Eur. J. Entomol. 114: 235-248, 2017 | DOI: 10.14411/eje.2017.029

Juvenile hormone analogue, fenoxycarb, modulates ecdysone-triggered transcriptional hierarchy during programmed cell death of midgut in silkworm, Bombyx mori (Lepidoptera: Bombycidae)

Ebru GONCU, Ramazan URANLI, Osman PARLAK
Ege University, Faculty of Science, Department of Biology, 35100 Bornova - Izmir, Turkey; e-mails: ebru.goncu@ege.edu.tr, uranli.ramazan@gmail.com, osman.parlak@ege.edu.tr

Steroid hormone 20-hydroxyecdysone and the sesquiterpenoid juvenile hormone are the main regulators of insect development; however, it is unclear how they interact in the regulation of metamorphic events. Using the silkworm, Bombyx mori, we show that the juvenile hormone analogue fenoxycarb affects the cascade of ecdysone regulated genes that control the programmed cell death in the larval midgut. Morphological changes that occur during cell death were investigated by studying cross-sections of the midgut stained with hematoxylin and eosin. Apoptosis-specific DNA fragmentation was detected using TUNEL assay. Expression patterns of genes ATG8 and ATG12, which were used as indicators of autophagy, and genes of the ecdysone-regulated gene cascade were examined using real-time quantitative polymerase chain reaction. Fenoxycarb application on day 0 of the 5th larval instar extended the feeding period and postponed programmed cell death in mature larval midgut. This effect was probably due to a delay in ecdysone secretion and associated changes in gene expression were mostly not a direct response to the fenoxycarb. However, differences in the gene expression patterns in the control and fenoxycarb treated insects during the prepupal and early pupal stages indicated that fenoxycarb may also exert a more direct effect on some genes of the ecdysone regulated gene cascade.

Keywords: Lepidoptera, Bombycidae, apoptosis, autophagy, Bombyx mori, ecdysone related genes, juvenile hormone, midgut

Received: November 28, 2016; Accepted: March 27, 2017; Published online: May 5, 2017Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
GONCU, E., URANLI, R., & PARLAK, O. (2017). Juvenile hormone analogue, fenoxycarb, modulates ecdysone-triggered transcriptional hierarchy during programmed cell death of midgut in silkworm, Bombyx mori (Lepidoptera: Bombycidae). Eur. J. Entomol.114(1), 2017.000. doi: 10.14411/eje.2017.029.
Download citation

References

  1. Baehrecke E.H. 2000: Steroid regulation of programmed cell death during Drosophila development. - Cell Death Different. 7: 1057-1062. Go to original source...
  2. Bergmeyer H.U., Gawehn K. & Grassl M. 1974: Enzymes as biochemical reagents, acid phosphatase. In Bergmeyer H.U. (ed.): Methods of Enzymatic Analysis. Vol. 1. 2nd ed. Academic Press, New York, pp. 495-496.
  3. Bernardo T.J. & Dubrovsky E.B. 2012: The Drosophila juvenile hormone receptor candidates methoprene-tolerant (MET) and germ cell-expressed (GCE) utilize a conserved LIXXL motif to bind the FTZ-F1 nuclear receptor. - J. Biol. Chem. 2: 7821-7833. Go to original source...
  4. Denton D., Shravage B., Simin R., Mills K., Berry D.L., Baehrecke E.H. & Kumar S. 2009: Autophagy, not apoptosis, is essential for midgut cell death in Drosophila. - Curr. Biol. 19: 1741-1746. Go to original source...
  5. Denton D., Shravage B., Simin R., Baehrecke E.H. & Kumar S. 2010: Larval midgut destruction in Drosophila: Not dependent on caspases but suppressed by the loss of autophagy. - Autophagy 6: 163-165. Go to original source...
  6. Dubrovsky E.B., Dubrovskaya V.A. & Berger E.M. 2004: Hormonal regulation and functional role of Drosophila E75A orphan nuclear receptor in the juvenile hormone signaling pathway. - Dev. Biol. 268: 258-270. Go to original source...
  7. Dubrovsky E.B., Dubrovskaya V.A., Bernardo T., Otte V., Difilippo R. & Bryan H. 2011: The Drosophila FTZ-F1 nuclear receptor mediates juvenile hormone activation of E75A gene expression through an intracellular pathway. - J. Biol. Chem. 286: 33689-33700. Go to original source...
  8. Franzetti E., Huang Z.J., Shi Y.X, Xie K., Deng X.J., Li J.P., Li Q.R., Yang W.Y., Zeng W.N., Casartelli M., Deng H.M., Cappellozza S., Grimaldi A., Xia Q., Feng Q., Cao Y. & Tettamanti G. 2012: Autophagy precedes apoptosis during the remodeling of silkworm larval midgut. - Apoptosis 17: 305- 324. Go to original source...
  9. Franzetti E., Romanelli D., Caccia S., Cappellozza S., Congiu T., Rajagopalan M., Grimaldi A., de Eguileor M., Casartelli M. & Tettamanti G. 2015: The midgut of the silkmoth Bombyx mori is able to recycle molecules derived from degeneration of the larval midgut epithelium. - Cell Tissue Res. 361: 509-528. Go to original source...
  10. Gibson U.E., Heid C.A. & Williams P.M. 1996: A novel method for real time quantitative RT-PCR. - Genome Res. 6: 995-1001. Go to original source...
  11. Goncu E. & Parlak O. 2012: The effect of the juvenile hormone analog, fenoxycarb, on ecdysone receptor B1 expression in the midgut of Bombyx mori during larval-pupal metamorphosis. - Folia Histochem. Cytobiol. 50: 52-57. Go to original source...
  12. Goncu E. & Parlak O. 2011: The influence of juvenile hormone analogue, fenoxycarb on the midgut remodeling in Bombyx mori (L., 1758) (Lepidoptera: Bombycidae) during larval-pupal metamorphosis. - Turk. J. Entomol. 35: 179-194.
  13. Goncu E., Uranli R., Selek G. & Parlak O. 2016: Developmental expression of ecdysone-related genes associated with metamorphic changes during midgut remodeling of silkworm Bombyx mori (Lepidoptera: Bombycidae). - J. Insect Sci. 16: 86, 13 pp. Go to original source...
  14. Hakim R.S., Caccia S., Loeb M. & Smagghe G. 2009: Primary culture of insect midgut cells. - In Vitro Cell. Dev. Biol. Anim. 45: 106-110.
  15. Hiruma K., Shinoda T., Malone F. & Riddiford L.M. 1999: Juvenile hormone modulates 20-hydroxyecdysone-inducible ecdysone receptor and ultraspiracle gene expression in the tobacco hornworm, Manduca sexta. - Dev. Genes Evol. 209: 18-30. Go to original source...
  16. Huang J.H., Lozano J. & Belles X. 2013: Broad-complex functions in postembryonic development of the cockroach Blattella germanica shed new light on the evolution of insect metamorphosis. - Biochim. Biophys. Acta 1830: 2178-2187. Go to original source...
  17. Jiang C., Baehreck E.H. & Thummel C.S. 1997: Steroid regulated programmed cell death during Drosophila metamorphosis. - Development 22: 4673-4683.
  18. Jiang C., Lamblin A.F., Steller H. & Thummel C.S. 2000: A steroid-triggered transcriptional hierarchy controls salivary gland cell death during Drosophila metamorphosis. - Mol. Cell 5: 445-455. Go to original source...
  19. Jones G. & Sharp P.A. 1997: Ultraspiracle: An invertebrate nuclear receptor for juvenile hormones. - Proc. Natl. Acad. Sci. USA 94: 13499-13503. Go to original source...
  20. Kamimura M. & Kiuchi M. 1998: Effects of a juvenile hormone analog, fenoxycarb, on 5th stadium of the larvae of the silkworm, Bombyx mori (Lepidoptera: Bombycidae). - Appl. Entomol. Zool. 33: 333-338. Go to original source...
  21. King-Jones K. & Thummel C.S. 2005: Nuclear receptors - a perspective from Drosophila. - Nature Rev. Genet. 6: 311-323. Go to original source...
  22. Khoa D.B. & Takeda M. 2012: Expression of autophagy 8 (Atg8) and its role in the midgut and other organs of the greater wax moth, Galleria mellonella, during metamorphic remodeling and under starvation. - Insect Mol. Biol. 21: 473-487. Go to original source...
  23. Konopova B., Smykal V. & Jindra M. 2011: Common and distinct roles of juvenile hormone signaling genes in metamorphosis of holometabolous and hemimetabolous insects. - PLoS One 6(12): e28728, 7 pp. Go to original source...
  24. Lan Q., Hiruma K., Hu X., Jindra M. & Riddiford L.M. 1999: Activation of a delayed-early gene encoding MHR3 by the ecdysone receptor heterodimer EcR-B1-USP-1 but not by EcR-B1-USP-2. - Mol. Cell. Biol. 19: 4897-4906. Go to original source...
  25. Lee C.Y., Simon C.R., Woodard C.T. & Baehrecke E.H. 2002: Genetic mechanism for the stage-and tissue-specific regulation of steroid triggered programmed cell death in Drosophila. - Devel. Biol. 252: 138-148. Go to original source...
  26. Lockshin R.A. & Williams C.M. 1965: Programmed cell death - I. Cytology of degeneration in the intersegmental muscles of the silkmoth. - J. Insect Physiol. 11: 123-133. Go to original source...
  27. Matsumoto H., Ueno C., Nakamura Y., Kinjoh T., Ito Y., Shimura S., Noda H., Imanishi S., Mita K., Fujiwara H., Hiruma K., Shinoda T. & Kamimura M. 2015: Identification of two juvenile hormone inducible transcription factors from the silkworm, Bombyx mori. - J. Insect Physiol. 80: 31-41. Go to original source...
  28. Nishiura J.T. 2002: Coordinated morphological changes in midgut, imaginal discs, and respiratory trumpets during metamorphosis of Aedes aegypti (Diptera: Culicidae). - Ann. Entomol. Soc. Am. 95: 498-505. Go to original source...
  29. Nishiura J.T., Ray K. & Murray J. 2005: Expression of nuclear receptor - transcription factor genes during Aedes aegypti midgut metamorphosis and the effect of methoprene on expression. - Insect Biochem. Mol. Biol. 35: 561-573. Go to original source...
  30. Parthasarathy R. & Palli S.R. 2007: Stage- and cell-specific expression of ecdysone receptors and ecdysone-induced transcription factors during midgut remodeling in the yellow fever mosquito, Aedes aegypti. - J. Insect Physiol. 53: 216-229. Go to original source...
  31. Parthasarathy R. & Palli S.R. 2009: Molecular analysis of juvenile hormone analog action in controlling the metamorphosis of the red flour beetle, Tribolium castaneum. - Arch. Insect Biochem. Physiol. 70: 57-70. Go to original source...
  32. Restifo L.L. & Wilson T.G. 1998: A juvenile hormone agonist reveals distinct developmental pathways mediated by ecdysone-inducible Broad complex transcription factors. - Dev. Genet. 22: 141-159. Go to original source...
  33. Riddiford L.M. 2008: Juvenile hormone action: A 2007 perspective. - J. Insect Physiol. 54: 895-901. Go to original source...
  34. Santhanam A., Peng W.H., Yu Y.T., Sang T.K., Chen G.C. & Meng T.C. 2014: Ecdysone-induced receptor tyrosine phosphatase PTP52F regulates Drosophila midgut histolysis by enhancement of autophagy and apoptosis. - Mol. Cell. Biol. 34: 1594-1606. Go to original source...
  35. Siaussat D., Bozzolan F., Queguiner I., Porcheron P. & Debernard S. 2004: Effects of juvenile hormone on 20-hydroxyecdysone-inducible EcR, HR3, E75 gene expression in imaginal wing cells of Plodia interpunctella (Lepidoptera). - Eur. J. Biochem. 271: 3017-3027. Go to original source...
  36. Smagghe G., Vanhassel W., Moeremans C., Wilde D., Goto S., Loeb M.J., Blacburn M.B. & Hakim R.S. 2005: Stimulation of midgut stem cell proliferation and differentiation by insect hormones and peptides. - Ann. N. Y. Acad. Sci. 1040: 472-475. Go to original source...
  37. Suzuki Y., Truman J.W. & Riddiford L.M. 2008: The role of Broad in the development of Tribolium castaneum: Implications for the evolution of the holometabolous insect pupa. - Development 135: 569-577. Go to original source...
  38. Talbot W.S., Swyryd E.A. & Hogness D.S. 1993: Drosophila tissues with different metamorphic responses to ecdysone express different ecdysone receptor isoforms. - Cell 73: 1323-1337. Go to original source...
  39. Tereshima J., Yasuhara N., Iwami M. & Sakurai S. 2000: Programmed cell death triggered by insect steroid hormone, 20-hydroxyecdysone in the anterior silk gland of the silkworm, Bombyx mori. - Dev. Genes Evol. 210: 545-558. Go to original source...
  40. Tettamanti G., Grimaldi A., Casartelli M., Ambrosetti E., Ponti B., Congiu T., Ferrarese F., Rivas-Pena M.R., Pennacchio F. & De Eguileor M. 2007: Programmed cell death and stem cell differentiation are responsible for midgut replacement in Heliothis virescens during prepupal instar. - Cell Tissue Res. 330: 345-359. Go to original source...
  41. Wigglesworth V.B. 1963: The action of moulting hormone and juvenile hormone at the cellular level in Rhodnius prolixus. - J. Exp. Biol. 40: 231-245.
  42. Wu Y., Parthasarathy R., Bai H. & Pallii S.R. 2006: Mechanisms of midgut remodeling: Juvenile hormone analog methoprene blocks midgut metamorphosis by modulating ecdysone action. - Mechan. Dev. 123: 530-547. Go to original source...
  43. Zhou B., Hiruma K., Jindra M., Shinoda T., Segraves W.A., Malone F. & Riddiford L.M. 1998: Regulation of the transcription factor E75 by 20-hydroxyecdysone and juvenile hormone in the epidermis of the tobacco hornworm, Manduca sexta, during larval molting and metamorphosis. - Dev. Biol. 193: 127-138. Go to original source...