DOI QR코드

DOI QR Code

Exosome isolation from hemolymph of Korean rhinoceros beetle, Allomyrina dichotoma (Coleoptera: Scarabaeidae)

  • KIM, Hong Geun (Industrial Insect Division, National Institute of Agricultural Science, Rural Development Administration) ;
  • KWON, Kisang (Department of Biomedical Laboratory Science, College of Health and Welfare, Kyungwoon University) ;
  • SUH, Hyun-Woo (Department of Anatomy, College of Medicine, Chungnam National University) ;
  • LEE, Seokhyun (Industrial Insect Division, National Institute of Agricultural Science, Rural Development Administration) ;
  • PARK, Kwan-Ho (Industrial Insect Division, National Institute of Agricultural Science, Rural Development Administration) ;
  • KWON, O-Yu (Department of Anatomy, College of Medicine, Chungnam National University) ;
  • CHOI, Ji-Young (Industrial Insect Division, National Institute of Agricultural Science, Rural Development Administration)
  • Received : 2015.08.27
  • Accepted : 2015.09.24
  • Published : 2015.11.29

Abstract

Exosomes are 30-150 nm vesicles that are secreted from a range of cells. Recently, exosomes have been the subject of considerable research because there is mounting awareness of their diverse functions, including a role in cell-cell communication and presenting pathogens for immune responses. Exosomes contain diverse nucleic acid and protein cargos, derived not only from the organism but also from pathogens, making them suitable for use in disease diagnosis. The Korean rhinoceros beetle, Allomyrina dichotoma (Coleoptera: Scarabaeidae), is commercially reared in Korea for the pet trade and is used in traditional medicine for liver-related diseases. However, several insect diseases caused by bacteria, fungi and viruses have been reported in A. dichotoma mass-rearing facilities. Identifying these diseases with accuracy and in a timely manner is of paramount importance. Such diagnosis can be accomplished by identifying the nucleic acid or amino acid fragments from these disease-causing pathogens in the exosome of A. dichotoma. We isolated exosomes from the hemolymph of A. dichotoma and used them to analyze exosome RNA and proteins. We confirmed the isolation of exosomes through RNA profiling, protein analysis and Western blotting. Our research established a solid foundation for using insect exosome protein and RNA analyses for the accurate diagnosis of insect diseases. To our knowledge, this is the first report of exosome isolation from insect hemolymph.

Keywords

Acknowledgement

Supported by : Rural Development Administration

References

  1. Aalberts M, Sostaric E, Wubbolts R et al. (2013) Spermatozoa recruit prostasomes in response to capacitation induction. Biochimica et Biophysica Acta 1834: 2326-2335. https://doi.org/10.1016/j.bbapap.2012.08.008
  2. Andrulis E, Werner J, Nazarian A, Erdjumnet-Bromage H, Tempst P, Lis JT (2002) The RNA processing exosome is linked to elongating RNA polymerase II in Drosophila. Nature 420: 837-841. https://doi.org/10.1038/nature01181
  3. Chlebowshi A, Lubas M, Jensen TH, Dziembowski A (2013) RNA decay machines: the exosome. Biochimica et Biophysica Acta (BBA)-Gene Regulatory Mechanisms 1829: 552-560. https://doi.org/10.1016/j.bbagrm.2013.01.006
  4. Choi YH, Lee KY, Yang KM, Jeong YM, Seo JS (2006) Effect of larva extract of Allomyrina dichotoma on carbon tetrachloride-induced hepatotoxicity in mice. Journal of the Korean Society of Food Science and Nutrition 35: 1349-1355. https://doi.org/10.3746/jkfn.2006.35.10.1349
  5. Chung MY, Yoon YI, Hwang JS, Goo TW, Yun EY (2014) Anti-obesity effect of Allomyrina dichotoma (Arthropoda: Insecta) larvae ethanol extract on 3T3-L1 adipocyte differentiation. Entomological Research 44: 9-16. https://doi.org/10.1111/1748-5967.12044
  6. Conde-Vancells J, Rodriguez-Suarez E, Embade N et al. (2008) Characterization and comprehensive proteome profiling of exosomes secreted by hepatocytes. Journal of Proteome Research 7: 5157-5166. https://doi.org/10.1021/pr8004887
  7. Corrigan L, Redhai S, Leiblich A et al. (2014) BMP-regulated exosomes from Drosophila male reproductive glands reprogram female behavior. Journal of Cell Biology 206: 671-688. https://doi.org/10.1083/jcb.201401072
  8. Denzer K, Kleijmeer MJ, Heijnen HFG, Stoorvogle W, Geuze HJ (2000) Exosome: from internal vesicle of the multivesicular body to intercellular signaling device. Journal of Cell Science 113: 3365-3374.
  9. Gloeckner CJ, Boldt K, Schumacher A, Roepman R, Ueffing M (2007) A novel tandem affinity purification strategy for the efficient isolation and characterization of native protein complexes. Proteomics 7: 4228-4234. https://doi.org/10.1002/pmic.200700038
  10. Graham AC, Kiss DL, Andrulis ED (2006) Differential distribution of exosome subunits at the nuclear lamina and in cytoplasmic foci. Molecular Biology of the Cell 17: 1399-1409. https://doi.org/10.1091/mbc.e05-08-0805
  11. Hessle V (2011) Characterization of RNA exosome in insect cells: role in mRNA surveillance (Dissertation). Stockholm University, Stockholm, Sweden.
  12. Kim DS, Huh J, You GC et al. (2007) Allomyrina dichotoma larva extracts protect streptozotocin-induced oxidative cytotoxicity. Environmental Health and Toxicology 22: 349-355.
  13. Kim M, Youn K, Yun EY et al. (2014) Effects of solvent fractions of Allomyrina dichotoma larvae through the inhibition of in vitro BACE1 and ${\beta}$-amyloid (25-35)-induced toxicity in rat pheochromocytoma PC12 cells. Entomological Research 44: 23-30. https://doi.org/10.1111/1748-5967.12046
  14. Koles K, Budnik V (2012) Exosomes go with the Wnt. Cellular Logistics 2: 169-173. https://doi.org/10.4161/cl.21981
  15. Kosaka N, Yoshioka Y, Katsuda T, Ono M, Ochiya T (2014) Exosome in disease biology, diagnosis, and therapy. Inflammation and Regeneration 34: 233-239. https://doi.org/10.2492/inflammregen.34.233
  16. Lane RE, Korbie D, Anderson W, Vaidyanathan R, Trau M (2015) Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing. Scientific Reports 5: 7639. doi:10.1038/srep07639
  17. Lee S, Park KH, Nam SH, Kwak KW, Choi JY (2015) First report of Oryctes rhinoceros nudivirus (Coleoptera: Scarabaeidae) causing severe disease in Allomyrina dichotoma in Korea. Journal of Insect Science 15: 26. doi:10.1093/jisesa/iev002
  18. Lotvall J, Valadi H (2007) Cell to cell signaling via exosomes through esRNA. Cell Adhesion & Migration 1: 156-158. https://doi.org/10.4161/cam.1.3.5114
  19. Lykke-Andersen S, Brodersen DE, Jensen TH (2009) Origins of the eukaryotic exosomes. Journal of Cell Science 122: 1487-1494. https://doi.org/10.1242/jcs.047399
  20. Mitchell P, Tollervey D (2003) An NMD pathway in yeast involving accelerated deadenylation and exosome-mediated 3′ . . . >5′ degradation. Molecular Cell 11: 1405-1413. https://doi.org/10.1016/S1097-2765(03)00190-4
  21. Miyanoshita A, Hara S, Sugiyama M et al. (1996) Isolation and characterization of a new member of the insect defensing family from a beetle, Allomyrina dichotoma. Biochemistry Biophysics Research Communication 220: 526-531. https://doi.org/10.1006/bbrc.1996.0438
  22. Palmerini CA, Carlini E, Nicolucci A, Arienti G (1999) Increase of human spermatozoa intracellular $Ca^{2+}$ concentration after fusion with prostasomes. Cell Calcium 25: 291-296. [Cited 25 Aug 2015.] Available from URL: http://dx.doi.org/10.1054/ceca.1999.0031.
  23. Rabinowits G, Gercel-Taylor C, Day JM, Taylor DD, Kloecker GH (2009) Exosomal microRNA: a diagnostic marker for lung cancer. Clinical Lung Cancer 10: 42-46. https://doi.org/10.3816/CLC.2009.n.006
  24. Sagisaka A, Miyanoshita A, Ishibashi J, Yamakawa M (2001) Purification, characterization and gene expression of a glycine and proline-rich antibacterial protein family from larvae of a beetle, Allomyrina dichotoma. Insect Molecular Biology 10: 293-302. https://doi.org/10.1046/j.0962-1075.2001.00261.x
  25. Simpson RJ, Lim JWE, Moritz RL, Mathivanan S (2009) Exosomes: proteomic insights and diagnostic potential. Expert Review of Proteomics 6: 267-283. https://doi.org/10.1586/epr.09.17
  26. Sullivan R, Saez F (2013) Epididymosomes, prostasomes, and liposomes: their roles in mammalian male reproductive physiology. Reproduction (Cambridge, England) 146: R21-R35. https://doi.org/10.1530/REP-13-0058
  27. Szatanek R, Baran J, Siedlar M, Baj-Krzyworzeka M (2015) Isolation of extracellular vesicles: determining the correct approach (Review). International Journal of Molecular Medicine 36: 11-17. https://doi.org/10.3892/ijmm.2015.2194
  28. Taylor DD, Gercel-Taylor C (2014) MicroRNA signatures of tumor-derived exosomes as diagnostic biomarkers of ovarian cancer. Gynecologic Oncology 110: 13-21.
  29. van Niel G, Porto-Carreiro I, Simoes S, Raposo G (2006) Exosomes: a common pathway for a specialized function. Journal of Biochemistry 140: 13-21. https://doi.org/10.1093/jb/mvj128
  30. Vlassov AV, Magdaleno S, Setterquist R, Conrad R (2012) Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. Biochimica et Biophysica Acta (BBA)-General Subjects 1820: 940-948. https://doi.org/10.1016/j.bbagen.2012.03.017
  31. Winnebeck EC, Millar CD, Warman GR (2010) Why does insect RNA look degraded? Journal of Insect Science 10: 159.
  32. Yoshikawa K, Umetsu K, Shinzawa H et al. (1999) Determination of carbohydrate deficient transferring separated by lectin affinity chromatography for detecting chronic abuse. FEBS Letters 458: 112-116. https://doi.org/10.1016/S0014-5793(99)01137-0

Cited by

  1. Exosome isolation from hemolymph of white-spotted flower chafer, Protaetia brevitarsis (Kolbe) (Coleoptera: Scarabaeidae). vol.33, pp.2, 2016, https://doi.org/10.7852/ijie.2016.33.2.85
  2. Upregulation of Kruppel-like Factor 4 Gene expression by Allomyrina dichotoma Hemolymph in the INS-1 Pancreatic β-cells vol.26, pp.1, 2015, https://doi.org/10.15616/bsl.2020.26.1.37
  3. Exploring the Role of a Novel Peptide from Allomyrina dichotoma Larvae in Ameliorating Lipid Metabolism in Obesity vol.21, pp.22, 2015, https://doi.org/10.3390/ijms21228537
  4. Double-stranded RNA in exosomes: Potential systemic RNA interference pathway in the Colorado potato beetle, Leptinotarsa decemlineata vol.23, pp.4, 2020, https://doi.org/10.1016/j.aspen.2020.09.012
  5. RNAs on the Go: Extracellular Transfer in Insects with Promising Prospects for Pest Management vol.10, pp.3, 2021, https://doi.org/10.3390/plants10030484
  6. Morphological Characterisation of Haemocytes in the Mealworm Beetle Tenebrio molitor (Coleoptera, Tenebrionidae) vol.12, pp.5, 2015, https://doi.org/10.3390/insects12050423