Microarray and RNAi Analysis of P450s in Anopheles gambiae Male and Female Steroidogenic Tissues: CYP307A1 Is Required for Ecdysteroid Synthesis
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{"title"=>"Microarray and RNAi analysis of P450s in Anopheles gambiae male and female steroidogenic tissues: CYP307A1 is required for ecdysteroid synthesis", "type"=>"journal", "authors"=>[{"first_name"=>"Emilie", "last_name"=>"Pondeville", "scopus_author_id"=>"8988587200"}, {"first_name"=>"Jean Philippe", "last_name"=>"David", "scopus_author_id"=>"7401663936"}, {"first_name"=>"Emilie", "last_name"=>"Guittard", "scopus_author_id"=>"35344709400"}, {"first_name"=>"Annick", "last_name"=>"Maria", "scopus_author_id"=>"8588333900"}, {"first_name"=>"Jean Claude", "last_name"=>"Jacques", "scopus_author_id"=>"12791359200"}, {"first_name"=>"Hilary", "last_name"=>"Ranson", "scopus_author_id"=>"6603762935"}, {"first_name"=>"Catherine", "last_name"=>"Bourgouin", "scopus_author_id"=>"6603717350"}, {"first_name"=>"Chantal", "last_name"=>"Dauphin-Villemant", "scopus_author_id"=>"6701455373"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"372086521", "sgr"=>"84891898926", "pmid"=>"24324583", "scopus"=>"2-s2.0-84891898926", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0079861", "issn"=>"19326203"}, "id"=>"3db49687-787f-398b-b1b9-85ab341d30a2", "abstract"=>"In insects, the steroid hormone 20-hydroxyecdysone (20E) coordinates major developmental transitions. While the first and the final steps of 20E biosynthesis are characterized, the pathway from 7-dehydrocholesterol to 5β-ketodiol, commonly referred as the \"black box\", remains hypothetical and whether there are still unidentified enzymes is unknown. The black box would include some oxidative steps, which are believed to be mediated by P450 enzymes. To identify new enzyme(s) involved in steroid synthesis, we analyzed by small-scale microarray the expression of all the genes encoding P450 enzymes of the malaria mosquito Anopheles gambiae in active steroidogenic organs of adults, ovaries from blood-fed females and male reproductive tracts, compared to inactive steroidogenic organs, ovaries from non-blood-fed females. Some genes encoding P450 enzymes were specifically overexpressed in female ovaries after a blood-meal or in male reproductive tracts but only three genes were found to be overexpressed in active steroidogenic organs of both females and males: cyp307a1, cyp4g16 and cyp6n1. Among these genes, only cyp307a1 has an expression pattern similar to other mosquito steroidogenic genes. Moreover, loss-of-function by transient RNAi targeting cyp307a1 disrupted ecdysteroid production demonstrating that this gene is required for ecdysteroid biosynthesis in Anopheles gambiae.", "link"=>"http://www.mendeley.com/research/microarray-rnai-analysis-p450s-anopheles-gambiae-male-female-steroidogenic-tissues-cyp307a1-required", "reader_count"=>24, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>2, "Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>2, "Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Environmental Science"=>2, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>16, "Medicine and Dentistry"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>16}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>1}, "Environmental Science"=>{"Environmental Science"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United Kingdom"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1302898"], "description"=>"<p>From cholesterol to 20-hydroxyecdysone, the active steroid hormone. Characterized steps: <i>Nvd, neverland</i>, Rieske-domain oxygenase; <i>Phm, Phantom</i>, CYP306A1, 25-hydroxylase; <i>Dib, Disembodied</i>, CYP302A1, 22-hydroxylase; <i>Sad, Shadow</i>, CYP315A1, 2-hydroxylase; <i>Shd, Shade</i>, CYP314A1, 20-hydroxylase. Chemical modifications are shown in red on molecules. Putative steps of the “black box”, from 7-dehydro-cholesterol to 5ß-ketodiol, are represented within the dark grey box. [O] indicates oxidative step that might be catalyzed by a CYP enzyme. Some steps could be catalyzed by shroud, spook (CYP307A1) or spookier (CYP307A2). 5ßR: 5ß-reductase. 3ßHSD: 3ß-hydroxysteroid-dehydrogenase. Modified from Lafont <i>et al. </i><a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079861#pone.0079861-Lafont1\" target=\"_blank\">[4]</a>.</p>", "links"=>[], "tags"=>["pathway"], "article_id"=>868080, "categories"=>["Biological Sciences"], "users"=>["Emilie Pondeville", "Jean-Philippe David", "Emilie Guittard", "Annick Maria", "Jean-Claude Jacques", "Hilary Ranson", "Catherine Bourgouin", "Chantal Dauphin-Villemant"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0079861.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Biosynthetic_pathway_of_ecdysteroids_/868080", "title"=>"Biosynthetic pathway of ecdysteroids.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-04 03:15:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1302900"], "description"=>"<p>(A) RT-PCR analysis of <i>cyp4g16, cyp6n1</i> and <i>cyp307a1</i> expression pattern in males (T, testes; AG, accessory glands; G, gut and Malpighian tubules; C, carcass). <i>rpL17A</i> is used as a control gene. (B to F) <i>In situ</i> expression pattern of <i>cyp307a1</i>, <i>cyp4g16</i>, <i>cyp6n1</i> in males (T, testes; AG, accessory glands). (B) <i>cyp307a1</i> is detected in the anterior part of accessory glands. (C) <i>cyp4g16</i> is detected at the bottom of testes and in the posterior part of accessory glands. (D) <i>cyp4g16</i> is detected in the anterior and posterior gut. (E) <i>cyp6n1</i> is detected at the top of testes. (F) <i>cyp6n1</i> is expressed in the posterior gut and malpighian tubules. Black arrowheads show expression zones.</p>", "links"=>[], "tags"=>[], "article_id"=>868082, "categories"=>["Biological Sciences"], "users"=>["Emilie Pondeville", "Jean-Philippe David", "Emilie Guittard", "Annick Maria", "Jean-Claude Jacques", "Hilary Ranson", "Catherine Bourgouin", "Chantal Dauphin-Villemant"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0079861.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_pattern_of_cyp4g16_cyp6n1_and_cyp307a1_in_adult_males_/868082", "title"=>"Expression pattern of <i>cyp4g16</i>, <i>cyp6n1</i> and <i>cyp307a1</i> in adult males.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-04 03:15:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1302904"], "description"=>"<p>(A) RT-PCR analysis of <i>cyp314aA1</i> in ovaries of ds<i>gfp</i> and ds<i>cyp314a1</i> females 22h after blood-feeding. (B) RT-PCR analysis of <i>cyp307a1</i> in ovaries of ds<i>gfp</i> and ds<i>cyp307aA1</i> females 22h after blood-feeding. (C) <i>In vitro</i> ecdysteroid secretion of ovaries from ds<i>gfp</i> and ds<i>cyp314a1</i> females 22 h after blood-feeding. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079861#s2\" target=\"_blank\">Results</a> are expressed as mean ± SEM in 20E equivalents (in pg) per ovaries pair. (D) <i>In vitro</i> ecdysteroid secretion of ovaries from ds<i>gfp</i> and ds<i>cyp307a1</i> females 22 h after blood-feeding. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079861#s2\" target=\"_blank\">Results</a> are expressed as mean ± SEM in E equivalents (in pg) per ovaries pair. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079861#s2\" target=\"_blank\">Results</a> were subjected to statistical analysis using Mann-Whitney test (<b>**</b>, <i>P</i><0.01).</p>", "links"=>[], "tags"=>["ecdysteroid", "secretion", "ovaries", "dsrna", "injected"], "article_id"=>868086, "categories"=>["Biological Sciences"], "users"=>["Emilie Pondeville", "Jean-Philippe David", "Emilie Guittard", "Annick Maria", "Jean-Claude Jacques", "Hilary Ranson", "Catherine Bourgouin", "Chantal Dauphin-Villemant"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0079861.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_ecdysteroid_secretion_by_ovaries_of_dsRNA_injected_females_/868086", "title"=>"<i>In vitro</i> ecdysteroid secretion by ovaries of dsRNA injected females.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-04 03:15:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1302905"], "description"=>"<p>Expression ratios and p values (italic) for the genes encoding CYPs previously characterized as steroidogenic CYPs, in ovaries of blood-fed females at 5 h, 16 h, 22 h post blood-meal (PBM) and in male reproductive tracts (MRTs) compared to ovaries from non blood-fed females. CYP genes are listed according to their position in the 20E biosynthesis pathway. Genes showing a significant over transcription (ratio >1.5 and P<0.05) are shown in bold.</p>", "links"=>[], "tags"=>["genes", "encoding", "steroidogenic", "cyp", "ovaries", "mrts", "compared", "non"], "article_id"=>868087, "categories"=>["Biological Sciences"], "users"=>["Emilie Pondeville", "Jean-Philippe David", "Emilie Guittard", "Annick Maria", "Jean-Claude Jacques", "Hilary Ranson", "Catherine Bourgouin", "Chantal Dauphin-Villemant"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0079861.t001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_genes_encoding_steroidogenic_CYP_in_steroidogenic_ovaries_and_MRTs_compared_to_non_steroidogenic_ovaries_/868087", "title"=>"Expression of genes encoding steroidogenic CYP in steroidogenic ovaries and MRTs compared to non steroidogenic ovaries.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-12-04 03:15:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1302906"], "description"=>"<p>Only CYP genes overexpressed in ovaries of blood-fed females at least in one time point and/or in MRTs compared to ovaries from NBF females are listed. Values in bold indicate a ratio >1.5 and a p value<0.05. ND: Not detected or detected in less than 2 arrays. CYP genes previously characterized as involved in steroidogenesis are underlined. Candidate genes, overexpressed in ovaries of BF females, at least in one time point, and in MRTs compared to ovaries from NBF females, are in bold. PBM: post-blood meal. MRTs: Male reproductive tracts.</p>", "links"=>[], "tags"=>["genes", "transcribed", "steroidogenic", "ovaries", "mrts", "compared", "non"], "article_id"=>868088, "categories"=>["Biological Sciences"], "users"=>["Emilie Pondeville", "Jean-Philippe David", "Emilie Guittard", "Annick Maria", "Jean-Claude Jacques", "Hilary Ranson", "Catherine Bourgouin", "Chantal Dauphin-Villemant"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0079861.t002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CYP_genes_over_transcribed_in_steroidogenic_ovaries_and_or_in_steroidogenic_MRTs_compared_to_non_steroidogenic_ovaries_/868088", "title"=>"CYP genes over transcribed in steroidogenic ovaries and/or in steroidogenic MRTs compared to non steroidogenic ovaries.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-12-04 03:15:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1302907"], "description"=>"<div><p>In insects, the steroid hormone 20-hydroxyecdysone (20E) coordinates major developmental transitions. While the first and the final steps of 20E biosynthesis are characterized, the pathway from 7-dehydrocholesterol to 5β-ketodiol, commonly referred as the “black box”, remains hypothetical and whether there are still unidentified enzymes is unknown. The black box would include some oxidative steps, which are believed to be mediated by P450 enzymes. To identify new enzyme(s) involved in steroid synthesis, we analyzed by small-scale microarray the expression of all the genes encoding P450 enzymes of the malaria mosquito <i>Anopheles gambiae</i> in active steroidogenic organs of adults, ovaries from blood-fed females and male reproductive tracts, compared to inactive steroidogenic organs, ovaries from non-blood-fed females. Some genes encoding P450 enzymes were specifically overexpressed in female ovaries after a blood-meal or in male reproductive tracts but only three genes were found to be overexpressed in active steroidogenic organs of both females and males: <i>cyp307a1</i>, <i>cyp4g16</i> and <i>cyp6n1</i>. Among these genes, only <i>cyp307a1</i> has an expression pattern similar to other mosquito steroidogenic genes. Moreover, loss-of-function by transient RNAi targeting <i>cyp307a1</i> disrupted ecdysteroid production demonstrating that this gene is required for ecdysteroid biosynthesis in <i>Anopheles gambiae</i>.</p></div>", "links"=>[], "tags"=>["microarray", "rnai", "p450s", "steroidogenic", "ecdysteroid", "synthesis"], "article_id"=>868089, "categories"=>["Biological Sciences"], "users"=>["Emilie Pondeville", "Jean-Philippe David", "Emilie Guittard", "Annick Maria", "Jean-Claude Jacques", "Hilary Ranson", "Catherine Bourgouin", "Chantal Dauphin-Villemant"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0079861"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Microarray_and_RNAi_Analysis_of_P450s_in_Anopheles_gambiae_Male_and_Female_Steroidogenic_Tissues_CYP307A1_Is_Required_for_Ecdysteroid_Synthesis/868089", "title"=>"Microarray and RNAi Analysis of P450s in <i>Anopheles gambiae</i> Male and Female Steroidogenic Tissues: <i>CYP307A1</i> Is Required for Ecdysteroid Synthesis", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-12-04 03:15:32"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"2"}
  • {"unique-ip"=>"6", "full-text"=>"8", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"3"}
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Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences/Biochemistry", "average_usage"=>[266, 468, 593, 703, 804, 903, 993, 1084, 1171, 1256, 1339, 1422, 1492]}, {"subject_area"=>"/Medicine and health sciences/Epidemiology", "average_usage"=>[263, 452, 568, 671, 758, 847, 921, 1023, 1101, 1187, 1264, 1340, 1394]}, {"subject_area"=>"/Physical sciences", "average_usage"=>[254, 431, 547, 651, 748, 842, 932, 1017, 1098, 1178, 1259, 1336, 1404]}, {"subject_area"=>"/Physical sciences/Chemistry", "average_usage"=>[247, 429, 544, 647, 747, 842, 929, 1012, 1099, 1179, 1263, 1339, 1409]}]}
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