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Apoptosis

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Ultraspiracle-independent anti-apoptotic function of ecdysone receptors is required for the survival of larval peptidergic neurons via suppression of grim expression in Drosophila melanogaster

  • Gyunghee Lee
  • Ritika Sehgal
  • Zixing Wang
  • Jae H. Park
Article
  • 1 Downloads

Abstract

In Drosophila melanogaster a significant number of heterogenous larval neurons in the central nervous system undergo metamorphosis-associated programmed cell death, termed metamorphoptosis. Interestingly distinct groups of doomed larval neurons are eliminated at different metamorphic phases. Although ecdysone hormonal signaling via nuclear ecdysone receptors (EcRs) is known to orchestrate the neuronal metamorphoptosis, little is known about how this signaling controls such diverse neuronal responses. Crustacean cardioactive peptide (CCAP)-producing neurons in the ventral nerve cord are developmentally programmed to die shortly after adult emergence. In this study, we show that disruption of endogenous EcR function by ectopic expression of dominant negative forms of EcRs (EcRDN) causes premature death of larval CCAP neurons in a caspase-dependent manner. This event is rescued by co-expression of individual EcR isoforms. Furthermore, larval CCAP neurons are largely normal in ecr mutants lacking either EcR-A or EcR-B isoforms, suggesting that EcR isoforms redundantly function to protect larval CCAP neurons. Of surprise, a role of Ultraspiracle (Usp), a canonical partner of EcR, is dispensable in the protection of CCAP neurons, whereas both EcR and Usp are required for inducing metamorphoptosis of vCrz neurons shortly after prepupal formation. As a downstream, grim is an essential cell death gene for the EcRDN-mediated CCAP neuronal death, while either hid or rpr function is dispensable. Together, our results suggest that Usp-independent EcR actions protect CCAP neurons from their premature death by repressing grim expression until their normally scheduled apoptosis at post-emergence. Our studies highlight two opposite roles played by EcR function for metamorphoptosis of two different peptidergic neuronal groups, proapoptotic (vCrz) versus antiapoptotic (CCAP), and propose that distinct death timings of doomed larval neurons are determined by differential signaling mechanisms involving EcR.

Keywords

Metamorphoptosis Ultraspiracle Ecdysone receptor Central nervous system Peptidergic neurons Grim Apoptosis 

Abbreviations

AE

After eclosion

APF

After puparium formation

CNS

Central nervous system

PCD

Programmed cell death

VNC

Ventral nerve cord

CCAP

Crustacean cardioactive peptide

Crz

Corazonin

EcR

Ecdysone receptor

USP

Ultraspiracle

Notes

Acknowledgements

We want to express our gratitude to many people for their kind donation of various research materials; B. White (NIH) for the anti-bursicon, K. White (Mass General Hospital) for hid mutants, T. Lee (Janelia Farm) for UAS-miusp line, M. Bender (Univ. of Georgia) for ecr mutants, S. Robinow (Univ. of Hawaii) for UAS-EcR, P. Cherbas (Indiana Univ.) for UAS-EcRDN lines, B. Hay (Caltech) for UAS-miRGH and UAS-migrim lines, T. Lee (Janelia farm) for usp3 MARCM lines. This work was supported by an NIH Grant (R15-GM114741) and by Hunsicker research incentive Grant (Univ. of Tennessee).

Supplementary material

10495_2019_1514_MOESM1_ESM.pdf (2.7 mb)
Supplemental Fig. 1 Developmental phenotypes by A9-gal4 driven expression of EcRW650A or Usp3. a–eEcRW650A expression caused various developmental defects in larval stages (a–b) and in pupal stages (c–e), whereas Usp3 expression did not affect larval growth but led to death of pharate adults inside the pupal case (f) (PDF 2791 KB)

References

  1. 1.
    Ishizuya-Oka A, Hasebe T, Shi Y-B (2010) Apoptosis in amphibian organs during metamorphosis. Apoptosis 15:350–364CrossRefGoogle Scholar
  2. 2.
    Yin VP, Thummel CS (2005) Mechanisms of steroid-triggered programmed cell death in Drosophila. Sem Cell Dev Biol 16:237–243CrossRefGoogle Scholar
  3. 3.
    Lee G, Kim J, Kim Y, Yoo S, Park JH (2018) Identifying and monitoring neurons that undergo metamorphosis-regulated cell death (metamorphoptosis) by a neuron-specific caspase sensor (Casor) in Drosophila melanogaster. Apoptosis 23:41–53CrossRefGoogle Scholar
  4. 4.
    Truman JW, Taylor BJ, Awad TA (1993) Formation of the adult nervous system. In: Bate M, Arias AM (eds) Development of Drosophila melanogaster. Cold Spring Harbor Press, New York, pp 1245–1275Google Scholar
  5. 5.
    Truman JW (1996) Metamorphosis of the insect nervous system. In: Gilbert LI, Tata JR, Atkinson BG (ed) Metamorphosis: postembryonic reprogramming of gene expression in Amphibian and insect cells. Academic Press, Cambridge, pp. 283–320CrossRefGoogle Scholar
  6. 6.
    Weeks JC (2003) Thinking globally, acting locally: steroid hormone regulation of the dendritic architecture, synaptic connectivity and death of an individual neuron. Prog Neurobiol 70:421–442CrossRefGoogle Scholar
  7. 7.
    Yaniv SP, Schuldiner O (2016) A fly’s view of neuronal remodeling. WIREs Dev Biol 5:618–635CrossRefGoogle Scholar
  8. 8.
    Pinto-Teixeira F, Konstantinides N, Desplan C (2016) Programmed cell death acts at different stages of Drosophila neurodevelopment to shape te central nervous system. FEBS Lett 590:2435–2453CrossRefGoogle Scholar
  9. 9.
    Choi Y-J, Lee G, Park JH (2006) Programmed cell death mechanisms of identifiable peptidergic neurons in Drosophila melanogaster. Development 133:2223–2232CrossRefGoogle Scholar
  10. 10.
    Lee G, Wang Z, Sehgal R, Kikuno K, Chen C-H, Hay B, Park JH (2011) Drosophila caspases involved in developmentally regulated programmed cell death of peptidergic neurons during early metamorphosis. J Comp Neurol 519:34–48CrossRefGoogle Scholar
  11. 11.
    Lee G, Kikuno K, Sehgal R, Wang Z, Nair S, Chen C-H, Hay B, Park JH (2013) Essential role of grim-led programmed cell death for the establishment of Corazonin-producing peptidergic nervous system during embryogenesis and metamorphosis in Drosophila melanogaster. Biol Open 2:283–294CrossRefGoogle Scholar
  12. 12.
    Winbush A, Weeks JC (2011) Steroid-triggered, cell-autonomous death of a Drosophila motoneurons during metamorphosis. Neural Dev 6:15CrossRefGoogle Scholar
  13. 13.
    Robinow S, Talbot WS, Hogness DS, Truman JW (1993) Programmed cell death in the Drosophila CNS is ecdysone-regulated and coupled with a specific ecdysone receptor isoform. Development 119:1251–1259Google Scholar
  14. 14.
    Draizen TA, Ewer J, Robinow S (1999) Genetic and hormonal regulation of the death of peptidergic neurons in the Drosophila central nervous system. J Neurobiol 38:455–465CrossRefGoogle Scholar
  15. 15.
    Lee G, Kikuno K, Nair S, Park JH (2013) Mechanisms of postecdysis-associated programmed cell death of peptidergic neurons in Drosophila melanogaster. J Comp Neurol 521:3972–3991Google Scholar
  16. 16.
    Dai JD, Gilbert LI (1991) Metamorphosis of the corpus allatum and degeneration of the prothoracic glands during the larval–pupal–adult transformation of Drosophila melanogaster: a cytophysiological analysis of the ring gland. Dev Biol 144:309–326CrossRefGoogle Scholar
  17. 17.
    Robinow S, Draizen TA, Truman JW (1997) Genes that induce apoptosis: transcriptional regulation in identified, doomed neurons of the Drosophila CNS. Dev Biol 190:206–213CrossRefGoogle Scholar
  18. 18.
    Park JH, Shroeder AJ, Helfrich-Förster C, Jackson FR, Ewer J (2003) Targeted ablation of CCAP neuropeptide-containing neurons of Drosophila causes specific defects in execution and circadian timing of ecdysis behavior. Development 130:2645–2656CrossRefGoogle Scholar
  19. 19.
    Luan H, Lemon WC, Peabody NC, Pohl JB, Zelensky PK, Wang D, Nitabach MN, Holmes TC, White BH (2006) Functional dissection of a neuronal network required for cuticle tanning and wing expansion in Drosophila. J Neurosci 26:573–584CrossRefGoogle Scholar
  20. 20.
    Choi S-H, Lee G, Monahan P, Park JH (2008) Spatial regulation of Corazonin neuropeptide expression requires multiple cis-acting elements in Drosophila melanogaster. J Comp Neurol 507:1184–1195CrossRefGoogle Scholar
  21. 21.
    Schubiger M, Wade A, Carney GE, Truman JW, Bender M (1998) Drosophila EcR-B ecdysone receptor isoforms are required for larval molting and for neuron remodeling during metamorphosis. Development 125:2053–2062Google Scholar
  22. 22.
    Bender M, Imam FB, Talbot WS, Ganetzky B, Hogness DS (1997) Drosophila ecdysone receptor mutations reveal functional differences among receptor isoforms. Cell 91:777–788CrossRefGoogle Scholar
  23. 23.
    Carney GE, Robertson A, Davis MB, Bender M (2004) Creation of EcR isoform-specific mutations in Drosophila melanogaster via local P element transposition, imprecise P element excision, and male recombination. Mol Genet Genomics 271:282–290CrossRefGoogle Scholar
  24. 24.
    Grether ME, Abrams JM, Agapite J, White K, Steller K (1995) The head involution defective gene of Drosophila melanogaster functions in programmed cell death. Genes Dev 9:1694–1708CrossRefGoogle Scholar
  25. 25.
    Moon N-S, Stefano LD, Morris EJ, Patel R, White K, Dyson NJ (2008) E2F and p53 induce apoptosis independently during Drosophila development but intersect in the context of DNA damage. PLoS Genet 4(8), e1000153Google Scholar
  26. 26.
    Chen C-H, Huang H, Ward CM, Su JT, Schaffer LV, Guo M, Hay BA (2007) A synthetic maternal-effect selfish genetic element drives population replacement in Drosophila. Science 316:597–600CrossRefGoogle Scholar
  27. 27.
    Lin S, Huang Y, Lee T (2009) Nuclear receptor unfulfilled regulates axonal guidance and cell identity of Drosophila mushroom body neurons. PLoS ONE 4(12):e8392CrossRefGoogle Scholar
  28. 28.
    Cherbas L, Hu X, Zhimulev I, Belyaeva E, Cherbas P (2003) EcR isoforms in Drosophila: testing tissue-specific requirements by targeted blockade and rescue. Development 130:271–284CrossRefGoogle Scholar
  29. 29.
    Lee T, Marticke S, Sung C, Robinow S, Luo L (2000) Cell-autonomous requirement of the USP/EcR-B ecdysone receptor for mushroom body neuronal remodeling in Drosophila. Neuron 28:807–818CrossRefGoogle Scholar
  30. 30.
    Henrich VC, Szekely AA, Kim SJ, Brown NE, Antoniewski C, Hayden MA, Lepesant J-A, Gilbert LI (1994) Expression and function of the ultraspiracle (usp) gene during development of Drosophila melanogaster. Dev Biol 165:38–52CrossRefGoogle Scholar
  31. 31.
    Talbot WS, Swyryd EA, Hogness DS (1993) Drosophila tissues with different metamorphic responses to ecdysone express different ecdysone receptor isoforms. Cell 73:1323–1337CrossRefGoogle Scholar
  32. 32.
    Diao F, Mena W, Shi J, Park D, Diao F, Taghert P, Ewer J, White BH (2016) The splice isoforms of the Drosophila ecdysis triggering hormone receptor have developmentally distinct roles. Genetics 202:175–189CrossRefGoogle Scholar
  33. 33.
    Yoo S, Lam H, Lee C, Lee G, Park JH (2017) Cloning and functional characterizations of an apoptogenic Hid gene in the Scuttle Fly, Megaselia scalaris (Diptera; Phoridae). Gene 604:9–21CrossRefGoogle Scholar
  34. 34.
    Truman JW, Talbot WS, Fahrbach SE, Hogness DS (1994) Ecdysone receptor expression in the CNS correlates with stage-specific responses to ecdysteroids during Drosophila and Manduca development. Development 120:219–234Google Scholar
  35. 35.
    Zhao T, Gu T, Rice HC, McAdams KL, Roark KM, Lawson K, Gauthier SA, Reagan KL, Hewes RS (2008) A Drosophila gain-of-function screen for candidate genes involved in steroid-dependent neuroendocrine cell remodeling. Genetics 178:883–901CrossRefGoogle Scholar
  36. 36.
    Yao T, Segraves WA, Oro AE, McKeown M, Evans RM (1992) Drosophila ultraspiracle modulates ecdysone receptor function via heterodimer formation. Cell 71:63–72CrossRefGoogle Scholar
  37. 37.
    Thomas HE, Stunnenberg HG, Stewart AF (1993) Heterodimerization of the Drosophila ecdysone receptor with retinoid X receptor and ultraspiracle. Nature 362:471–475CrossRefGoogle Scholar
  38. 38.
    Oro AE, McKeown M, Evans RM (1992) The Drosophila retinoid X receptor homolog ultraspiracle functions in both female reproduction and eye morphogenesis. Development 115:449–462Google Scholar
  39. 39.
    Lee G, Park JH (2004) Hemolymph sugar homeostasis and starvation-induced hyperactivity affected by genetic manipulations of the adipokinetic hormone-encoding gene in Drosophila melanogaster. Genetics 167:311–323CrossRefGoogle Scholar
  40. 40.
    Haerry TE, Khalsa O, O’connor MB, Wharton KA (1998) Synergistic signaling by two BMP ligands through the SAX and TKV receptors controls wing growth and patterning in Drosophila. Development 125:3977–3987Google Scholar
  41. 41.
    Goyal L, McCall K, Agapite J, Hartwieg E, Steller H (2000) Induction of apoptosis by Drosophila reaper, hid, and grim through inhibition of IAP function. EMBO J 19:589–597CrossRefGoogle Scholar
  42. 42.
    Dewey EM, McNabb SL, Ewer J, Kuo GR, Takanishi CL, Truman JW, Honegger H-W (2004) Identification of the gene encoding bursicon, an insect neuropeptide responsible for cuticle sclerotization and wing spreading. Curr Biol 14:1208–1213CrossRefGoogle Scholar
  43. 43.
    Luo C-W, Dewey EM, Sudo S, Ewer J, Hsu SY, Honegger H-W, Hsueh AJW (2005) Bursicon, the insect cuticle-hardening hormone, is a heterodimeric cystine knot protein that activates G protein-coupled receptor LGR2. Proc Natl Acad Sci USA 102:2820–2825CrossRefGoogle Scholar
  44. 44.
    Kim YJ, Zitnan D, Galizia CG, Cho KH, Adams ME (2006) A command chemical triggers an innate behavior by sequential activation of multiple peptidergic ensembles. Curr Biol 16:1395–1407CrossRefGoogle Scholar
  45. 45.
    Handler AM (1982) Ecdysteroid titers during pupal and adult development in Drosophila melanogaster. Dev Biol 93:73–82CrossRefGoogle Scholar
  46. 46.
    Sutherland J, Kozlova T, Tzertzinis G, Kafatos F (1995) Drosophila hormone receptor 38: a second partner for Drosophila USP suggests an unexpected role for nuclear receptors of the nerve growth factor-induced protein B type. Proc Natl Acad Sci USA 92:7966–7970CrossRefGoogle Scholar
  47. 47.
    Buszczak M, Segraves WA (1998) Drosophila metamorphosis: the only way is USP? Curr Biol 8:879–882CrossRefGoogle Scholar
  48. 48.
    Siddiqui AN, Siddiqui N, Khan RA, Kalam A, Jabir NR, Kamal MA, Firoz CK, Tabrez S (2016) Neuroprotective role of steroidal sex hormones: an overview. CNS Neurosci Ther 22:342–350CrossRefGoogle Scholar
  49. 49.
    Sasaya H, Yasuzumi K, Maruoka H, Fujita A, Kato Y, Waki T, Shimoke K, Ikeuchi T (2012) Apoptosis-inducing activity of endocrine-disrupting chemicals in cultured PC12 cells. Adv Biol Chem 2:92–105CrossRefGoogle Scholar
  50. 50.
    Oka T, Adati N, Shinkai T, Sakuma K, Nishmura T, Kurose K (2003) Bisphenol A induces apoptosis in central neural cells during early development of Xenopus laevis. Biochem Biophys Res Commun 312:877–882CrossRefGoogle Scholar
  51. 51.
    Masuo Y, Ishido M (2011) Neurotoxicity of endocrine disruptors: possible involvement in brain development and neurodegeneration. J Toxcol Environ Health part B 14:346–369CrossRefGoogle Scholar
  52. 52.
    Nunez JL, Lauschke DM, Juraska JM (2001) Cell death in the development of the posterior cortex in male and female rats. J Comp Neurol 436:32–41CrossRefGoogle Scholar
  53. 53.
    Nunez JL, Sodhi J, Juraska JM (2002) Ovarian hormones after postnatal day 20 reduce neuron number in the rat primary visual cortex. J Neurobiol 52:312–321CrossRefGoogle Scholar
  54. 54.
    Tsukahara S (2009) Sex differences and the roles of sex steroids in apoptosis of sexually dimorphic nuclei of the preoptic area in postnatal rats. J Neuroendocrinol 21:370–376CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biochemistry and Cellular and Molecular Biology and NeuroNet Research CenterUniversity of TennesseeKnoxvilleUSA
  2. 2.UT-ORNL Graduate School of Genome Science and Technology ProgramUniversity of TennesseeKnoxvilleUSA

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