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JoVE Journal Developmental Biology
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Developmental Biology

Published: April 28, 2021 doi: 10.3791/61273
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1, 1, 1, 1
1Departments of Biology and Cell & Systems Biology, University of Toronto - Mississauga

Materials

Name Company Catalog Number Comments
10x PBS Bioshop PBS405
37% formaldehyde Bioshop FOR201
Alexa Fluor 488 (goat) secondary Invitrogen A-11055 use at 1:500
Alexa Fluor 555 (mouse) secondary Invitrogen A-31570 use at 1:500
Alexa Fluor 647 (guinea pig) secondary Invitrogen A-21450 use at 1:500
Alexa Fluor 647 (rat) secondary Invitrogen A-21247 use at 1:500
Cover slips VWR 48366-067
Dissecting forceps - #5 Dumont 11251-10
Dissecting forceps - #55 Dumont 11295-51
Dissection Dish Corning 722085
Dry wipes Kimbery Clark 34155
Goat anti-Bgal primary antibody Biogenesis use at 1:1000
Guinea pig anti-Bsh primary antibody Gift from Claude Desplan use at 1:500
Guinea pig anti-Vsx1 primary antibody Erclik et al. 2008 use at 1:1000
Laboratory film Parafilm PM-996
Microcentrifuge tubes Sarstedt 72.706.600
Microscope slides VWR CA4823-180
Mouse anti-dac primary antibody Developmental Studies Hybridoma Bank (DSHB) mabdac2-3 use at 1:20
Mouse anti-eya primary antibody DSHB eya10H6 use at 1:20
Mouse anti-nc82 primary antibody DSHB nc82 use at 1:50
Mouse anti-svp primary antibody DSHB Seven-up 2D3 use at 1:100
Polymer Clay Any type of clay can be used
Rabbit anti-GFP Invitrogen A-11122 use at 1:1000
Rat anti-DE-Cadherin primary antibody DSHB DCAD2 use at 1:20
Slowfade mounting medium Invitrogen S36967 Vectashield mounting medium ( cat# H-1000) can also be used
Triton-x-100 Bioshop TRX506

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References

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  2. Hofbauer, A., Campos-Ortega, J. A. Proliferation pattern and early differentiation of the optic lobes in Drosophila melanogaster. Roux's Archives of Developmental Biology. 198, 264-274 (1990).
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  4. Nériec, N., Desplan, C. From the Eye to the Brain. Development of the Drosophila Visual System. Current Topics in Developmental Biology. 116, Academic Press Inc. 247-271 (2016).
  5. Apitz, H., Salecker, I. A Challenge of Numbers and Diversity: Neurogenesis in the Drosophila Optic Lobe. Journal of Neurogenetics. 28, 233-249 (2014).
  6. Contreras, E. G., Sierralta, J., Oliva, C. Novel strategies for the generation of neuronal diversity: Lessons from the fly visual system. Frontiers in Molecular Neuroscience. 12, 140 (2019).
  7. Erclik, T., Hartenstein, V., Lipshitz, H. D., McInnes, R. R. Conserved Role of the Vsx Genes Supports a Monophyletic Origin for Bilaterian Visual Systems. Current Biology. 18, 1278-1287 (2008).
  8. Erclik, T., Hartenstein, V., McInnes, R. R., Lipshitz, H. D. Eye evolution at high resolution: The neuron as a unit of homology. Developmental Biology. 332, 70-79 (2009).
  9. Wu, M., et al. Visual projection neurons in the Drosophila lobula link feature detection to distinct behavioral programs. eLife. 5, (2016).
  10. Ngo, K. T., Andrade, I., Hartenstein, V. Spatio-temporal pattern of neuronal differentiation in the Drosophila visual system: A user's guide to the dynamic morphology of the developing optic lobe. Developmental Biology. 428, 1-24 (2017).
  11. Li, X., et al. Temporal patterning of Drosophila medulla neuroblasts controls neural fates. Nature. 498, 456-462 (2013).
  12. Erclik, T., et al. Integration of temporal and spatial patterning generates neural diversity. Nature. 541, 365-370 (2017).
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  14. Suzuki, T., Kaido, M., Takayama, R., Sato, M. A temporal mechanism that produces neuronal diversity in the Drosophila visual center. Developmental Biology. 380, 12-24 (2013).
  15. Hara, Y., Sudo, T., Togane, Y., Akagawa, H., Tsujimura, H. Cell death in neural precursor cells and neurons before neurite formation prevents the emergence of abnormal neural structures in the Drosophila optic lobe. Developmental Biology. 436, 28-41 (2018).
  16. Morante, J., Erclik, T., Desplan, C. Cell migration in Drosophila optic lobe neurons is controlled by eyeless/Pax6. Development. 138, 687-693 (2011).
  17. Millard, S. S., Pecot, M. Y. Strategies for assembling columns and layers in the Drosophila visual system. Neural Development. 13, 1-17 (2018).
  18. Takemura, S. Y., Lu, Z., Meinertzhagen, I. A. Synaptic circuits of the Drosophila optic lobe: The input terminals to the medulla. Journal of Comparative Neurology. 509, 493-513 (2008).
  19. Akin, O., Bajar, B. T., Keles, M. F., Frye, M. A., Zipursky, S. L. Cell-type-Specific Patterned Stimulus-Independent Neuronal Activity in the Drosophila Visual System during Synapse Formation. Neuron. 101, 894-904 (2019).
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  28. Ting, C. Y., et al. Analyzing dendritic morphology in columns and layers. Journal of Visualized Experiments. , e55410 (2017).
  29. Özel, M. N., Langen, M., Hassan, B. A., Hiesinger, P. R. Filopodial dynamics and growth cone stabilization in Drosophila visual circuit development. eLife. 4, (2015).
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Cite this Article

Arain, U., Valentino, P., Islam, I.… More

Arain, U., Valentino, P., Islam, I. M., Erclik, T. Dissection, Immunohistochemistry and Mounting of Larval and Adult Drosophila Brains for Optic Lobe Visualization. J. Vis. Exp. (170), e61273, doi:10.3791/61273 (2021).

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