An RNAi based screen in Drosophila larvae identifies fascin as a regulator of myoblast fusion and myotendinous junction structure
Abstract Background A strength of Drosophila as a model system is its utility as a tool to screen for novel regulators of various functional and developmental processes. However, the utility of Drosophila as a screening tool is dependent on the speed and simplicity of the assay used. Methods Here, w...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-04-01
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| Series: | Skeletal Muscle |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13395-018-0159-9 |
| _version_ | 1818137376619233280 |
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| author | Jaclyn M. Camuglia Torrey R. Mandigo Richard Moschella Jenna Mark Christine H. Hudson Derek Sheen Eric S. Folker |
| author_facet | Jaclyn M. Camuglia Torrey R. Mandigo Richard Moschella Jenna Mark Christine H. Hudson Derek Sheen Eric S. Folker |
| author_sort | Jaclyn M. Camuglia |
| collection | DOAJ |
| description | Abstract Background A strength of Drosophila as a model system is its utility as a tool to screen for novel regulators of various functional and developmental processes. However, the utility of Drosophila as a screening tool is dependent on the speed and simplicity of the assay used. Methods Here, we use larval locomotion as an assay to identify novel regulators of skeletal muscle function. We combined this assay with muscle-specific depletion of 82 genes to identify genes that impact muscle function by their expression in muscle cells. The data from the screen were supported with characterization of the muscle pattern in embryos and larvae that had disrupted expression of the strongest hit from the screen. Results With this assay, we showed that 12/82 tested genes regulate muscle function. Intriguingly, the disruption of five genes caused an increase in muscle function, illustrating that mechanisms that reduce muscle function exist and that the larval locomotion assay is sufficiently quantitative to identify conditions that both increase and decrease muscle function. We extended the data from this screen and tested the mechanism by which the strongest hit, fascin, impacted muscle function. Compared to controls, animals in which fascin expression was disrupted with either a mutant allele or muscle-specific expression of RNAi had fewer muscles, smaller muscles, muscles with fewer nuclei, and muscles with disrupted myotendinous junctions. However, expression of RNAi against fascin only after the muscle had finished embryonic development did not recapitulate any of these phenotypes. Conclusions These data suggest that muscle function is reduced due to impaired myoblast fusion, muscle growth, and muscle attachment. Together, these data demonstrate the utility of Drosophila larval locomotion as an assay for the identification of novel regulators of muscle development and implicate fascin as necessary for embryonic muscle development. |
| first_indexed | 2024-12-11T09:55:19Z |
| format | Article |
| id | doaj.art-ea00455abbad4f00b3885d54b237498d |
| institution | Directory Open Access Journal |
| issn | 2044-5040 |
| language | English |
| last_indexed | 2024-12-11T09:55:19Z |
| publishDate | 2018-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Skeletal Muscle |
| spelling | doaj.art-ea00455abbad4f00b3885d54b237498d2022-12-22T01:12:17ZengBMCSkeletal Muscle2044-50402018-04-018111310.1186/s13395-018-0159-9An RNAi based screen in Drosophila larvae identifies fascin as a regulator of myoblast fusion and myotendinous junction structureJaclyn M. Camuglia0Torrey R. Mandigo1Richard Moschella2Jenna Mark3Christine H. Hudson4Derek Sheen5Eric S. Folker6Biology Department, Boston CollegeBiology Department, Boston CollegeBiology Department, Boston CollegeBiology Department, Boston CollegeBiology Department, Boston CollegeBiology Department, Boston CollegeBiology Department, Boston CollegeAbstract Background A strength of Drosophila as a model system is its utility as a tool to screen for novel regulators of various functional and developmental processes. However, the utility of Drosophila as a screening tool is dependent on the speed and simplicity of the assay used. Methods Here, we use larval locomotion as an assay to identify novel regulators of skeletal muscle function. We combined this assay with muscle-specific depletion of 82 genes to identify genes that impact muscle function by their expression in muscle cells. The data from the screen were supported with characterization of the muscle pattern in embryos and larvae that had disrupted expression of the strongest hit from the screen. Results With this assay, we showed that 12/82 tested genes regulate muscle function. Intriguingly, the disruption of five genes caused an increase in muscle function, illustrating that mechanisms that reduce muscle function exist and that the larval locomotion assay is sufficiently quantitative to identify conditions that both increase and decrease muscle function. We extended the data from this screen and tested the mechanism by which the strongest hit, fascin, impacted muscle function. Compared to controls, animals in which fascin expression was disrupted with either a mutant allele or muscle-specific expression of RNAi had fewer muscles, smaller muscles, muscles with fewer nuclei, and muscles with disrupted myotendinous junctions. However, expression of RNAi against fascin only after the muscle had finished embryonic development did not recapitulate any of these phenotypes. Conclusions These data suggest that muscle function is reduced due to impaired myoblast fusion, muscle growth, and muscle attachment. Together, these data demonstrate the utility of Drosophila larval locomotion as an assay for the identification of novel regulators of muscle development and implicate fascin as necessary for embryonic muscle development.http://link.springer.com/article/10.1186/s13395-018-0159-9Nuclear movementDrosophilaMyoblast fusionMyotendinous junctionFascinMyogenesis |
| spellingShingle | Jaclyn M. Camuglia Torrey R. Mandigo Richard Moschella Jenna Mark Christine H. Hudson Derek Sheen Eric S. Folker An RNAi based screen in Drosophila larvae identifies fascin as a regulator of myoblast fusion and myotendinous junction structure Skeletal Muscle Nuclear movement Drosophila Myoblast fusion Myotendinous junction Fascin Myogenesis |
| title | An RNAi based screen in Drosophila larvae identifies fascin as a regulator of myoblast fusion and myotendinous junction structure |
| title_full | An RNAi based screen in Drosophila larvae identifies fascin as a regulator of myoblast fusion and myotendinous junction structure |
| title_fullStr | An RNAi based screen in Drosophila larvae identifies fascin as a regulator of myoblast fusion and myotendinous junction structure |
| title_full_unstemmed | An RNAi based screen in Drosophila larvae identifies fascin as a regulator of myoblast fusion and myotendinous junction structure |
| title_short | An RNAi based screen in Drosophila larvae identifies fascin as a regulator of myoblast fusion and myotendinous junction structure |
| title_sort | rnai based screen in drosophila larvae identifies fascin as a regulator of myoblast fusion and myotendinous junction structure |
| topic | Nuclear movement Drosophila Myoblast fusion Myotendinous junction Fascin Myogenesis |
| url | http://link.springer.com/article/10.1186/s13395-018-0159-9 |
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