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

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Abstract

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.

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References

  • Anton S, van Loon JJ, Meijerink J, Smid HM, Takken W, Rospars JP (2003) Central projections of olfactory receptor neurons from single antennal and palpal sensilla in mosquitoes. Arthropod Struct Dev 32:319–327

    Article  PubMed  Google Scholar 

  • Arensburger P, Megy K, Waterhouse RM, Abrudan J, Amedeo P et al (2010) Sequencing of Culex quinquefasciatus establishes a platform for mosquito comparative genomics. Science 330:86–88

    Article  PubMed  CAS  Google Scholar 

  • Bohbot J, Pitts RJ, Kwon HW, Rutzler M, Robertson HM, Zwiebel LJ (2007) Molecular characterization of the Aedes aegypti odorant receptor gene family. Insect Mol Biol 16:525–537

    PubMed  CAS  Google Scholar 

  • Bello B, Holbro N, Reichert H (2007) Polycomb group genes are required for neural stem cell survival in postembryonic neurogenesis of Drosophila. Development 134:1091–1099

    Article  PubMed  CAS  Google Scholar 

  • Carey AF, Wang G, Su CY, Zwiebel LJ, Carlson JR (2010) Odorant reception in the malaria mosquito Anopheles gambiae. Nature 464:66–71

    Article  PubMed  CAS  Google Scholar 

  • Chitnis AB, Kuwada JY (1990) Axogenesis in the brain of zebrafish embryos. J Neurosci 10:1892–1905

    PubMed  CAS  Google Scholar 

  • Dacks AM, Christensen TA, Hildebrand JG (2006) Phylogeny of a serotonin-immunoreactive neuron in the primary olfactory center of the insect brain. J Comp Neurol 498:727–746

    Article  PubMed  CAS  Google Scholar 

  • Davis NT (1987) Neurosecretory neurons and their projections to the serotonin neurohemal system of the cockroach Periplaneta americana (L.), and identification of mandibular and maxillary motor neurons associated with this system. J Comp Neurol 259:604–621

    Article  PubMed  CAS  Google Scholar 

  • Eriksson BJ, Budd GE (2000) Onychophoran cephalic nerves and their bearing on our understanding of head segmentation and stem-group evolution of Arthropoda. Arthropod Struct Dev 29:197–209

    Article  PubMed  CAS  Google Scholar 

  • Feachem RG, Phillips AA, Targett GA, Snow RW (2010) Call to action: priorities for malaria elimination. Lancet 376:1517–1521

    Article  PubMed  Google Scholar 

  • Fox AN, Pitts RJ, Robertson HM, Carlson JR, Zwiebel LJ (2001) Candidate odorant receptors from the malaria vector mosquito Anopheles gambiae and evidence of down-regulation in response to blood feeding. Proc Natl Acad Sci USA 98:14693–14697

    Article  PubMed  CAS  Google Scholar 

  • Gaunt MW, Miles MA (2002) An insect molecular clock dates the origin of the insects and accords with paleontological and biogeographic landmarks. Mol Biol Evol 19:748–761

    PubMed  CAS  Google Scholar 

  • Ghaninia M, Hansson BS, Ignell R (2007a) The antennal lobe of the African malaria mosquito, Anopheles gambiae—innervation and three-dimensional reconstruction. Arthropod Struct Dev 36:23–39

    Article  PubMed  Google Scholar 

  • Ghaninia M, Ignell R, Hansson BS (2007b) Functional classification and central nervous projections of olfactory receptor neurons housed in antennal trichoid sensilla of female yellow fever mosquitoes, Aedes aegypti. Eur J Neurosci 26:1611–1623

    Article  PubMed  Google Scholar 

  • Groh C, Rössler W (2008) Caste-specific postembryonic development of primary and secondary olfactory centers in the female honeybee brain. Arthropod Struct Dev 37:459–468

    Article  PubMed  Google Scholar 

  • Halter DA, Urban J, Rickert C, Ner SS, Ito K, Travers AA, Technau GM (1995) The homeobox gene repo is required for the differentiation and maintenance of glia function in the embryonic nervous system of Drosophila melanogaster. Development 121:317–332

    PubMed  CAS  Google Scholar 

  • Hartenstein V, Spindler S, Pereanu W, Fung S (2008) The development of the Drosophila larval brain. Adv Exp Med Biol 628:1–31

    Article  PubMed  Google Scholar 

  • Harzsch S, Hansson BS (2008) Brain architecture in the terrestrial hermit crab Coenobita clypeatus (Anomura, Coenobitidae), a crustacean with a good aerial sense of smell. BMC Neurosci 9:58

    Article  PubMed  Google Scholar 

  • Hill CA, Fox AN, Pitts RJ, Kent LB, Tan PL, Chrystal MA, Cravchik A, Collins FH, Robertson HM, Zwiebel LJ (2002) G protein-coupled receptors in Anopheles gambiae. Science 298:176–178

    Article  PubMed  CAS  Google Scholar 

  • Hofbauer A, Campos-Ortega JA (1990) Proliferation pattern and early differentiation of the optic lobes in Drosophila melanogaster. Roux’s Arch Dev Biol 198:264–274

    Article  Google Scholar 

  • Ignell R, Dekker T, Ghaninia M, Hansson BS (2005) Neuronal architecture of the mosquito deutocerebrum. J Comp Neurol 493:207–240

    Article  PubMed  Google Scholar 

  • Ignell R, Hansson BS (2005) Projection patterns of gustatory neurons in the suboesophageal ganglion and tritocerebrum of mosquitoes. J Comp Neurol 492:214–233

    Article  PubMed  Google Scholar 

  • Ishii Y, Kubota K, Hara K (2005) Postembryonic development of the mushroom bodies in the ant, Camponotus japonicus. Zool Sci 22:743–753

    Article  PubMed  Google Scholar 

  • Iwai Y, Usui T, Hirano S, Steward R, Takeichi M, Uemura T (1997) Axon patterning requires DN-cadherin, a novel neuronal adhesion receptor, in the Drosophila embryonic CNS. Neuron 19:77–89

    Article  PubMed  CAS  Google Scholar 

  • Klagges BR, Heimbeck G, Godenschwege TA, Hofbauer A, Pflugfelder GO, Reifegerste R, Reisch D, Schaupp M, Buchner S, Buchner E (1996) Invertebrate synapsins: a single gene codes for several isoforms in Drosophila. J Neurosci 16:3154–3165

    PubMed  CAS  Google Scholar 

  • Kwon HW, Lu T, Rutzler M, Zwiebel LJ (2006) Olfactory responses in a gustatory organ of the malaria vector mosquito Anopheles gambiae. Proc Natl Acad Sci USA 103:13526–13531

    Article  PubMed  CAS  Google Scholar 

  • Lange AB, Orchard I, Lloyd RJ (1988) Immunohistochemical and electrochemical detection of serotonin in the nervous system of the blood-feeding bug Rhodnius prolixus. Arch Insect Biochem Physiol 8:187–201

    Article  CAS  Google Scholar 

  • Larsen C, Shy D, Spindler S, Fung S, Younossi-Hartenstein A, Hartenstein V (2009) Patterns of growth, axonal extension and axonal arborization of neuronal lineages in the developing Drosophila brain. Dev Biol 335:289–304

    Article  PubMed  CAS  Google Scholar 

  • Lu T, Qiu YT, Wang G, Kwon JY, Rutzler M, Kwon HW, Pitts RJ, van Loon JJ, Takken W, Carlson JR, Zwiebel LJ (2007) Odor coding in the maxillary palp of the malaria vector mosquito Anopheles gambiae. Curr Biol 17:1533–1544

    Article  PubMed  CAS  Google Scholar 

  • Martinez-Hernandez A, Bell KP, Norenberg MD (1977) Glutamine synthetase: glial localization in brain. Science 195:1356–1358

    Article  PubMed  CAS  Google Scholar 

  • Mayer G, Whitington PM, Sunnucks P, Pfluger HJ (2010) A revision of brain composition in Onychophora (velvet worms) suggests that the tritocerebrum evolved in arthropods. BMC Evol Biol 10:255–264

    Article  PubMed  Google Scholar 

  • Meinertzhagen IA, Hanson TE (1993) In: Bate M, Arias AM (eds) The development of the optic lobe. Cold Spring Harbor, New York, pp 1363–1491

    Google Scholar 

  • Mellanby K (1958) The alarm reaction of mosquito larvae. Entomol Exp Appl 1:153–160

    Article  Google Scholar 

  • Melo AC, Rützler M, Pitts RJ, Zwiebel LJ (2004) Identification of a chemosensory receptor from the yellow fever mosquito, Aedes aegypti, that is highly conserved and expressed in olfactory and gustatory organs. Chem Senses 29:403–410

    Article  PubMed  CAS  Google Scholar 

  • Merritt RW, Dadd RH, Walker ED (1992) Feeding behavior, natural food, and nutritional relationships of larval mosquitoes. Annu Rev Entomol 37:349–374

    Article  PubMed  CAS  Google Scholar 

  • Mysore K, Shyamala BV, Rodrigues V (2010) Morphological and developmental analysis of peripheral antennal chemosensory sensilla and central olfactory glomeruli in worker castes of Camponotus compressus (Fabricius, 1787). Arthropod Struct Dev 39:310–321

    Article  PubMed  Google Scholar 

  • Nässel DR, Elekes K (1984) Ultrastructural demonstration of serotonin immunoreactivity in the nervous system of an insect (Calliphora erythrocephala). Neurosci Lett 48:203–210

    Article  PubMed  Google Scholar 

  • Oland LA, Tolbert LP (1996) Multiple factors shape development of olfactory glomeruli: insights from an insect model system. J Neurobiol 30:92–109

    Article  PubMed  CAS  Google Scholar 

  • Olsson B, Klowden MJ (1998) Larval diet affects the alarm response of Aedes aegypti (L.) mosquitoes (Deiptera: Culicidae). J Insect Behav 11:593–596

    Article  Google Scholar 

  • Pereanu W, Younossi-Hartenstein A, Lovick J, Spindler S, Hartenstein V (2011) Lineage-based analysis of the development of the central complex of the Drosophila brain. J Comp Neurol 519:661–689

    Article  PubMed  Google Scholar 

  • Preuss U, Landsberg G, Scheidtmann KH (2003) Novel mitosis-specific phosphorylation of histone H3 at Thr11 mediated by Dlk/ZIP kinase. Nucleic Acids Res 31:878–885

    Article  PubMed  CAS  Google Scholar 

  • Rein K, Zockler M, Mader MT, Grubel C, Heisenberg M (2002) The Drosophila standard brain. Curr Biol 12:227–231

    Article  PubMed  CAS  Google Scholar 

  • Rodrigues V, Hummel T (2008) Development of the Drosophila olfactory system. Adv Exp Med Biol 628:82–101

    Article  PubMed  Google Scholar 

  • Siju KP, Hansson BS, Ignell R (2008) Immunocytochemical localization of serotonin in the central and peripheral chemosensory system of mosquitoes. Arthropod Struct Dev 37:248–259

    Article  PubMed  CAS  Google Scholar 

  • Snow PM, Patel NH, Harrelson AL, Goodman CS (1987) Neural-specific carbohydrate moiety shared by many surface glycoproteins in Drosophila and grasshopper embryos. J Neurosci 7:4137–4144

    PubMed  CAS  Google Scholar 

  • Spindler SR, Hartenstein V (2010) The Drosophila neural lineages: a model system to study brain development and circuitry. Dev Genes Evol 220:1–10

    Article  PubMed  Google Scholar 

  • Sprecher SG, Cardona A, Hartenstein V (2011) The Drosophila larval visual system: high-resolution analysis of a simple visual neuropil. Dev Biol. doi:10.1016/j.ydbio.2011.07.006

  • van der Hel WS, Notenboom RG, Bos IW, van Rijen PC, van Veelen CW, de Graan PN (2005) Reduced glutamine synthetase in hippocampal areas with neuron loss in temporal lobe epilepsy. Neurology 64:326–333

    Article  PubMed  Google Scholar 

  • van Haeften T, Schooneveld H (1993) Diffuse serotoninergic neurohemal systems associated with cerebral and subesophageal nerves in the head of the Colorado potato beetle Leptinotarsa decemlineata. Cell Tissue Res 273:327–333

    Article  Google Scholar 

  • Wang G, Carey AF, Carlson JR, Zwiebel LJ (2010) Molecular basis of odor coding in the malaria vector mosquito Anopheles gambiae. Proc Natl Acad Sci USA 107:4418–4423

    Article  PubMed  CAS  Google Scholar 

  • Ward MM, Jobling AI, Puthussery T, Foster LE, Fletcher EL (2004) Localization and expression of the glutamate transporter, excitatory amino acid transporter 4, within astrocytes of the rat retina. Cell Tissue Res 315:305–310

    Article  PubMed  CAS  Google Scholar 

  • Yeates DK, Wiegmann BM (1999) Congruence and controversy: toward a higher-level phylogeny of Diptera. Annu Rev Entomol 44:397–428

    Article  PubMed  CAS  Google Scholar 

  • Zdobnov EM, von Mering C, Letunic I, Torrents D, Suyama M, Copley RR, Christophides GK, Thomasova D, Holt RA, Subramanian GM, Mueller HM, Dimopoulos G, Law JH, Wells MA, Birney E, Charlab R, Halpern AL, Kokoza E, Kraft CL, Lai Z, Lewis S, Louis C, Barillas-Mury C, Nusskern D, Rubin GM, Salzberg SL, Sutton GG, Topalis P, Wides R, Wincker P, Yandell M, Collins FH, Ribeiro J, Gelbart WM, Kafatos FC, Bork P (2002) Comparative genome and proteome analysis of Anopheles gambiae and Drosophila melanogaster. Science 298:149–159

    Article  PubMed  CAS  Google Scholar 

  • Zipursky SL, Venkatesh TR, Teplow DB, Benzer S (1984) Neuronal development in the Drosophila retina: monoclonal antibodies as molecular probes. Cell 36:15–26

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We are very grateful to GM Technau, JB Skeath, CY Lee, J Pielage, and the Developmental Studies Hybridoma Bank (DSHB) for the antibodies. We also thank all the members of the Reichert lab for discussions and assistance. We are very thankful to the two unknown reviewers for their valuable comments on the previous version of the manuscript, their constructive criticism have greatly improved the manuscript. We are very grateful to Danica Jančariová and Mohamad Sater at Vector Control Centre, STPHI, Basel, Switzerland for insect rearing. This work was supported by the Indo-Swiss Joint Research Project (ISJRP).

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Authors and Affiliations

  1. Biozentrum, University of Basel, Klinglebergstrasse 50, CH-4056, Basel, Switzerland

    Keshava Mysore, Susanne Flister & Heinrich Reichert

  2. Swiss Tropical and Public Health Institute, Socinstrasse 57, CH-4002, Basel, Switzerland

    Pie Müller

  3. University of Basel, Petersplatz 1, CH-4003, Basel, Switzerland

    Pie Müller

  4. National Centre for Biological Sciences, Tata Institute for Fundamental Research, GKVK Campus, Bellary Road, Bangalore, 560065, India

    Veronica Rodrigues

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  1. Keshava Mysore
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  2. Susanne Flister
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  4. Veronica Rodrigues
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  5. Heinrich Reichert
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Correspondence to Heinrich Reichert.

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Communicated by V. Hartenstein

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Supp. Fig. 1

Cross-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 (b–e). 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)

Supp. Fig. 2

Cross-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)

Supp. Fig. 3

Cross-reactivity of antibodies against the pupal (24 h APF) brain of A. aegypti. Cortical and neuropil regions are labeled by anti-tubulin antibodies (a–d). 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)

Supp. Fig. 4

Cross-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)

Supp. Fig. 5

Cross-reactivity of antibodies against the adult brain of A. aegypti. Labeling by anti-HRP (a) and anti-tubulin antibodies (b–e) 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)

Supp. Fig. 6

Cross-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)

Supplementary Table 1

Antibodies 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)

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Mysore, K., Flister, S., Müller, P. et al. Brain development in the yellow fever mosquito Aedes aegypti: a comparative immunocytochemical analysis using cross-reacting antibodies from Drosophila melanogaster . Dev Genes Evol 221, 281–296 (2011). https://doi.org/10.1007/s00427-011-0376-2

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