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{"title"=>"Changing hydrozoan bauplans by silencing Hox-like genes", "type"=>"journal", "authors"=>[{"first_name"=>"Wolfgang", "last_name"=>"Jakob", "scopus_author_id"=>"35738790400"}, {"first_name"=>"Bernd", "last_name"=>"Schierwater", "scopus_author_id"=>"7003327151"}], "year"=>2007, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-40349091870", "pmid"=>"17668071", "sgr"=>"40349091870", "doi"=>"10.1371/journal.pone.0000694", "isbn"=>"1932-6203", "issn"=>"19326203", "pui"=>"351336891"}, "id"=>"09c9e587-c7fe-30ea-821c-3afac63df3bb", "abstract"=>"Regulatory genes of the Antp class have been a major factor for the invention and radiation of animal bauplans. One of the most diverse animal phyla are the Cnidaria, which are close to the root of metazoan life and which often appear in two distinct generations and a remarkable variety of body forms. Hox-like genes have been known to be involved in axial patterning in the Cnidaria and have been suspected to play roles in the genetic control of many of the observed bauplan changes. Unfortunately RNAi mediated gene silencing studies have not been satisfactory for marine invertebrate organisms thus far. No direct evidence supporting Hox-like gene induced bauplan changes in cnidarians have been documented as of yet. Herein, we report a protocol for RNAi transfection of marine invertebrates and demonstrate that knock downs of Hox-like genes in Cnidaria create substantial bauplan alterations, including the formation of multiple oral poles (\"heads\") by Cnox-2 and Cnox-3 inhibition, deformation of the main body axis by Cnox-5 inhibition and duplication of tentacles by Cnox-1 inhibition. All phenotypes observed in the course of the RNAi studies were identical to those obtained by morpholino antisense oligo experiments and are reminiscent of macroevolutionary bauplan changes. The reported protocol will allow routine RNAi studies in marine invertebrates to be established.", "link"=>"http://www.mendeley.com/research/changing-hydrozoan-bauplans-silencing-hoxlike-genes", "reader_count"=>62, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>5, "Researcher"=>24, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>4}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>5, "Researcher"=>24, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>4}, "reader_count_by_subject_area"=>{"Environmental Science"=>3, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>51, "Medicine and Dentistry"=>1, "Computer Science"=>1, "Earth and Planetary Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Earth and Planetary Sciences"=>{"Earth and Planetary Sciences"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>51}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Environmental Science"=>{"Environmental Science"=>3}}, "reader_count_by_country"=>{"United States"=>3, "Norway"=>1, "Egypt"=>1, "Brazil"=>1, "United Kingdom"=>1, "Germany"=>1, "Russia"=>1}, "group_count"=>1}

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• http://doi.org/10.7554/eLife.01939
• http://doi.org/10.1093/icb/ics098
• http://doi.org/10.4061/2011/782187
• http://doi.org/10.1016/j.ydbio.2009.05.563
• http://doi.org/10.1371/journal.pone.0112326
• http://doi.org/10.1186/1472-6750-11-67
• http://doi.org/10.1371/journal.pbio.1000020
• http://doi.org/10.1371/journal.pone.0034682
• http://doi.org/10.1016/j.gde.2016.07.006
• http://doi.org/10.1093/gbe/evv153
• http://doi.org/10.1007/s10126-010-9295-x
• http://doi.org/10.1038/s41467-018-04184-x
• http://doi.org/10.1371/journal.pone.0004231
• http://doi.org/10.1016/S1672-0229(11)60010-7

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• {"url"=>"http%3A%2F%2Fjournals.plos.org%2Fplosone%2Farticle%3Fid%3D10.1371%252Fjournal.pone.0000694", "share_count"=>0, "like_count"=>0, "comment_count"=>0, "click_count"=>0, "total_count"=>0}

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• {"type"=>"COMMENT", "annotationUri"=>"info:doi/10.1371/annotation/a775f1fd-a31c-49f8-bb73-a4621a3f9e6d", "title"=>"Referee Comments: Referee 1", "body"=>"\nReviewer 1's Review\n\nThe paper is nice, in my humble opinion. My compliments to you. I have written some little advice that might prove relevant to improve, after very minor revision, what you have done.\n\nThe presence of supplementary mouths or bifurcated tentacles can be normal (as it happens in Gastroblasta medusae, with multiple mouths, or in Cladonema, with bifurcated tentacles in the medusa, or in Cladocoryne, with bifurcated tentacles in the polyp). In the paper on Codonorchis I naively suggested that the multiple manubria that were observed in the senescent medusa might be due to the activation of genes leading to clonal morphs in the polyp stage. The \"do it again\" genes that make up the colony might have entered into action in the medusa, when some silencing of them became not so efficient. This passage of information from the polyp to the medusa, with mosaics of morphs, might be conducive to the formation of new bauplans, as I suggested also in the paper on the cnidarian premises of metazoan evolution and in other papers I published on the same subject. Mine were just wild speculations, you have the molecular evidence for them. After all, however, I imagined (without having any proof, besides comparative analyses) that such things might happen. However, if you think that this is squeezing my papers too much, you are free to ignore them.\n\nN.B. These are the general comments made by the reviewer when reviewing this paper. Specific points addressed during revision of the paper are not shown.", "isRemoved"=>false, "created"=>"2007-08-09T22:42:59Z", "lastModified"=>"2007-08-09T22:42:59Z", "creator"=>{"userId"=>"80539"}, "highlightedText"=>"", "competingInterestStatement"=>{"creatorWasPrompted"=>false, "hasCompetingInterests"=>false}, "parentArticle"=>{"doi"=>"info:doi/10.1371/journal.pone.0000694", "state"=>"published", "journals"=>{"PLoSONE"=>{"journalKey"=>"PLoSONE", "eIssn"=>"1932-6203", "title"=>"PLOS ONE"}}}, "replyTreeSize"=>0, "mostRecentActivity"=>"2007-08-09T22:42:59Z", "replies"=>[]}
• {"type"=>"COMMENT", "annotationUri"=>"info:doi/10.1371/annotation/e1b498c4-309f-48f1-b6ab-9f390746e4c4", "title"=>"Referee Comments: Referee 2", "body"=>"\nReviewer 2's Review\n\nThe authors report detailed and well-controlled gene silencing studies of Hox or Hox-like genes in a cnidarian hydrozoan, Eleutheria dichotoma. While there continues to be considerable debate and disagreement about the evolution of these genes in cnidarians and the relationship of these cnidarian genes to bilaterian genes, to date no functional assays have been possible. Without such assays, there has been considerable speculation, but little data. Now in an exhaustive series of experiments, the authors provide such data. As the authors write (p. 2-3): \"Unfortunately these [previous] studies have led to controversial results and functional studies are urgently needed to resolve controversies and learn about the function of Hox-like genes in early metazoan evolution.\" Thus this is a fascinating and important study with broad implications ranging from the methodological (functional assays in marine animals) to the evolutionary (origin of Hox gene function\n\nN.B. These are the general comments made by the reviewer when reviewing this paper. Specific points addressed during revision of the paper are not shown.", "isRemoved"=>false, "created"=>"2007-08-09T22:56:03Z", "lastModified"=>"2007-08-09T22:56:03Z", "creator"=>{"userId"=>"80539"}, "highlightedText"=>"", "competingInterestStatement"=>{"creatorWasPrompted"=>false, "hasCompetingInterests"=>false}, "parentArticle"=>{"doi"=>"info:doi/10.1371/journal.pone.0000694", "state"=>"published", "journals"=>{"PLoSONE"=>{"journalKey"=>"PLoSONE", "eIssn"=>"1932-6203", "title"=>"PLOS ONE"}}}, "replyTreeSize"=>0, "mostRecentActivity"=>"2007-08-09T22:56:03Z", "replies"=>[]}

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/947117"], "description"=>"<p>The five Hox-like genes, <i>Cnox-</i>1 to <i>Cnox-</i>5, display differential spatio-temporal expression patterns along the oral-aboral body axis. <i>Cnox-</i>1 (A₁–A₄) is expressed ectodermally in the so-called “Nesselring”, an area of undifferentiated cells lining the ring canal of medusae (cross section: A₃, A₄). <i>Cnox-</i>2 is expressed in the entoderm of developing medusa buds (B₁, B₂), in 2-day-old planula larvae ectodermally and orally (B₅), in 5-day-old planula larvae along the body column ectodermally (B₇) and at the oral pole of primary (B₉) and adult polyps (B₃; single polyp with 4 tentacles). <i>Cnox-</i>4 is exclusively expressed at the aboral pole of primary polyps (C₂). <i>Cnox-</i>3 expression perfectly marks the most ectodermal oral part of the manubrium (D₁, D₂). <i>Cnox-</i>5 shows a remarkable pattern of expression moving from aboral only in the planula larva (E₂) to both oral and aboral simultaneously in the metamorphosing polyp (E₄). NBT/X-phosphate (A₁, A₄, B₁–B₃, D₁) and fluorescein-labeled probes (A₂, B₅, B₇, B₉, C₂, D₂, E₂, E₄). Signals in B₅, B₇, B₉, C₂, E₂ and E₄ are overlaid with DAPI staining. Morphologies are shown in light microscopy (B₄, B₆, B₈, C₁, E₁, E₃). Scale bar is 50µm. A₂, D₁, E₁–E₄ are reprinted with permission from Elsevier Publishers <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000694#pone.0000694-Kamm1\" target=\"_blank\">[11]</a>.</p>", "links"=>[], "tags"=>["situ", "hox-like", "genes", "hydrozoan"], "article_id"=>617467, "categories"=>["Evolutionary Biology"], "users"=>["Wolfgang Jakob", "Bernd Schierwater"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000694.g003", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_situ_expression_of_Hox_like_genes_in_the_hydrozoan_Eleutheria_dichotoma_/617467", "title"=>"In situ expression of Hox-like genes in the hydrozoan <i>Eleutheria dichotoma.</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-08-01 02:04:27"}
• {"files"=>["https://ndownloader.figshare.com/files/947195"], "description"=>"<p>Putative homology assignment of Cnox genes according to Gauchat et al. (2000) <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000694#pone.0000694-Gauchat1\" target=\"_blank\">[43]</a>, Finnerty et al. (2003) <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000694#pone.0000694-Finnerty2\" target=\"_blank\">[21]</a> and Kuhn et al. (1996) <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000694#pone.0000694-Kuhn1\" target=\"_blank\">[18]</a>.</p>", "links"=>[], "tags"=>["nomenclature", "genes"], "article_id"=>617548, "categories"=>["Evolutionary Biology"], "users"=>["Wolfgang Jakob", "Bernd Schierwater"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000694.t002", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Homology_nomenclature_and_expression_of_Hox_ParaHox_like_genes_in_Cnidaria_/617548", "title"=>"Homology nomenclature and expression of Hox/ParaHox-like genes in Cnidaria.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2007-08-01 02:05:48"}
• {"files"=>["https://ndownloader.figshare.com/files/467547", "https://ndownloader.figshare.com/files/467672", "https://ndownloader.figshare.com/files/467737", "https://ndownloader.figshare.com/files/467807", "https://ndownloader.figshare.com/files/467875", "https://ndownloader.figshare.com/files/467939"], "description"=>"<div><p>Regulatory genes of the Antp class have been a major factor for the invention and radiation of animal bauplans. One of the most diverse animal phyla are the Cnidaria, which are close to the root of metazoan life and which often appear in two distinct generations and a remarkable variety of body forms. Hox-like genes have been known to be involved in axial patterning in the Cnidaria and have been suspected to play roles in the genetic control of many of the observed bauplan changes. Unfortunately RNAi mediated gene silencing studies have not been satisfactory for marine invertebrate organisms thus far. No direct evidence supporting Hox-like gene induced bauplan changes in cnidarians have been documented as of yet. Herein, we report a protocol for RNAi transfection of marine invertebrates and demonstrate that knock downs of Hox-like genes in Cnidaria create substantial bauplan alterations, including the formation of multiple oral poles (“heads”) by <em>Cnox-</em>2 and <em>Cnox-</em>3 inhibition, deformation of the main body axis by <em>Cnox</em>-5 inhibition and duplication of tentacles by <em>Cnox-</em>1 inhibition. All phenotypes observed in the course of the RNAi studies were identical to those obtained by morpholino antisense oligo experiments and are reminiscent of macroevolutionary bauplan changes. The reported protocol will allow routine RNAi studies in marine invertebrates to be established.</p></div>", "links"=>[], "tags"=>["hydrozoan", "bauplans", "hox-like", "genes"], "article_id"=>152186, "categories"=>["Evolutionary Biology"], "users"=>["Wolfgang Jakob", "Bernd Schierwater"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0000694.s001", "https://dx.doi.org/10.1371/journal.pone.0000694.s002", "https://dx.doi.org/10.1371/journal.pone.0000694.s003", "https://dx.doi.org/10.1371/journal.pone.0000694.s004", "https://dx.doi.org/10.1371/journal.pone.0000694.s005", "https://dx.doi.org/10.1371/journal.pone.0000694.s006"], "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Changing_Hydrozoan_Bauplans_by_Silencing_Hox_Like_Genes/152186", "title"=>"Changing Hydrozoan Bauplans by Silencing Hox-Like Genes", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2007-08-01 00:36:26"}
• {"files"=>["https://ndownloader.figshare.com/files/947046"], "description"=>"<p>Both morpholino oligo and RNAi experiments result in an increase of phenotypically abnormal medusae (PAMs) over time. Shown is the time course of increasing numbers of PAMs as a result of inhibition by double stranded RNA and inhibition by morpholino-antisense oligonucleotides. In each case control 1 is untreated animals cultured in normal seawater, and control 2 is a sense morpholino oligonucleotide or dsRNA targeted to the <i>Trox</i>-2 gene from the placozoan <i>Trichoplax adhaerens</i> respectively. Independent controls were run for each single experiment.</p>", "links"=>[], "tags"=>["phenotypic", "cnox-5"], "article_id"=>617397, "categories"=>["Evolutionary Biology"], "users"=>["Wolfgang Jakob", "Bernd Schierwater"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000694.g002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Time_course_for_the_development_of_phenotypic_effects_resulting_from_Cnox_5_gene_knock_down_/617397", "title"=>"Time course for the development of phenotypic effects resulting from Cnox-5 gene knock down.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-08-01 02:03:17"}
• {"files"=>["https://ndownloader.figshare.com/files/947217"], "description"=>"<p>Explanation is given in the text.</p>", "links"=>[], "tags"=>["phenotypes", "hydrozoan"], "article_id"=>617570, "categories"=>["Evolutionary Biology"], "users"=>["Wolfgang Jakob", "Bernd Schierwater"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000694.t001", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cnox_gene_expression_and_knock_down_phenotypes_in_the_hydrozoan_Eleutheria_dichotoma_/617570", "title"=>"Cnox gene expression and knock down phenotypes in the hydrozoan <i>Eleutheria dichotoma</i>.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2007-08-01 02:06:10"}
• {"files"=>["https://ndownloader.figshare.com/files/946950"], "description"=>"<p>Four of the five Hox-like genes produced phenotypically abnormal medusae (PAMs) in gene knock down studies. The main features of bauplan change relate to additional tentacle bifurcation (<i>Cnox</i>-1; see arrows), multiple oral poles or heads (<i>Cnox</i>-2), head duplication (<i>Cnox</i>-3; see arrows), and oral-aboral body axis deformation (<i>Cnox</i>-5). The two upper panels show life pictures of medusae transfected with double stranded RNA (B_1,2, C_1,2, D_1,2, E_1,2), and the lower panel medusae transfected with morpholino oligos (B_3,4, C_3,4, D_3,4, E_3,4). A_1,2 and A_3,4 are controls, scale bar is 100 µm. While we here unambiguously show that Hox-like genes can be silenced we think it would be premature to derive final conclusions on their functions yet.</p>", "links"=>[], "tags"=>["phenotypic", "experiments", "hox-like", "genes", "hydrozoan"], "article_id"=>617296, "categories"=>["Evolutionary Biology"], "users"=>["Wolfgang Jakob", "Bernd Schierwater"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000694.g001", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Observed_phenotypic_changes_in_knock_down_experiments_of_Hox_like_genes_in_the_hydrozoan_Eleutheria_dichotoma_/617296", "title"=>"Observed phenotypic changes in knock down experiments of Hox-like genes in the hydrozoan <i>Eleutheria dichotoma.</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-08-01 02:01:36"}

PMC Usage Stats | Further Information

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Relative Metric

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F1000Prime