Development Genes and Evolution

, Volume 221, Issue 5, pp 281–296

Brain development in the yellow fever mosquito Aedes aegypti: a comparative immunocytochemical analysis using cross-reacting antibodies from Drosophila melanogaster

  • Keshava Mysore
  • Susanne Flister
  • Pie Müller
  • Veronica Rodrigues
  • Heinrich Reichert
Original Article

DOI: 10.1007/s00427-011-0376-2

Cite this article as:
Mysore, K., Flister, S., Müller, P. et al. Dev Genes Evol (2011) 221: 281. doi:10.1007/s00427-011-0376-2


Considerable effort has been directed towards understanding the organization and function of peripheral and central nervous system of disease vector mosquitoes such as Aedes aegypti. To date, all of these investigations have been carried out on adults but none of the studies addressed the development of the nervous system during the larval and pupal stages in mosquitoes. Here, we first screen a set of 30 antibodies, which have been used to study brain development in Drosophila, and identify 13 of them cross-reacting and labeling epitopes in the developing brain of Aedes. We then use the identified antibodies in immunolabeling studies to characterize general neuroanatomical features of the developing brain and compare them with the well-studied model system, Drosophila melanogaster, in larval, pupal, and adult stages. Furthermore, we use immunolabeling to document the development of specific components of the Aedes brain, namely the optic lobes, the subesophageal neuropil, and serotonergic system of the subesophageal neuropil in more detail. Our study reveals prominent differences in the developing brain in the larval stage as compared to the pupal (and adult) stage of Aedes. The results also uncover interesting similarities and marked differences in brain development of Aedes as compared to Drosophila. Taken together, this investigation forms the basis for future cellular and molecular investigations of brain development in this important disease vector.


Aedes aegypti Fruitfly Brain development Optic lobes Subesophageal ganglion 

Supplementary material

427_2011_376_MOESM1_ESM.pdf (1.5 mb)
Supp. Fig. 1Cross-reactivity of antibodies against the larval brain (stage L4) of A. aegypti. Information on the overall organization of the larval brain is provided by the anti-HRP antibody (a) and by the 4 anti-tubulin antibodies (be). Anti-22 C10 immunostaining reveal comparable aspects of the larval brain both in cortical and neuropil regions (f). Scale bar 100 μm (PDF 3,379 kb; PDF 1524 kb)
427_2011_376_MOESM2_ESM.pdf (1.3 mb)
Supp. Fig. 2Cross-reactivity of antibodies against the larval brain (stage L4) of A. aegypti. Anti-3 C11 (a) and anti-nc82 (b) immunostaining reveals a complex compartmentlike organization in the neuropil of the supraesophageal and subesophageal ganglia. Anti-5HT immunostaining (c) labels the small subset of 5HT immunoreactive cell bodies as well as their projections in the neuropil, commissures, and connectives of the supraesophageal and subesophageal ganglia. Anti-GS and anti-Repo immunolabeling (d, e) labels glia that are associated with the cortical regions and the neuropil regions of the. Mitotic activity throughout the larval brain is indicated by anti-PH3 immunostaining (f). Scale bar 100 μm (PDF 2,771 kb; PDF 1,306 kb)
427_2011_376_MOESM3_ESM.pdf (1.7 mb)
Supp. Fig. 3Cross-reactivity of antibodies against the pupal (24 h APF) brain of A. aegypti. Cortical and neuropil regions are labeled by anti-tubulin antibodies (ad). A similar but more detailed labeling is seen in anti-HRP immunostained preparations (e) while anti-22 C10 antibody labels more discrete axon tracts and fascicles in the brain rather than compact neuropil domains (f). Yellow dots highlight the antennal lobe in all the cases. Scale bar 100 μm (PDF 3,816 kb; PDF 1,756 kb)
427_2011_376_MOESM4_ESM.pdf (1.5 mb)
Supp. Fig. 4Cross-reactivity of antibodies against the pupal (24 h APF) brain of A. aegypti. More precise information on the organization of the neuropils is obtained by anti-3 C11 (a), anti-nc82 (b) and anti-N-Cad immunostaining (c). Whereas a restricted regionalized labeling is seen in anti-5HT immunostained pupal brains (d). Glial cell bodies are immunolabeled with anti-GS antibody (e) and as well anti-repo immunolabeling (f). Yellow dots highlight the antennal lobe in all the cases. Scale bar 100 μm (PDF 3,302 kb; PDF 1,556 kb)
427_2011_376_MOESM5_ESM.pdf (1.7 mb)
Supp. Fig. 5Cross-reactivity of antibodies against the adult brain of A. aegypti. Labeling by anti-HRP (a) and anti-tubulin antibodies (be) reveals general features of the adult brain. More specific immunolabeling of neuropil regions is observed with anti-22 C10 (f) antibody staining. Yellow dots highlight the antennal lobe in all the cases. Scale bar 100 μm (PDF 3,929 kb; PDF 1,772 kb)
427_2011_376_MOESM6_ESM.pdf (1.6 mb)
Supp. Fig. 6Cross-reactivity of antibodies against the adult brain of A. aegypti. More specific immunolabeling of neuropil regions with anti-3 C11 (a), anti-nc82 (b) and anti-DN Cadherin (c) also reveals features which are very similar to the pupal brain Immunostaining with the anti-5HT (d) antibody, anti-GS (e), and anti-Repo (f) antibodies labels serotonergic neurons and glial cells respectively in the adult brain. Yellow dots highlight the antennal lobe in all the cases. Scale bar 100 μm (PDF 3,551 kb; PDF 1,633 kb)
427_2011_376_MOESM7_ESM.pdf (3.6 mb)
Supplementary Table 1Antibodies screened against the mosquito brain for cross-reactivity. 30 antibodies from frutifly and other sources were screened for cross-reactivity against the developing brain of the mosquito Aedes aegypti. The cross-reacting antibodies are indicated by positive sign and the antibodies that do not cross react are indicated by negative sign. CYL-Cheng-Yu Lee, GT-Gerhard Technau, JS-James Skeath, VR-Veronica Rodrigues (PDF 3,695 kb; PDF 3,695 kb)

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Keshava Mysore
    • 1
  • Susanne Flister
    • 1
  • Pie Müller
    • 2
    • 3
  • Veronica Rodrigues
    • 4
  • Heinrich Reichert
    • 1
  1. 1.BiozentrumUniversity of BaselBaselSwitzerland
  2. 2.Swiss Tropical and Public Health InstituteBaselSwitzerland
  3. 3.University of BaselBaselSwitzerland
  4. 4.National Centre for Biological SciencesTata Institute for Fundamental Research, GKVK CampusBangaloreIndia