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Ontogeny of pioneer neurons in the antennal nervous system of the grasshopper Schistocerca gregaria

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Abstract

The nervous system of the antenna of the grasshopper Schistocerca gregaria consists of two nerve tracts in which sensory cells project their axons to the brain. Each tract is pioneered early in embryogenesis by a pair of identified cells located apically in the antennal lumen. The pioneers are thought to originate in the epithelium of the antenna and then delaminate into the lumen where they commence axogenesis. However, unambiguous molecular identification of these cells in the epithelium, of an identifiable precursor, and of their mode of generation has been lacking. In this study, we have used immunolabeling against neuron-specific horseradish peroxidase and against Lachesin, a marker for differentiating epithelial cells, in combination with the nuclear stain DAPI, to identify the pioneers within the epithelium of the early embryonic antenna. We then track their delamination into the lumen as differentiated neurons. The pioneers are not labeled by the mesodermal/mesectodermal marker Mes3, consistent with an epithelial (ectodermal) origin. Intracellular dye injection, as well as labeling against the mitosis marker phospho-histone 3, identifies precursor cells in the epithelium, each associated with a column of cells. Culturing with the S-phase label 5-ethynyl-2′-deoxyuridine (EdU) shows that both a precursor and its column have incorporated the label, confirming a lineage relationship. Each set of pioneers can be shown to belong to a separate lineage of such epithelial cells, and the precursors remain and are proliferative after generating the pioneers. Analyses of mitotic spindle orientation then enable us to propose a model in which a precursor generates its pioneers asymmetrically via self-renewal.

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Acknowledgements

We thank Drs. Sanchez and Ganfornina for the gift of the Lazarillo antibody, Dr. Goodman for the gift of the Mes3 antibody, and Dr. Bastiani for the gift of the Lachesin antibody. We thank Dr. Yu Liu for the critical reading of the manuscript and Karin Fischer for the excellent technical assistance.

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Authors and Affiliations

  1. Graduate School of Systemic Neuroscience, Biocenter, Ludwig-Maximilians-Universität München, Grosshadernerstrasse 2, Martinsried, 82152, Planegg, Germany

    George Boyan & Erica Ehrhardt

  2. Janelia Farm Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, VA, 20147,, USA

    Erica Ehrhardt

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  1. George Boyan
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  2. Erica Ehrhardt
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Correspondence to George Boyan.

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All experiments were performed in accordance with the guidelines for animal welfare as laid down by the Deutsche Forschungsgemeinschaft.

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The authors declare that they have no competing interests.

Funding

Both authors received support for this study from the Graduate School of Systemic Neuroscience, University of Munich.

Additional information

Communicated by Claude Desplan

Both authors contributed equally to this study.

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Boyan, G., Ehrhardt, E. Ontogeny of pioneer neurons in the antennal nervous system of the grasshopper Schistocerca gregaria . Dev Genes Evol 227, 11–23 (2017). https://doi.org/10.1007/s00427-016-0565-0

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