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Two-factor specification of apoptosis: TGF-β signaling acts cooperatively with ecdysone signaling to induce cell- and stage-specific apoptosis of larval neurons during metamorphosis in Drosophila melanogaster

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Abstract

Developmentally regulated programmed cell death (PCD) is one of the key cellular events for precise controlling of neuronal population during postembryonic development of the central nervous system. Previously we have shown that a group of corazonin-producing peptidergic neurons (vCrz) undergo apoptosis in response to ecdysone signaling via ecdysone receptor (EcR)-B isoforms and Ultraspiracle during early phase of metamorphosis. Further utilizing genetic, transgenic, and mosaic analyses, we have found that TGF-β signaling mediated by a glia-produced ligand, Myoglianin, type-I receptor Baboon (particularly Babo-A isoform) and dSmad2, is also required autonomously for PCD of the vCrz neurons. Our studies show that TGF-β signaling is not acting epistatically to EcR or vice versa. We also show that ectopic expression of a constitutively active phosphomimetic form of dSmad2 (dSmad2PM) is capable of inducing premature death of vCrz neurons in larva but not other larval neurons. Intriguingly, the dSmad2PM-mediated killing is completely suppressed by coexpression of a dominant-negative form of EcR (EcRDN), suggesting that EcR function is required for the proapoptotic dSmad2PM function. Based on these data, we suggest that TGF-β and ecdysone signaling pathways act cooperatively to induce vCrz neuronal PCD. We propose that this type of two-factor authentication is a key developmental strategy to ensure the timely PCD of specific larval neurons during metamorphosis.

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Acknowledgements

We are greatly indebted to Michael O’Connor (Univ. of Minnesota) for the provision of numerous fly strains used here. We also thank T. Lee (Janelia Farm), S. Robinow (Univ. of Hawaii), P. Cherbas (Indiana Univ.), and B. Hay (Caltech) for their kind donation of fly lines, and an anonymous reviewer for the helpful suggestions. The work was supported in part by NSF Grant (IOS-0919797), NIH Grant (R15-GM114741) and by Hunsicker research incentive award (Univ. of Tennessee).

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  1. Zixing Wang and Gyunghee Lee have contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Cellular and Molecular Biology and NeuroNet Research Center, University of Tennessee, Knoxville, TN, 37996, USA

    Gyunghee Lee & Jae H. Park

  2. UT-ORNL Graduate School of Genome Science and Technology Program, University of Tennessee, Knoxville, TN, 37996, USA

    Zixing Wang & Jae H. Park

  3. Undergraduate program in Neuroscience, University of Tennessee, Knoxville, TN, 37996, USA

    Richard Vuong

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  1. Zixing Wang
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Correspondence to Jae H. Park.

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Supplemental Fig. 1 Ectopic expression of dSmad2PM did not kill larval neurons expressing other peptidergic neurons. a Control CCAP neuropeptide-expressing larval neurons in WL3 (n = 14, genotype: UAS-mCD8GFP/+; ccap-gal4/+). b Ectopic expression of dSmad2PM in CCAP neurons did not kill them (n = 7, genotypes: UAS-mCD8GFP/+; UAS-dSmad2PM/+; ccap-gal4/+). c DILP2-expressing peptidergic neurons in the larval brain at WL3 (n = 5, genotypes: UAS-mCD8GFP/+; dilp2-gal4/+). d Ectopic expression of dSmad2PM did not kill DILP2 neurons (n = 3, genotype: UAS-mCD8GFP/+; dilp2-Gal4/UAS-dSmad2PM). e MIP-expressing peptidergic larval neurons in the CNS of WL3 (n = 3, UAS-mCD8GFP/+; mip-gal4/+). f Ectopic expression of dSmad2PM did not kill MIP-neurons (n = 3, genotype: UAS-mCD8GFP/+; UAS-dSmad2PM/+; mip-gal4). Scale bars, 100 µm (A), 50 µm (C).

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Wang, Z., Lee, G., Vuong, R. et al. Two-factor specification of apoptosis: TGF-β signaling acts cooperatively with ecdysone signaling to induce cell- and stage-specific apoptosis of larval neurons during metamorphosis in Drosophila melanogaster. Apoptosis 24, 972–989 (2019). https://doi.org/10.1007/s10495-019-01574-4

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