j.neuron.2006.04.028. View

19.

Lai S, Lee T . Genetic mosaic with dual binary transcriptional systems in Drosophila. Nat Neurosci. 2006; 9(5):703-9. DOI: 10.1038/nn1681. View

20.

Pfeiffer B, Ngo T, Hibbard K, Murphy C, Jenett A, Truman J . Refinement of tools for targeted gene expression in Drosophila. Genetics. 2010; 186(2):735-55. PMC: 2942869. DOI: 10.1534/genetics.110.119917. View