Live imaging of muscle histolysis in Drosophila metamorphosis

Background: The contribution of programmed cell death (PCD) to muscle wasting disorders remains a matter of debate. Drosophila melanogaster metamorphosis offers the opportunity to study muscle cell death in the context of development. Using live cell imaging of the abdomen, two groups of larval mus...

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Main Authors: Kuleesha, Yadav, Puah, Wee Choo, Wasser, Martin
Other Authors: School of Computer Engineering
Format: Article
Language:English
Published: 2016
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Online Access:https://hdl.handle.net/10356/81996
http://hdl.handle.net/10220/41061
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-819962022-02-16T16:26:53Z Live imaging of muscle histolysis in Drosophila metamorphosis Kuleesha, Yadav Puah, Wee Choo Wasser, Martin School of Computer Engineering Live imaging Drosophila Background: The contribution of programmed cell death (PCD) to muscle wasting disorders remains a matter of debate. Drosophila melanogaster metamorphosis offers the opportunity to study muscle cell death in the context of development. Using live cell imaging of the abdomen, two groups of larval muscles can be observed, doomed muscles that undergo histolysis and persistent muscles that are remodelled and survive into adulthood. Method: To identify and characterize genes that control the decision between survival and cell death of muscles, we developed a method comprising in vivo imaging, targeted gene perturbation and time-lapse image analysis. Our approach enabled us to study the cytological and temporal aspects of abnormal cell death phenotypes. Results: In a previous genetic screen for genes controlling muscle size and cell death in metamorphosis, we identified gene perturbations that induced cell death of persistent or inhibit histolysis of doomed larval muscles. RNA interference (RNAi) of the genes encoding the helicase Rm62 and the lysosomal Cathepsin-L homolog Cysteine proteinase 1 (Cp1) caused premature cell death of persistent muscle in early and mid-pupation, respectively. Silencing of the transcriptional co-repressor Atrophin inhibited histolysis of doomed muscles. Overexpression of dominant-negative Target of Rapamycin (TOR) delayed the histolysis of a subset of doomed and induced ablation of all persistent muscles. RNAi of AMPKα, which encodes a subunit of the AMPK protein complex that senses AMP and promotes ATP formation, led to loss of attachment and a spherical morphology. None of the perturbations affected the survival of newly formed adult muscles, suggesting that the method is useful to find genes that are crucial for the survival of metabolically challenged muscles, like those undergoing atrophy. The ablation of persistent muscles did not affect eclosion of adult flies. Conclusions: Live imaging is a versatile approach to uncover gene functions that are required for the survival of muscle undergoing remodelling, yet are dispensable for other adult muscles. Our approach promises to identify molecular mechanisms that can explain the resilience of muscles to PCD. ASTAR (Agency for Sci., Tech. and Research, S’pore) Published version 2016-08-04T07:07:13Z 2019-12-06T14:44:26Z 2016-08-04T07:07:13Z 2019-12-06T14:44:26Z 2016 Journal Article Kuleesha, Y., Puah, W. C., & Wasser, M. (2016). Live imaging of muscle histolysis in Drosophila metamorphosis. BMC Developmental Biology, 16, 12-. 1471-213X https://hdl.handle.net/10356/81996 http://hdl.handle.net/10220/41061 10.1186/s12861-016-0113-1 27141974 en BMC Developmental Biology © 2016 Kuleesha et al. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated 18 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Live imaging
Drosophila
spellingShingle Live imaging
Drosophila
Kuleesha, Yadav
Puah, Wee Choo
Wasser, Martin
Live imaging of muscle histolysis in Drosophila metamorphosis
description Background: The contribution of programmed cell death (PCD) to muscle wasting disorders remains a matter of debate. Drosophila melanogaster metamorphosis offers the opportunity to study muscle cell death in the context of development. Using live cell imaging of the abdomen, two groups of larval muscles can be observed, doomed muscles that undergo histolysis and persistent muscles that are remodelled and survive into adulthood. Method: To identify and characterize genes that control the decision between survival and cell death of muscles, we developed a method comprising in vivo imaging, targeted gene perturbation and time-lapse image analysis. Our approach enabled us to study the cytological and temporal aspects of abnormal cell death phenotypes. Results: In a previous genetic screen for genes controlling muscle size and cell death in metamorphosis, we identified gene perturbations that induced cell death of persistent or inhibit histolysis of doomed larval muscles. RNA interference (RNAi) of the genes encoding the helicase Rm62 and the lysosomal Cathepsin-L homolog Cysteine proteinase 1 (Cp1) caused premature cell death of persistent muscle in early and mid-pupation, respectively. Silencing of the transcriptional co-repressor Atrophin inhibited histolysis of doomed muscles. Overexpression of dominant-negative Target of Rapamycin (TOR) delayed the histolysis of a subset of doomed and induced ablation of all persistent muscles. RNAi of AMPKα, which encodes a subunit of the AMPK protein complex that senses AMP and promotes ATP formation, led to loss of attachment and a spherical morphology. None of the perturbations affected the survival of newly formed adult muscles, suggesting that the method is useful to find genes that are crucial for the survival of metabolically challenged muscles, like those undergoing atrophy. The ablation of persistent muscles did not affect eclosion of adult flies. Conclusions: Live imaging is a versatile approach to uncover gene functions that are required for the survival of muscle undergoing remodelling, yet are dispensable for other adult muscles. Our approach promises to identify molecular mechanisms that can explain the resilience of muscles to PCD.
author2 School of Computer Engineering
author_facet School of Computer Engineering
Kuleesha, Yadav
Puah, Wee Choo
Wasser, Martin
format Article
author Kuleesha, Yadav
Puah, Wee Choo
Wasser, Martin
author_sort Kuleesha, Yadav
title Live imaging of muscle histolysis in Drosophila metamorphosis
title_short Live imaging of muscle histolysis in Drosophila metamorphosis
title_full Live imaging of muscle histolysis in Drosophila metamorphosis
title_fullStr Live imaging of muscle histolysis in Drosophila metamorphosis
title_full_unstemmed Live imaging of muscle histolysis in Drosophila metamorphosis
title_sort live imaging of muscle histolysis in drosophila metamorphosis
publishDate 2016
url https://hdl.handle.net/10356/81996
http://hdl.handle.net/10220/41061
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