Guidelines for morpholino use in zebrafish

Stainier, Didier Y. R.; Raz, Erez; Lawson, Nathan D.; Ekker, Stephen C.; Burdine, Rebecca D.; Eisen, Judith S; Ingham, Philip W.; Schulte‐Merker, Stefan; Yelon, Deborah; Weinstein, Brant M.; Mullins, Mary C.; Wilson, Stephen W.; Ramakrishnan, Lalita; Amacher, Sharon L.; Neuhauss, Stephan C.F.; Meng, Anming; Mochizuki, Naoki; Panula, Pertti; Moens, Cecilia B.

doi:10.1371/journal.pgen.1007000

Cited by 284 publications

(266 citation statements)

References 10 publications

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“…This study allowed us to define the loss of function properties of one DEPDC5 missense mutation alongside one nonsense mutation found in epileptic patients in vivo. Furthermore, in this report, we have performed all the necessary controls to validate the phenotype derived by knockdown through antisense morpholino oligonucleotides, including the use of both splice and ATG‐blocking oligonucleotides, phenotypic rescue by the WT human cDNA, and injection of mismatch‐oligo controls 32, 33…”

Section: Discussionmentioning

confidence: 99%

Depdc5 knockdown causes mTOR‐dependent motor hyperactivity in zebrafish

Calbiac

Dabacan²,

Marsan

et al. 2018

Ann Clin Transl Neurol

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Objective DEPDC5 was identified as a major genetic cause of focal epilepsy with deleterious mutations found in a wide range of inherited forms of focal epilepsy, associated with malformation of cortical development in certain cases. Identification of frameshift, truncation, and deletion mutations implicates haploinsufficiency of DEPDC5 in the etiology of focal epilepsy. DEPDC5 is a component of the GATOR1 complex, acting as a negative regulator of mTOR signaling.MethodsZebrafish represents a vertebrate model suitable for genetic analysis and drug screening in epilepsy‐related disorders. In this study, we defined the expression of depdc5 during development and established an epilepsy model with reduced Depdc5 expression.ResultsHere we report a zebrafish model of Depdc5 loss‐of‐function that displays a measurable behavioral phenotype, including hyperkinesia, circular swimming, and increased neuronal activity. These phenotypic features persisted throughout embryonic development and were significantly reduced upon treatment with the mTORC1 inhibitor, rapamycin, as well as overexpression of human WT DEPDC5 transcript. No phenotypic rescue was obtained upon expression of epilepsy‐associated DEPDC5 mutations (p.Arg487* and p.Arg485Gln), indicating that these mutations cause a loss of function of the protein.InterpretationThis study demonstrates that Depdc5 knockdown leads to early‐onset phenotypic features related to motor and neuronal hyperactivity. Restoration of phenotypic features by WT but not epilepsy‐associated Depdc5 mutants, as well as by mTORC1 inhibition confirm the role of Depdc5 in the mTORC1‐dependent molecular cascades, defining this pathway as a potential therapeutic target for DEPDC5‐inherited forms of focal epilepsy.

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Section: Discussionmentioning

confidence: 99%

Depdc5 knockdown causes mTOR‐dependent motor hyperactivity in zebrafish

Calbiac

Dabacan²,

Marsan

et al. 2018

Ann Clin Transl Neurol

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“…10, 2018; combinations when rhombomeric regions were analyzed ( Figure 5H; Table 3). The penetrance of the phenotype is less profound in compound mutants, most probably due to genetic compensation as previously described (Rossi et al, 2015;Stainier et al, 2017). To study whether this is a cell-autonomous effect, we specifically decreased All rights reserved.…”

Section: Yapmentioning

confidence: 94%

Yap/Taz-TEAD ACTIVITY LINKS MECHANICAL CUES TO CELL PROGENITOR BEHAVIOR DURING HINDBRAIN SEGMENTATION

Voltes

Dingare

et al. 2018

Preprint

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“…MOs are specially modified stable antisense oligomers designed to block either gene translation or gene splicing when injected into embryos [275]. In the absence of reverse genetic tools, MO-based experiments rapidly gained popularity in the fish community.…”

Section: Loss-of-function Approaches In the Developing Zebrafishmentioning

confidence: 99%

Consensus guidelines for the use and interpretation of angiogenesis assays

et al. 2018

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The formation of new blood vessels, or angiogenesis, is a complex process that plays important roles in growth and development, tissue and organ regeneration, as well as numerous pathological conditions. Angiogenesis undergoes multiple discrete steps that can be individually evaluated and quantified by a large number of bioassays. These independent assessments hold advantages but also have limitations. This article describes in vivo, ex vivo, and in vitro bioassays that are available for the evaluation of angiogenesis and highlights critical aspects that are relevant for their execution and proper interpretation. As such, this collaborative work is the first edition of consensus guidelines on angiogenesis bioassays to serve for current and future reference.

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Guidelines for morpholino use in zebrafish

Cited by 284 publications

References 10 publications

Depdc5 knockdown causes mTOR‐dependent motor hyperactivity in zebrafish

Depdc5 knockdown causes mTOR‐dependent motor hyperactivity in zebrafish

Yap/Taz-TEAD ACTIVITY LINKS MECHANICAL CUES TO CELL PROGENITOR BEHAVIOR DURING HINDBRAIN SEGMENTATION

Consensus guidelines for the use and interpretation of angiogenesis assays

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