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Molecular characterization of clathrin heavy chain (Chc) in Rhipicephalus haemaphysaloides and its effect on vitellogenin (Vg) expression via the clathrin-mediated endocytic pathway

Abstract

Clathrin plays an important role in arthropods, but its function in ticks has not been explored. Here, we describe the molecular characteristics of the clathrin heavy chain of the tick Rhipicephalus haemaphysaloides and its effects on yolk development. The open reading frame of the clathrin heavy chain (Chc) (Rh-Chc) gene consists of 5103 nucleotides encoding 670 amino acids, which is most closely related to that of Ixodes scapularis and relatively close to Homo sapiens and Drosophila melanogaster. Real-time qPCR revealed that Rh-Chc was expressed at all developmental stages and organs. After Rh-Chc is silenced, ticks did not feed and mortality rate was 100%. Moreover, Rh-Chc co-localized with Vitellogenin receptor (VgR) on oocyte membrane. Immunofluorescence showed that the expression of Vitellogenin (Vg) (Rh-Vg) was also closely related to Rh-Chc. Immunofluorescence showed that the expression of Vg was also closely related to Rh-Chc, Rh-Chc silencing slowed the development of oocytes in tick, and culture of ovary in vitro silenced Rh-Chc, the development of oocytes in ticks also slowed down. Overall, the results of this study indicated that Rh-Chc is a vital gene in the tick R. haemaphysaloides that plays an important role in its growth, development, and reproduction.

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References

  1. Acosta EG, Castilla V, Damonte EB (2008) Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis. J Gen Virol 89:474–484. https://doi.org/10.1099/vir.0.83357-0

  2. Avarre JC, Lubzens E, Babin PJ (2007) Apolipocrustacein, formerly vitellogenin, is the major egg yolk precursor protein in decapod crustaceans and is homologous to insect apolipophorin II/I and vertebrate apolipoprotein B. BMC Evol Biol 7:3. https://doi.org/10.1186/1471-2148-7-3

  3. Bazinet C, Katzen AL, Morgan M, Mahowald AP, Lemmon SK (1993) The Drosophila clathrin heavy chain gene: clathrin function is essential in a multicellular organism. Genetics 134:1119–1134

  4. Blariza MJ, Soria NW, Torres AG, Grosso CG, Garcia BA (2014) CDNA isolation and characterization of two vitellogenin genes in the Chagas’ disease vector Triatoma infestans (Hemiptera, Reduviidae). Gene 543:118–124. https://doi.org/10.1016/j.gene.2014.03.054

  5. Blariza MJ, Grosso CG, Garcia BA (2017) Silencing of two vitellogenin genes inhibits oviposition in the chagas disease vector Triatoma infestans (Hemiptera: Reduviidae). Am J Trop Med Hyg 97:477–480. https://doi.org/10.4269/ajtmh.16-1029

  6. Boldbaatar D, Umemiya-Shirafuji R, Liao M, Tanaka T, Xuan X, Fujisaki K (2010) Multiple vitellogenins from the Haemaphysalis longicornis tick are crucial for ovarian development. J Insect Physiol 56:1587–1598. https://doi.org/10.1016/j.jinsphys.2010.05.019

  7. Brault V, Herrbach E, Reinbold C (2007) Electron microscopy studies on luteovirid transmission by aphids. Micron 38:302–312. https://doi.org/10.1016/j.micron.2006.04.005

  8. Casartelli M, Cermenati G, Rodighiero S, Pennacchio F, Giordana B (2008) A megalin-like receptor is involved in protein endocytosis in the midgut of an insect (Bombyx mori, Lepidoptera). Am J Physiol Regul Integr Comp Physiol 295:R1290–R1300. https://doi.org/10.1152/ajpregu.00036.2008

  9. Clifton ME, Noriega FG (2011) Nutrient limitation results in juvenile hormone-mediated resorption of previtellogenic ovarian follicles in mosquitoes. J Insect Physiol 57:1274–1281. https://doi.org/10.1016/j.jinsphys.2011.06.002

  10. Cong L, Yang WJ, Jiang XZ, Niu JZ, Shen GM, Ran C, Wang JJ (2015) The essential role of vitellogenin receptor in ovary development and vitellogenin uptake in Bactrocera dorsalis (Hendel). Int J Mol Sci 16:18368–18383. https://doi.org/10.3390/ijms160818368

  11. Grant B, Hirsh D (1999) Receptor-mediated endocytosis in the Caenorhabditis elegans oocyte. Mol Biol Cell 10:4311–4326. https://doi.org/10.1091/mbc.10.12.4311

  12. Hansen IA, Attardo GM, Park JH, Peng Q, Raikhel AS (2004) Target of rapamycin-mediated amino acid signaling in mosquito anautogeny. Proc Natl Acad Sci USA 101:10626–10631. https://doi.org/10.1073/pnas.0403460101

  13. Horigane M, Shinoda T, Honda H, Taylor D (2010) Characterization of a vitellogenin gene reveals two phase regulation of vitellogenesis by engorgement and mating in the soft tick Ornithodoros moubata (Acari: Argasidae). Insect Mol Biol 19:501–515. https://doi.org/10.1111/j.1365-2583.2010.01007.x

  14. Inoue T, Hayashi T, Takechi K, Agata K (2007) Clathrin-mediated endocytic signals are required for the regeneration of, as well as homeostasis in, the planarian CNS. Development 134:1679–1689. https://doi.org/10.1242/dev.02835

  15. Jatuyosporn T, Supungul P, Tassanakajon A, Krusong K (2014) The essential role of clathrin-mediated endocytosis in yellow head virus propagation in the black tiger shrimp Penaeus monodon. Dev Comp Immunol 44:100–110. https://doi.org/10.1016/j.dci.2013.11.017

  16. Kokoza VA, Snigirevskaya ES, Raikhel AS (1997) Mosquito clathrin heavy chain: analysis of protein structure and developmental expression in the ovary during vitellogenesis. Insect Mol Biol 6:357–368

  17. Li A, Sadasivam M, Ding JL (2003) Receptor–ligand interaction between vitellogenin receptor (VtgR) and vitellogenin (Vtg), implications on low density lipoprotein receptor and apolipoprotein B/E. The first three ligand-binding repeats of VtgR interact with the amino-terminal region of Vtg. J Biol Chem 278:2799–2806. https://doi.org/10.1074/jbc.m205067200

  18. Lu K, Chen X, Liu WT, Zhang XY, Chen MX, Zhou Q (2016) Nutritional signaling regulates vitellogenin synthesis and egg development through juvenile hormone in Nilaparvata lugens (Stal). Int J Mol Sci 17:269. https://doi.org/10.3390/ijms17030269

  19. McMahon HT, Boucrot E (2011) Molecular mechanism and physiological functions of clathrin-mediated endocytosis. Nat Rev Mol Cell Biol 12:517–533. https://doi.org/10.1038/nrm3151

  20. Mhashilkar AS, Adapa SR, Jiang RH, Williams SA, Zaky W, Slatko BE, Luck AN, Moorhead AR, Unnasch TR (2016) Phenotypic and molecular analysis of the effect of 20-hydroxyecdysone on the human filarial parasite Brugia malayi. Int J Parasitol 46:333–341. https://doi.org/10.1016/j.ijpara.2016.01.005

  21. Mizuta H, Mushirobira Y, Nagata J, Todo T, Hara A, Reading BJ, Sullivan CV, Hiramatsu N (2017) Ovarian expression and localization of clathrin (Cltc) components in cutthroat trout, Oncorhynchus clarkii: evidence for Cltc involvement in endocytosis of vitellogenin during oocyte growth. Comp Biochem Physiol A 212:24–34. https://doi.org/10.1016/j.cbpa.2017.06.021

  22. O’Halloran TJ, Anderson RG (1992) Clathrin heavy chain is required for pinocytosis, the presence of large vacuoles, and development in Dictyostelium. J Cell Biol 118:1371–1377

  23. Payne GS, Schekman R (1985) A test of clathrin function in protein secretion and cell growth. Science 230:1009–1014

  24. Pearse BM (1975) Coated vesicles from pig brain: purification and biochemical characterization. J Mol Biol 97:93–98

  25. Qiu J, He Y, Zhang J, Kang K, Li T, Zhang W (2016) Discovery and functional identification of fecundity-related genes in the brown planthopper by large-scale RNA interference. Insect Mol Biol 25:724–733. https://doi.org/10.1111/imb.12257

  26. Raikhel AS, Dhadialla TS (1992) Accumulation of yolk proteins in insect oocytes. Annu Rev Entomol 37:217–251. https://doi.org/10.1146/annurev.en.37.010192.001245

  27. Ruscetti T, Cardelli JA, Niswonger ML, O’Halloran TJ (1994) Clathrin heavy chain functions in sorting and secretion of lysosomal enzymes in Dictyostelium discoideum. J Cell Biol 126:343–352

  28. Sappington TW, Raikhel AS (1998) Molecular characteristics of insect vitellogenins and vitellogenin receptors. Insect Biochem Mol Biol 28:277–300

  29. Singh NK, Pakkianathan BC, Kumar M, Prasad T, Kannan M, Konig S, Krishnan M (2013) Vitellogenin from the silkworm, Bombyx mori: an effective anti-bacterial agent. PLoS ONE 8:e73005. https://doi.org/10.1371/journal.pone.0073005

  30. Snigirevskaya ES, Sappington TW, Raikhel AS (1997) Internalization and recycling of vitellogenin receptor in the mosquito oocyte. Cell Tissue Res 290:175–183

  31. Veerana M, Kubera A, Ngernsiri L (2014) Analysis of the vitellogenin gene of rice moth, Corcyra cephalonica Stainton. Arch Insect Biochem Physiol 87:126–147. https://doi.org/10.1002/arch.21185

  32. Wu K, Hoy MA (2014) Clathrin heavy chain is important for viability, oviposition, embryogenesis and possibly, systemic RNAi response in the predatory mite Metaseiulus occidentalis. PLoS ONE 9:e110874. https://doi.org/10.1371/journal.pone.0110874

  33. Ybe JA (2014) Novel clathrin activity: developments in health and disease. Biomol Concepts 5:175–182. https://doi.org/10.1515/bmc-2013-0040

  34. Zhong R, Ding TB, Niu JZ, Xia WK, Liao CY, Dou W, Wang JJ (2015) Molecular characterization of vitellogenin and its receptor genes from citrus red mite, Panonychus citri (McGregor). Int J Mol Sci 16:4759–4773. https://doi.org/10.3390/ijms16034759

  35. Zhou X, Fabian L, Bayraktar JL, Ding HM, Brill JA, Chang HC (2011) Auxilin is required for formation of golgi-derived clathrin-coated vesicles during Drosophila spermatogenesis. Development 138:1111–1120. https://doi.org/10.1242/dev.057422

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Acknowledgements

This work was supported by a Grant (2017YFD0501200) from the National Key Research and Development Program of China. The protocols were approved by the Institutional Animal Care and Use Committee of the Shanghai Veterinary Research Institute (IACUC Approval Number shvri-ra-2018020098) and authorized by the Animal Ethical Committee of Shanghai Veterinary Research Institute.

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Correspondence to Jinlin Zhou.

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Kuang, C., Wang, F., Zhou, Y. et al. Molecular characterization of clathrin heavy chain (Chc) in Rhipicephalus haemaphysaloides and its effect on vitellogenin (Vg) expression via the clathrin-mediated endocytic pathway. Exp Appl Acarol 80, 71–89 (2020). https://doi.org/10.1007/s10493-019-00438-5

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Keywords

  • Clathrin-mediated endocytosis
  • Clathrin heavy chain
  • Vitellogenin
  • Vitellogenin Receptor