A large-scale transgenic RNAi screen identifies transcription factors that modulate myofiber size in Drosophila

Graça, Flávia A.; Sheffield, Natalie; Puppa, Melissa; Finkelstein, David; Hunt, Liam C.; Demontis, Fabio

doi:10.1371/journal.pgen.1009926

Cited by 14 publications

(11 citation statements)

References 88 publications

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“…Overall, we followed an experimental pipeline (Fig. 1a, b ) similar to the one we recently employed to screen for ubiquitin ligases 60 and transcription factors 66 . Specifically, the UAS/Gal4 system 67 , the skeletal muscle-specific Mef2-Gal4 driver 59 , 68 , and 508 UAS-RNAi transgenic fly stocks were used to knock down 111 evolutionary-conserved myokines that have strong muscle expression (RNA-seq FPKM values ≥4), and assess their role in myofiber growth (Fig.…”

Section: Resultsmentioning

confidence: 99%

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The myokine Fibcd1 is an endogenous determinant of myofiber size and mitigates cancer-induced myofiber atrophy

et al. 2022

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Decline in skeletal muscle cell size (myofiber atrophy) is a key feature of cancer-induced wasting (cachexia). In particular, atrophy of the diaphragm, the major muscle responsible for breathing, is an important determinant of cancer-associated mortality. However, therapeutic options are limited. Here, we have used Drosophila transgenic screening to identify muscle-secreted factors (myokines) that act as paracrine regulators of myofiber growth. Subsequent testing in mouse myotubes revealed that mouse Fibcd1 is an evolutionary-conserved myokine that preserves myofiber size via ERK signaling. Local administration of recombinant Fibcd1 (rFibcd1) ameliorates cachexia-induced myofiber atrophy in the diaphragm of mice bearing patient-derived melanoma xenografts and LLC carcinomas. Moreover, rFibcd1 impedes cachexia-associated transcriptional changes in the diaphragm. Fibcd1-induced signaling appears to be muscle selective because rFibcd1 increases ERK activity in myotubes but not in several cancer cell lines tested. We propose that rFibcd1 may help reinstate myofiber size in the diaphragm of patients with cancer cachexia.

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Section: Resultsmentioning

confidence: 99%

“…For the screen, we used similar procedures as we recently used 60 , 66 . Specifically, virgin Mef2-Gal4 females (10 flies for each cross) were mated with males (5 flies for each cross) of each RNAi or overexpression stock and transferred to new food tubes every 3 days to avoid overcrowding.…”

Section: Methodsmentioning

confidence: 99%

The myokine Fibcd1 is an endogenous determinant of myofiber size and mitigates cancer-induced myofiber atrophy

et al. 2022

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“…Other circulating factors that may activate proliferation of SC in non-injury states into myofibers include growth hormone, follistatin (a myostatin antagonist), and IGF-1, among others (Forcina et al, 2019;Murach et al, 2021). Additionally, increases in lactate concentrations may act as a signaling molecule for increased proliferation of SCs (Nalbandian et al, 2020), and others have suggested that glycolytic enzymes are necessary for optimal muscle growth, at least in Drosophila models (Graca et al, 2021). However, it is unclear how regulation to either add new nuclei or upregulate existing myonuclear activity is accomplished within muscle fibers (Cramer et al, 2020).…”

Section: Myonuclear Domain and Its Significance To Muscle Functionmentioning

confidence: 99%

Skeletal muscle and metabolic flexibility in response to changing energy demands in wild birds

2022

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Phenotypically plastic responses of animals to adjust to environmental variation are pervasive. Reversible plasticity (i.e., phenotypic flexibility), where adult phenotypes can be reversibly altered according to prevailing environmental conditions, allow for better matching of phenotypes to the environment and can generate fitness benefits but may also be associated with costs that trade-off with capacity for flexibility. Here, we review the literature on avian metabolic and muscle plasticity in response to season, temperature, migration and experimental manipulation of flight costs, and employ an integrative approach to explore the phenotypic flexibility of metabolic rates and skeletal muscle in wild birds. Basal (minimum maintenance metabolic rate) and summit (maximum cold-induced metabolic rate) metabolic rates are flexible traits in birds, typically increasing with increasing energy demands. Because skeletal muscles are important for energy use at the organismal level, especially to maximum rates of energy use during exercise or shivering thermogenesis, we consider flexibility of skeletal muscle at the tissue and ultrastructural levels in response to variations in the thermal environment and in workloads due to flight exercise. We also examine two major muscle remodeling regulatory pathways: myostatin and insulin-like growth factor -1 (IGF-1). Changes in myostatin and IGF-1 pathways are sometimes, but not always, regulated in a manner consistent with metabolic rate and muscle mass flexibility in response to changing energy demands in wild birds, but few studies have examined such variation so additional study is needed to fully understand roles for these pathways in regulating metabolic flexibility in birds. Muscle ultrastrutural variation in terms of muscle fiber diameter and associated myonuclear domain (MND) in birds is plastic and highly responsive to thermal variation and increases in workload, however, only a few studies have examined ultrastructural flexibility in avian muscle. Additionally, the relationship between myostatin, IGF-1, and satellite cell (SC) proliferation as it relates to avian muscle flexibility has not been addressed in birds and represents a promising avenue for future study.

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“…In vivo RNAi-based genetic screens have proved very successful in identifying novel genes involved in developmental and pathologic processes in a variety of Drosophila somatic tissues when expressed using tissue-specific Gal4 driver lines (Graca et al , 2021; Lesch et al , 2010; Neely et al , 2010; Pletcher et al , 2019; Port et al , 2011; Rotelli et al , 2019; Rylee et al , 2022; Saj et al , 2010; Zeng et al , 2015; Zhou et al , 2019). Hence, we embarked on a candidate genetic screen aimed at identifying factors controlling genome stability.…”

Section: Resultsmentioning

confidence: 99%

DrosophilapVALIUM10 TRiP RNAi lines cause undesired silencing of Gateway-based transgenes

Stanković

Csordás

Uhlířová

2022

Preprint

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SUMMARYPost-transcriptional gene silencing using double-stranded RNA has revolutionized the field of functional genetics, allowing fast and easy disruption of gene function in various organisms. In Drosophila, many transgenic RNAi lines have been generated in large-scale efforts, including the Drosophila Transgenic RNAi Project (TRiP), to facilitate in vivo knockdown of virtually any Drosophila gene with spatial and temporal resolution. The available transgenic RNAi lines represent a fundamental resource for the fly community, providing an unprecedented opportunity to address a vast range of biological questions relevant to basic and biomedical research fields. However, caution should be applied regarding the efficiency and specificity of the RNAi approach. Here, we demonstrate that pVALIUM10-based RNAi lines, representing ∼11% of the total TRiP collection, cause unintended silencing of transgenes expressed from Gateway destination vectors generated via site-specific recombination. The silencing is mediated by targeting attB1 and attB2 sequences generated in the recombination reaction and included in the transcribed mRNA. Deleting these attB sites from the Gateway expression vector prevents silencing and restores expected transgene expression.

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A large-scale transgenic RNAi screen identifies transcription factors that modulate myofiber size in Drosophila

Cited by 14 publications

References 88 publications

The myokine Fibcd1 is an endogenous determinant of myofiber size and mitigates cancer-induced myofiber atrophy

The myokine Fibcd1 is an endogenous determinant of myofiber size and mitigates cancer-induced myofiber atrophy

Skeletal muscle and metabolic flexibility in response to changing energy demands in wild birds

DrosophilapVALIUM10 TRiP RNAi lines cause undesired silencing of Gateway-based transgenes

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