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Ex vivo observation of insect hemocyte behavior against beads and nematodes in the presence of insect plasma

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Abstract

Recognition of foreign targets by insect hemocytes is a crucial first step for insect immunity against invading multicellular organisms in the hemocoel. To understand the mechanism of recognition, we observed the hemocyte behavior of Galleria mellonella Linnaeus (Lepidoptera: Pyralidae) larvae against beads and the nonparasitic nematode Caenorhabditis elegans (Maupas) (Rhabditida: Rhabditidae) in the presence of plasma ex vivo using time-lapse microscopy. Both granular cells and plasmatocytes adhered to and spread on the surface of beads and nematodes. In addition, the spread plasmatocytes actively moved over the beads and nematodes. These results suggest that not only granular cells but also plasmatocytes can recognize foreign targets in the presence of insect plasma and that spread plasmatocytes can actively search for foreign targets. Hemocyte adhesion to beads and nematodes ex vivo was similar to that of the in vivo 1 h after injection. A divalent cation chelator inhibited the spreading and adhesion of plasmatocytes ex vivo, but it did not affect the adhesion of granular cells. The present method enables the analysis of acute hemocyte response against foreign targets in the presence of plasma.

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Acknowledgements

The authors would like to thank the Caenorhabditis Genetics Center for providing nematode strain and Enago (www.enago.jp) for the English language review of the manuscript. This work was supported by a Grant-in-Aid for Challenging Exploratory Research (T.Y., Grant Number 23658051) and a Grant-in-Aid for Scientific Research (C) (T.Y, Grant Number 18K05676).

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Authors and Affiliations

  1. Laboratory of Nematology, Department of Applied Biological Sciences, Faculty of Agriculture, Saga University, Saga, 840-8502, Japan

    Masaya Ono, Chisato Arimatsu & Toyoshi Yoshiga

  2. United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Japan

    Masaya Ono & Toyoshi Yoshiga

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  1. Masaya Ono
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  2. Chisato Arimatsu
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  3. Toyoshi Yoshiga
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Correspondence to Toyoshi Yoshiga.

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Movie 1. Hemocyte behavior on a Dowex bead. The focus is on the hemocytes on the top of the bead. Granular cells (black arrowheads) and plasmatocytes (white arrowheads) spread soon after descending onto the bead and plasmatocytes actively move over the bead. (MP4 5241 kb)

Movie 2. Hemocyte behavior on a nematode. Granular cells (black arrowheads) remained stationary at the places, where they adhered, but the spread plasmatocytes (white arrowheads) actively moved over the nematode. (MP4 6761 kb)

Movie 3. Hemocyte behavior on a nematode in the presence of EDTA. While some granular cells adhered on the nematode, no spreading or adhesion of plasmatocytes was observed. The black arrowheads indicate granular cells. (MP4 8076 kb)

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Ono, M., Arimatsu, C. & Yoshiga, T. Ex vivo observation of insect hemocyte behavior against beads and nematodes in the presence of insect plasma. Appl Entomol Zool 55, 65–70 (2020). https://doi.org/10.1007/s13355-019-00654-w

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  • DOI: https://doi.org/10.1007/s13355-019-00654-w

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