Eur. J. Entomol. 115: 157-166, 2018 | DOI: 10.14411/eje.2018.015
Molecular cloning and functional analyses of an adhesion molecule, neuroglian, in Mythimna separata (Lepidoptera: Noctuidae)Original article
- Applied Entomology, Graduate School of Bio-Agricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan; e-mails: yokoi123@affrc.go.jp, m____sssuzuki@gmail.com, k-miura@agr.nagoya-u.ac.jp
Insect cellular immune reaction, which generally includes phagocytosis, encapsulation and nodule formation, is achieved by hemocytes circulating in insect haemolymph. The shift of hemocytes from the normal phase to the adhering phase is an important process in the cellular immune reaction, which includes the attachment of hemocytes to foreign surfaces or other hemocytes via adhesion factors. Neuroglian is one of the adhering factors associated with encapsulation in Manduca sexta and Drosophila melanogaster. Here we studied the localization of neuroglian (MsNrg) in Mythimna separata and its functional role in the cellular immune reaction. The distribution of MsNrg mRNA between hemocyte populations was determined using real-time quantitative reverse transcription PCR and in situ hybridization, which revealed that MsNrg was highly expressed in adhering hemocytes, especially in plasmatocytes. Unexpectedly, the transcript was observed as well in non-adhering hemocytes, implying neuroglian has a function in non-adhering hemocytes. Moreover, we showed that the amount of MsNrg mRNA was not changed by injections of either biotic or abiotic non-selves. Fewer latex beads were fully encapsulated by hemocytes in larvae treated with MsNrg double-stranded RNA than in control larvae, but their ability to achieve phagocytosis and nodule formation remained unchanged by the MsNrg knockdown. These results indicate that the function of neuroglian in the cellular immune reaction is conserved in D. melanogaster and lepidopteran species, and neuroglian in non-adhering hemocytes could possess unidentified function.
Keywords: Lepidoptera, Noctuidae, Mythimna separata, neuroglian, hemocyte adhesion, encapsulation, in situ hybridization, real-time quantitative reverse transcript PCR, RNA interference
Received: February 7, 2018; Revised: March 15, 2018; Accepted: March 15, 2018; Published online: April 10, 2018 Show citation
| ACS | AIP | APA | ASA | Harvard | Chicago | Chicago Notes | IEEE | ISO690 | MLA | NLM | Turabian | Vancouver |
References
- Beaudoing E., Freier S., Wyatt J.R., Claverie J.M. & Gautheret D. 2010: Patterns of variant polyadenylation signal usage in human genes. - Genome Res. 10: 1001-1010.
Go to original source... - Bieber A.J., Snow P.M., Hortsch M., Patel N.H., Jacobs J.R., Traquina Z.R., Schilling J. & Goodman C.S. 1989: Drosophila neuroglian: a member of the immunoglobulin superfamily with extensive homology to the vertebrate neural adhesion molecule L1. - Cell 59: 447-460.
Go to original source... - Chen C.L., Lampe D.J., Robertson H.M. & Nardi J.B. 1997: Neuroglian is expressed on cells destined to form the prothoracic glands of Manduca embryos as they segregate from surrounding cells and rearrange during morphogenesis. - Dev. Biol. 181: 1-13.
Go to original source... - Da Silva J.B., De Albuquerque C.M., De Araujo E.C., Peixoto C.A. & Hurd H. 2000: Immune defense mechanisms of Culex quinquefasciatus (Diptera: Culicidae) against Candida albicans infection. - J. Invertebr. Pathol. 76: 257-262.
Go to original source... - Dean P., Richards E.H., Edwards J.P., Reynolds S.E. & Charnley K. 2004: Microbial infection causes the appearance of haemocytes with extreme spreading ability in monolayers of the tobacco hornworm Manduca sexta. - Dev. Comp. Immunol. 28: 689-700.
Go to original source... - Eleftherianos I., Gökçen F., Felföldi G., Millichap P.J., Trenczek T.E. French-Constant R.H. & Reynolds S.E. 2007: The immunoglobulin family protein Hemolin mediates cellular immune responses to bacteria in the insect Manduca sexta. - Cell Microbiol. 5: 1137-1147.
Go to original source... - Faraldo A.C., Gregorio E.A. & Lello E. 2008: Morphological and quantitative aspects of nodule formation in hemolymph of the blowfly Chrysomya megacephala (Fabricius, 1794). - Exp. Parasitol. 118: 372-377.
Go to original source... - Hall S.G. & Bieber A.J. 1997: Mutations in the Drosophila neuroglian cell adhesion molecule affect motor neuron pathfinding and peripheral nervous system patterning. - J. Neurobiol. 32: 325-340.
Go to original source... - Hernandez S., Lanz H., Rodriguez M.H., Torres J.A., Martinez-Palomo A. & Tsutsumi V. 1999: Morphological and cytochemical characterization of female Anopheles albimanus (Diptera: Culicidae) hemocytes. - J. Med. Entomol. 36: 426-434.
Go to original source... - Hortsch M. 1996: The L1 family of neural cell adhesion molecules: Old proteins performing new tricks. - Neuron 17: 587-593.
Go to original source... - Hortsch M., Wang Y.M., Marikar Y. & Bieber A.J. 1995: The cytoplasmic domain of the Drosophila cell adhesion molecule neuroglian is not essential for its homophilic adhesive properties in S2 cells. - J. Biol. Chem. 270: 18809-18817.
Go to original source... - Kato Y., Yoshida T., Miura K., Tanaka T., Nakamatsu Y. & Ochiai M. 2014: A novel type of hemocytes localizing melanization with high-spreading behavior in Mythimna separata. - Arch. Insect Biochem. Physiol. 86: 220-239.
Go to original source... - Lavine M.D. & Strand M.R. 2002: Insect hemocytes and their role in immunity. - Insect Biochem. Mol. Biol. 32: 1295-1309.
Go to original source... - Lemaitre B. & Hoffmann J. 2007: The host defense of Drosophila melanogaster. - Annu. Rev. Immunol. 47: 697-743.
Go to original source... - Marchler-Bauer A., Bo Y., Han L., He J., Lanczycki C.J., Lu S., Chitsaz F., Derbyshire M.K., Geer R.C., Gonzales N.R. et al. 2017: CDD/SPARCLE: Functional classification of proteins via subfamily domain architectures. - Nucl. Acids Res. 45: D200-D203.
Go to original source... - Marmaras V.J. & Lampropoulou M. 2009: Regulators and signalling in insect hemocyte immunity. - Cell Signal. 21: 186-195.
Go to original source... - Nakamatsu Y., Kuriya K., Harvey J.A. & Tanaka T. 2006: Influence of nutrient deficiency caused by host developmental arrest on the growth and development of a koinobiont parasitoid. - J. Insect Physiol. 52: 1105-1112.
Go to original source... - Nardi J.B. 1994: Rearrangement of epithelial cell types in an insect wing monolayer is accompanied by differential expression of a cell surface protein. - Dev. Dyn. 199: 315-325.
Go to original source... - Nardi J.B., Pilas B., Ujhelyi E., Garsha K. & Kanost M.R. 2003: Hematopoietic organs of Manduca sexta and hemocyte lineages. - Dev. Genes Evol. 213: 477-491.
Go to original source... - Nardi J.B., Zhuang S., Pilas B., Bee C.M. & Kanost M.R. 2005: Clustering of adhesion receptors following exposure of insect blood cells to foreign surfaces. - J. Insect Physiol 51: 555-564.
Go to original source... - Nardi J.B., Pilas B., Bee C.M., Zhuang S., Garsha K. & Kanost M.R. 2006: Neuroglian-positive plasmatocytes of Manduca sexta and the initiation of hemocyte attachment to foreign surfaces. - Dev. Comp. Immunol. 30: 447-462.
Go to original source... - Siegenthaler D., Enneking E.M., Moreno E. & Pielage J. 2015: L1CAM/Neuroglian controls the axon-axon interactions establishing layered and lobular mushroom body architecture. - J. Cell Biol. 208: 1003-1018.
Go to original source... - Sun S.C., Lindstrom I., Boman H.G., Faye I. & Schmidt O. 1990: Hemolin: An insect-immune protein belonging to the immunoglobulin superfamily. - Science 250: 1729-1732.
Go to original source... - Suzuki M. & Tanaka T. 2007: Development of Meteorus pulchricornis and regulation of its noctuid host, Pseudaletia separata. - J. Insect Physiol. 53: 1072-1078.
Go to original source... - Suzuki M., Miura K. & Tanaka T. 2008: The virus-like particles of a braconid endoparasitoid wasp, Meteorus pulchricornis, inhibit hemocyte spreading in its noctuid host, Pseudaletia separata. - J. Insect Physiol. 54: 1015-1022.
Go to original source... - Suzuki M., Miura K. & Tanaka T. 2009: Effects of the virus-like particles of a braconid endoparasitoid, Meteorus pulchricornis, on hemocytes and hematopoietic organs of its noctuid host, Pseudaletia separata. - Appl. Entomol. Zool. 44: 115-125.
Go to original source... - Watson F.L., Puttmann-Holgado R., Thomas F., Lamar D.L., Hughes M., Kondo M., Rebel V.I. & Schmucker D. 2005: Extensive diversity of Ig-superfamily proteins in the immune system of insects. - Science 309: 1874-1878.
Go to original source... - Wiegand C., Levin D., Gillespie J., Willott E., Kanost M.R. & Trenczek T. 2000: Monoclonal antibody MS13 identifies a plasmatocyte membrane protein and inhibits encapsulation and spreading reactions of Manduca sexta hemocytes. - Arch. Insect Biochem. Physiol. 45: 95-108.
Go to original source... - Williams M.J. 2009: The Drosophila cell adhesion molecule Neuroglian regulates Lissencephaly-1 localisation in circulating immunosurveillance cells. - BMC Immunol. 10: 17, 12 pp.
Go to original source... - Zhuang S., Kelo L., Nardi J.B. & Kanost M.R. 2007: Neuroglian on hemocyte surfaces is involved in homophilic and heterophilic interactions of the innate immune system of Manduca sexta. - Dev. Comp. Immunol 31: 1159-1167.
Go to original source...
This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (CC BY 4.0), which permits use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.




