The miR-30 MicroRNA Family Targets smoothened to Regulate Hedgehog Signalling in Zebrafish Early Muscle Development
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{"title"=>"The miR-30 MicroRNA Family Targets smoothened to Regulate Hedgehog Signalling in Zebrafish Early Muscle Development", "type"=>"journal", "authors"=>[{"first_name"=>"Ami", "last_name"=>"Ketley", "scopus_author_id"=>"55131440100"}, {"first_name"=>"Anne", "last_name"=>"Warren", "scopus_author_id"=>"23101614100"}, {"first_name"=>"Emily", "last_name"=>"Holmes", "scopus_author_id"=>"54942656900"}, {"first_name"=>"Martin", "last_name"=>"Gering", "scopus_author_id"=>"6602377267"}, {"first_name"=>"A. Aziz", "last_name"=>"Aboobaker", "scopus_author_id"=>"6507411601"}, {"first_name"=>"J. David", "last_name"=>"Brook", "scopus_author_id"=>"55663196500"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84878754811", "doi"=>"10.1371/journal.pone.0065170", "pui"=>"369075518", "issn"=>"19326203", "pmid"=>"23755189", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "scopus"=>"2-s2.0-84878754811"}, "id"=>"f37816ef-c6c6-34b5-b498-6ef10507d06a", "abstract"=>"The importance of microRNAs in development is now widely accepted. However, identifying the specific targets of individual microRNAs and understanding their biological significance remains a major challenge. We have used the zebrafish model system to evaluate the expression and function of microRNAs potentially involved in muscle development and study their interaction with predicted target genes. We altered expression of the miR-30 microRNA family and generated phenotypes that mimicked misregulation of the Hedgehog pathway. Inhibition of the miR-30 family increases activity of the pathway, resulting in elevated ptc1 expression and increased numbers of superficial slow-muscle fibres. We show that the transmembrane receptor smoothened is a target of this microRNA family. Our results indicate that fine coordination of smoothened activity by the miR-30 family allows the correct specification and differentiation of distinct muscle cell types during zebrafish embryonic development.", "link"=>"http://www.mendeley.com/research/mir30-microrna-family-targets-smoothened-regulate-hedgehog-signalling-zebrafish-early-muscle-develop", "reader_count"=>33, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>4, "Researcher"=>7, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>1, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>1, "Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>4, "Researcher"=>7, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>1, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>1, "Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Biochemistry, Genetics and Molecular Biology"=>7, "Medicine and Dentistry"=>3, "Agricultural and Biological Sciences"=>18, "Psychology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Psychology"=>{"Psychology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>18}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>7}, "Unspecified"=>{"Unspecified"=>4}}, "reader_count_by_country"=>{"Argentina"=>1, "Spain"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1074804"], "description"=>"<p>The miR-30 family shows extremely high sequence similarity and an identical seed sequence, as highlighted by the red box.</p>", "links"=>[], "tags"=>["developmental biology", "Cell differentiation", "Cell fate determination", "genetics", "Molecular genetics", "Gene regulation", "Gene function", "Model organisms", "Animal models", "zebrafish", "Molecular cell biology", "gene expression", "Signal transduction", "mir-30", "microrna", "shows", "overlapping", "patterns", "embryonic"], "article_id"=>711985, "categories"=>["Biological Sciences"], "users"=>["Ami Ketley", "Anne Warren", "Emily Holmes", "Martin Gering", "A. Aziz Aboobaker", "J. David Brook"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0065170.g001", "stats"=>{"downloads"=>6, "page_views"=>299, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_miR_30_microRNA_family_shows_high_sequence_similarity_and_overlapping_expression_patterns_throughout_embryonic_development_/711985", "title"=>"The miR-30 microRNA family shows high sequence similarity and overlapping expression patterns throughout embryonic development.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-05 00:33:05"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074805"], "description"=>"<p>Embryo somite structure at 24 hpf is shown (<b>A–D</b>). <i>Ptc1</i> expression analysis was used as a read out of Hh pathway activity, showing elevated levels in miR-30 morpholino and dnPKA treated embryos when compared to wild type embryos (<b>E–H</b>). Slow muscle fibre number was quantified by immunohistochemistry using the S58 antibody (yellow/green) and MF20 staining (red). Embryo sections are orientated dorsal side upwards (<b>I–L</b>). Images are shown of wild type embryos (<b>A, E, I</b>), miR-30 morpholino treated embryos (<b>B, F, J</b>), dnPKA treated embryos (<b>C, G, K</b>) and miR-30 overexpression embryos (<b>D, H, L</b>).</p>", "links"=>[], "tags"=>["developmental biology", "Cell differentiation", "Cell fate determination", "genetics", "Molecular genetics", "Gene regulation", "Gene function", "Model organisms", "Animal models", "zebrafish", "Molecular cell biology", "gene expression", "Signal transduction", "mir-30", "embryonic", "hh", "pathway"], "article_id"=>711986, "categories"=>["Biological Sciences"], "users"=>["Ami Ketley", "Anne Warren", "Emily Holmes", "Martin Gering", "A. Aziz Aboobaker", "J. David Brook"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0065170.g002", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_miR_30_family_is_required_during_early_embryonic_development_to_regulate_Hh_pathway_activity_/711986", "title"=>"The miR-30 family is required during early embryonic development to regulate Hh pathway activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-05 00:33:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074806"], "description"=>"<p>(<b>A–F</b>) Embryos injected with 3 different GFP reporter mRNAs; (<b>A,B</b>) the GFP ORF plus tandem perfect target sites (GFP-PTS), (<b>C,D</b>) GFP ORF plus the smo 3′UTR sequence (GFP-SMO) and (<b>E,F</b>) the GFP ORF without UTR sequence (GFP- no UTR). Constructs were injected either alone (<b>A,C,E</b>) or with the miR-30 duplex RNA (<b>B,D,F</b>). (<b>G</b>) Western blot validation on lysates of GFP injected embryos with and without the miR-30 duplex (<b>H</b>) Densitometric analysis of GFP protein levels normalised against α-tubulin loading control shows a 54% reduction in GFP-SMO+miR-30 compared to GFP-SMO only embryos. (<b>I</b>) Protein blot analysis of smoothened levels in wild type and miR-30 morpholino knockdown embryos shows an increased level of Smoothened protein. (<b>J</b>) Densitometric analysis of the average change in smoothened protein level in 3 samples of wild type versus miR-30 morpholino treated embryos.</p>", "links"=>[], "tags"=>["developmental biology", "Cell differentiation", "Cell fate determination", "genetics", "Molecular genetics", "Gene regulation", "Gene function", "Model organisms", "Animal models", "zebrafish", "Molecular cell biology", "gene expression", "Signal transduction", "targets", "hedgehog", "transmembrane", "receptor"], "article_id"=>711987, "categories"=>["Biological Sciences"], "users"=>["Ami Ketley", "Anne Warren", "Emily Holmes", "Martin Gering", "A. Aziz Aboobaker", "J. David Brook"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0065170.g003", "stats"=>{"downloads"=>2, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_30_directly_targets_the_3_8242_UTR_of_the_Hedgehog_transmembrane_receptor_smoothened_/711987", "title"=>"miR-30 directly targets the 3′UTR of the Hedgehog transmembrane receptor smoothened.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-05 00:33:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074807"], "description"=>"<p>Ptc1 in situ hybridization shows the level of Hh pathway activity in different embryo treatment types. (<b>A</b>) Wild type embryo, (<b>B</b>) miR-30 overexpression embryo, (<b>C</b>) miR-30 morpholino injected embryo, (<b>D</b>) dnPKA overexpression embryo (<b>E</b>) dnPKA RNA injected with miR-30 duplex (<b>F</b>) <i>Shh</i> overexpression embryo (<b>G</b>) Reduced <i>ptc1</i> expression is seen in <i>Shh</i> overexpression embryos coinjected with miR-30 RNA as miR-30 is able to suppress pathway activity.</p>", "links"=>[], "tags"=>["developmental biology", "Cell differentiation", "Cell fate determination", "genetics", "Molecular genetics", "Gene regulation", "Gene function", "Model organisms", "Animal models", "zebrafish", "Molecular cell biology", "gene expression", "Signal transduction", "ptc1", "reveals", "mir-30", "hh"], "article_id"=>711988, "categories"=>["Biological Sciences"], "users"=>["Ami Ketley", "Anne Warren", "Emily Holmes", "Martin Gering", "A. Aziz Aboobaker", "J. David Brook"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0065170.g004", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_Ptc1_reveals_the_position_of_miR_30_regulation_in_the_Hh_pathway_/711988", "title"=>"Analysis of Ptc1 reveals the position of miR-30 regulation in the Hh pathway.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-05 00:33:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074808"], "description"=>"<p>(<b>A–D</b>) Somite angle analysis in wild type, miR-30 morpholino and smoothened protector morpholino injected embryos. Somite structure of (<b>A</b>) wild type embryos (<b>B</b>) miR-30 morpholino injected embryos (<b>C</b>) smoothened protector morpholino injected embryos. (<b>D</b>) Histogram to show the average somite angle in wild type and treated embryos. (<b>E–M</b>) Cyclopamine treatment causes reversion in somite structure to a more wild type phenotype. Images as shown of wild type embryos (<b>E,F,G</b>), miR-30 morpholino embryos (<b>H,I,J</b>) and miR-30 morpholino embryos treated with cyclopamine (<b>K,L,M</b>). The somite structure of (<b>E</b>) wild type (<b>H</b>) miR-30 morpholino and (<b>K</b>) miR-30 morpholino and cyclopamine treated embryos. (<b>F, I, L</b>) Expression of <i>ptc1</i> is substantially reduced following cyclopamine treatment (<b>L</b>). Embryos are shown with anterior end to the left and dorsal side up. (<b>G,J,M</b>) The expanded band of slow fibres, as stained by S58 antibody, is restored to the wild type distribution following cyclopamine treatment (<b>M</b>). Embryo sections are orientated dorsal side to the left.</p>", "links"=>[], "tags"=>["developmental biology", "Cell differentiation", "Cell fate determination", "genetics", "Molecular genetics", "Gene regulation", "Gene function", "Model organisms", "Animal models", "zebrafish", "Molecular cell biology", "gene expression", "Signal transduction", "acts", "negatively", "smoothened"], "article_id"=>711989, "categories"=>["Biological Sciences"], "users"=>["Ami Ketley", "Anne Warren", "Emily Holmes", "Martin Gering", "A. Aziz Aboobaker", "J. David Brook"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0065170.g005", "stats"=>{"downloads"=>5, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_30_acts_to_negatively_regulate_smoothened_in_developing_embryos_/711989", "title"=>"miR-30 acts to negatively regulate smoothened in developing embryos.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-06-05 00:33:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/1074809", "https://ndownloader.figshare.com/files/1074810", "https://ndownloader.figshare.com/files/1074811", "https://ndownloader.figshare.com/files/1074812", "https://ndownloader.figshare.com/files/1074813"], "description"=>"<div><p>The importance of microRNAs in development is now widely accepted. However, identifying the specific targets of individual microRNAs and understanding their biological significance remains a major challenge. We have used the zebrafish model system to evaluate the expression and function of microRNAs potentially involved in muscle development and study their interaction with predicted target genes. We altered expression of the miR-30 microRNA family and generated phenotypes that mimicked misregulation of the Hedgehog pathway. Inhibition of the miR-30 family increases activity of the pathway, resulting in elevated <i>ptc1</i> expression and increased numbers of superficial slow-muscle fibres. We show that the transmembrane receptor <i>smoothened</i> is a target of this microRNA family. Our results indicate that fine coordination of smoothened activity by the miR-30 family allows the correct specification and differentiation of distinct muscle cell types during zebrafish embryonic development.</p></div>", "links"=>[], "tags"=>["developmental biology", "Cell differentiation", "Cell fate determination", "genetics", "Molecular genetics", "Gene regulation", "Gene function", "Model organisms", "Animal models", "zebrafish", "Molecular cell biology", "gene expression", "Signal transduction", "mir-30", "microrna", "targets", "hedgehog", "signalling"], "article_id"=>711990, "categories"=>["Biological Sciences"], "users"=>["Ami Ketley", "Anne Warren", "Emily Holmes", "Martin Gering", "A. Aziz Aboobaker", "J. David Brook"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0065170.s001", "https://dx.doi.org/10.1371/journal.pone.0065170.s002", "https://dx.doi.org/10.1371/journal.pone.0065170.s003", "https://dx.doi.org/10.1371/journal.pone.0065170.s004", "https://dx.doi.org/10.1371/journal.pone.0065170.s005"], "stats"=>{"downloads"=>5, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/The_miR_30_MicroRNA_Family_Targets_smoothened_to_Regulate_Hedgehog_Signalling_in_Zebrafish_Early_Muscle_Development/711990", "title"=>"The miR-30 MicroRNA Family Targets <i>smoothened</i> to Regulate Hedgehog Signalling in Zebrafish Early Muscle Development", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-06-05 00:33:10"}

PMC Usage Stats | Further Information

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Relative Metric

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