scholarly journals RNAi screen in Tribolium reveals involvement of F-BAR proteins in myoblast fusion and visceral muscle morphogenesis in insects

2018 ◽  
Author(s):  
Dorothea Schultheis ◽  
Jonas Schwirz ◽  
Manfred Frasch
Keyword(s):  
Large Scale ◽  
Actin Polymerization ◽  
Muscle Development ◽  
Myoblast Fusion ◽  
Rnai Screen ◽  
Gene Encoding ◽  
Genetic Studies ◽  
Knock Down ◽  
Larval Muscle ◽  
New Gene

AbstractIn a large-scale RNAi screen in Tribolium castaneum for genes with knock-down phenotypes in the larval somatic musculature, one recurring phenotype was the appearance of larval muscle fibers that were significantly thinner than those in control animals. Several of the genes producing this knock-down phenotype corresponded to orthologs of Drosophila genes that are known to participate in myoblast fusion, particularly via their effects on actin polymerization. A new gene previously not implicated in myoblast fusion but displaying a similar thin-muscle knock-down phenotype was the Tribolium ortholog of Nostrin, which encodes an F-BAR and SH3 domain protein. Our genetic studies of Nostrin and Cip4, a gene encoding a structurally related protein, in Drosophila show that the encoded F-BAR proteins jointly contribute to efficient myoblast fusion during larval muscle development. Together with the F-Bar protein Syndapin they are also required for normal embryonic midgut morphogenesis. In addition, Cip4 is required together with Nostrin during the profound remodeling of the midgut visceral musculature during metamorphosis. We propose that these F-Bar proteins help govern proper morphogenesis particularly of the longitudinal midgut muscles during metamorphosis.

Identification of the crucial molecular events during the large-scale myoblast fusion in sheep

2014 ◽  
Vol 46 (12) ◽  
pp. 429-440 ◽  
Author(s):  
Caihong Wei ◽  
Li Li ◽  
Hongwei Su ◽  
Lingyang Xu ◽  
Jian Lu ◽  
...  
Keyword(s):  
Large Scale ◽  
Molecular Mechanisms ◽  
Muscle Development ◽  
Target Genes ◽  
Myogenic Differentiation ◽  
Myoblast Fusion ◽  
Myoblast Differentiation ◽  
Pcr Analysis ◽  
Fetal Sheep ◽  
Fetal Muscle

It is well known that in sheep most myofibers are formed before birth; however, the crucial myogenic stage and the cellular and molecular mechanisms underpinning phenotypic variation of fetal muscle development remain to be ascertained. We used histological, microarray, and quantitative real-time PCR (qPCR) methods to examine the developmental characteristics of fetal muscle at 70, 85, 100, 120, and 135 days of gestation in sheep. We show that day 100 is an important checkpoint for change in muscle transcriptome and histomorphology in fetal sheep and that the period of 85–100 days is the vital developmental stage for large-scale myoblast fusion. Furthermore, we identified the cis-regulatory motifs for E2F1 or MEF2A in a list of decreasingly or increasingly expressed genes between 85 and 100 days, respectively. Further analysis demonstrated that the mRNA and phosphorylated protein levels of E2F1 and MEF2A significantly declined with myogenic progression in vivo and in vitro. qRT-PCR analysis indicated that PI3K and FST, as targets of E2F1, may be involved in myoblast differentiation and fusion and that downregulation of MEF2A contributes to transition of myofiber types by differential regulation of the target genes involved at the stage of 85–100 days. We clarify for the first time the timing of myofiber proliferation and development during gestation in sheep, which would be beneficial to meat sheep production. Our findings present a repertoire of gene expression in muscle during large-scale myoblast fusion at transcriptome-wide level, which contributes to elucidate the regulatory network of myogenic differentiation.

scholarly journals A Large Scale Systemic RNAi Screen in the Red Flour Beetle Tribolium castaneum Identifies Novel Genes Involved in Insect Muscle Development

2019 ◽  
pp. g3.200995.2018
Author(s):  
Dorothea Schultheis ◽  
Matthias Weißkopf ◽  
Christoph Schaub ◽  
Salim Ansari ◽  
Van Anh Dao ◽  
...  
Keyword(s):  
Tribolium Castaneum ◽  
Large Scale ◽  
Muscle Development ◽  
Rnai Screen ◽  
Red Flour Beetle ◽  
Novel Genes ◽  
Insect Muscle ◽  
Flour Beetle ◽  
Systemic Rnai

scholarly journals A large scale systemic RNAi screen in the red flour beetle Tribolium castaneum identifies novel genes involved in insect muscle development

2018 ◽  
Author(s):  
Dorothea Schultheis ◽  
Matthias Weißkopf ◽  
Christoph Schaub ◽  
Salim Ansari ◽  
Van Anh Dao ◽  
...  
Keyword(s):  
Tribolium Castaneum ◽  
Large Scale ◽  
Muscle Development ◽  
Rnai Screen ◽  
Red Flour Beetle ◽  
Larval Body ◽  
Insect Muscle ◽  
Flour Beetle ◽  
Systemic Rnai ◽  
Screening Platform

AbstractAlthough muscle development has been widely studied in Drosophila melanogaster there are still many gaps in our knowledge, and it is not known to which extent this knowledge can be transferred to other insects. To help in closing these gaps we participated in a large-scale RNAi screen that used the red flour beetle, Tribolium castaneum, as a screening platform. The effects of systemic RNAi were screened upon double-stranded RNA injections into appropriate muscle-EGFP tester strains. Injections into pupae were followed by the analysis of the late embryonic/early larval muscle patterns, and injections into larvae by the analysis of the adult thoracic muscle patterns. Herein we describe the results of the first-pass screens with pupal and larval injections, which covered ~8,500 and ~5,000 genes, respectively, of a total of ~16,500 genes of the Tribolium genome. Apart from many genes known from Drosophila as regulators of muscle development, a collection of genes previously unconnected to muscle development yielded phenotypes in larval body wall and leg muscles as well as in indirect flight muscles. We then present the main candidates from the pupal injection screen that remained after being processed through a series of verification and selection steps. Further, we discuss why distinct though overlapping sets of genes are revealed by the Drosophila and Tribolium screening approaches.

scholarly journals Large-scale RNAi screen identified Dhpr as a regulator of mitochondrial morphology and tissue homeostasis

2019 ◽  
Vol 5 (9) ◽  
pp. eaax0365 ◽  
Author(s):  
Jia Zhou ◽  
Lingna Xu ◽  
Xiuying Duan ◽  
Wei Liu ◽  
Xiaocui Zhao ◽  
...  
Keyword(s):  
Large Scale ◽  
Complex Analysis ◽  
Rnai Screen ◽  
Tissue Homeostasis ◽  
Mitochondrial Morphology ◽  
Gene Encoding ◽  
Chain Complexes ◽  
Transport Chain ◽  
Gradual Loss

Mitochondria are highly dynamic organelles. Through a large-scale in vivo RNA interference (RNAi) screen that covered around a quarter of the Drosophila melanogaster genes (4000 genes), we identified 578 genes whose knockdown led to aberrant shapes or distributions of mitochondria. The complex analysis revealed that knockdown of the subunits of proteasomes, spliceosomes, and the electron transport chain complexes could severely affect mitochondrial morphology. The loss of Dhpr, a gene encoding an enzyme catalyzing tetrahydrobiopterin regeneration, leads to a reduction in the numbers of tyrosine hydroxylase neurons, shorter lifespan, and gradual loss of muscle integrity and climbing ability. The affected mitochondria in Dhpr mutants are swollen and have fewer cristae, probably due to lower levels of Drp1 S-nitrosylation. Overexpression of Drp1, but not of S-nitrosylation–defective Drp1, rescued Dhpr RNAi-induced mitochondrial defects. We propose that Dhpr regulates mitochondrial morphology and tissue homeostasis by modulating S-nitrosylation of Drp1.

The Use of Medical Record Linkage for Population and Genetic Studies

1969 ◽  
Vol 08 (01) ◽  
pp. 07-11 ◽  
Author(s):  
H. B. Newcombe
Keyword(s):  
Record Linkage ◽  
Large Scale ◽  
Medical Record Linkage ◽  
Canadian Province ◽  
Genetic Studies ◽  
Parental Characteristics ◽  
Family Histories ◽  
The Family ◽  
Large Populations ◽  
Machine Readable

Methods are described for deriving personal and family histories of birth, marriage, procreation, ill health and death, for large populations, from existing civil registrations of vital events and the routine records of ill health. Computers have been used to group together and »link« the separately derived records pertaining to successive events in the lives of the same individuals and families, rapidly and on a large scale. Most of the records employed are already available as machine readable punchcards and magnetic tapes, for statistical and administrative purposes, and only minor modifications have been made to the manner in which these are produced.As applied to the population of the Canadian province of British Columbia (currently about 2 million people) these methods have already yielded substantial information on the risks of disease: a) in the population, b) in relation to various parental characteristics, and c) as correlated with previous occurrences in the family histories.

scholarly journals Reconstitution of contractile actomyosin rings in vesicles

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Thomas Litschel ◽  
Charlotte F. Kelley ◽  
Danielle Holz ◽  
Maral Adeli Koudehi ◽  
Sven K. Vogel ◽  
...  
Keyword(s):  
Large Scale ◽  
Actin Polymerization ◽  
Living Cells ◽  
Actin Ring ◽  
Actin Networks ◽  
Mechanical Transformation ◽  
Myosin Motors ◽  
Spherical Vesicles ◽  
Theoretical Considerations ◽  
Encapsulation Methods

AbstractOne of the grand challenges of bottom-up synthetic biology is the development of minimal machineries for cell division. The mechanical transformation of large-scale compartments, such as Giant Unilamellar Vesicles (GUVs), requires the geometry-specific coordination of active elements, several orders of magnitude larger than the molecular scale. Of all cytoskeletal structures, large-scale actomyosin rings appear to be the most promising cellular elements to accomplish this task. Here, we have adopted advanced encapsulation methods to study bundled actin filaments in GUVs and compare our results with theoretical modeling. By changing few key parameters, actin polymerization can be differentiated to resemble various types of networks in living cells. Importantly, we find membrane binding to be crucial for the robust condensation into a single actin ring in spherical vesicles, as predicted by theoretical considerations. Upon force generation by ATP-driven myosin motors, these ring-like actin structures contract and locally constrict the vesicle, forming furrow-like deformations. On the other hand, cortex-like actin networks are shown to induce and stabilize deformations from spherical shapes.

scholarly journals Mechanisms of myoblast fusion during muscle development

2015 ◽  
Vol 32 ◽  
pp. 162-170 ◽  
Author(s):  
Ji Hoon Kim ◽  
Peng Jin ◽  
Rui Duan ◽  
Elizabeth H Chen
Keyword(s):  
Muscle Development ◽  
Myoblast Fusion
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