Transcriptome Analysis of Aedes aegypti Transgenic Mosquitoes with Altered Immunity
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{"title"=>"Transcriptome analysis of aedes aegypti transgenic mosquitoes with altered immunity", "type"=>"journal", "authors"=>[{"first_name"=>"Zhen", "last_name"=>"Zou", "scopus_author_id"=>"8374362300"}, {"first_name"=>"Jayme", "last_name"=>"Souza-Neto", "scopus_author_id"=>"26026100600"}, {"first_name"=>"Zhiyong", "last_name"=>"Xi", "scopus_author_id"=>"9242827700"}, {"first_name"=>"Vladimir", "last_name"=>"Kokoza", "scopus_author_id"=>"6602854009"}, {"first_name"=>"Sang Woon", "last_name"=>"Shin", "scopus_author_id"=>"7403643862"}, {"first_name"=>"George", "last_name"=>"Dimopoulos", "scopus_author_id"=>"19337048900"}, {"first_name"=>"Alexander", "last_name"=>"Raikhel", "scopus_author_id"=>"7005059447"}], "year"=>2011, "source"=>"PLoS Pathogens", "identifiers"=>{"pui"=>"362967260", "sgr"=>"81755177694", "issn"=>"15537366", "pmid"=>"22114564", "scopus"=>"2-s2.0-81755177694", "doi"=>"10.1371/journal.ppat.1002394", "isbn"=>"1553-7374 (Electronic) 1553-7366 (Linking)"}, "id"=>"1e55f29b-4c6a-32fc-a829-00efd4166b04", "abstract"=>"The mosquito immune system is involved in pathogen-elicited defense responses. The NF-kappaB factors REL1 and REL2 are downstream transcription activators of Toll and IMD immune pathways, respectively. We have used genome-wide microarray analyses to characterize fat-body-specific gene transcript repertoires activated by either REL1 or REL2 in two transgenic strains of the mosquito Aedes aegypti. Vitellogenin gene promoter was used in each transgenic strain to ectopically express either REL1 (REL1+) or REL2 (REL2+) in a sex, tissue, and stage specific manner. There was a significant change in the transcript abundance of 297 (79 up- and 218 down-regulated) and 299 (123 up- and 176 down-regulated) genes in fat bodies of REL1+ and REL2+, respectively. Over half of the induced genes had predicted functions in immunity, and a large group of these was co-regulated by REL1 and REL2. By generating a hybrid transgenic strain, which ectopically expresses both REL1 and REL2, we have shown a synergistic action of these NF-kappaB factors in activating immune genes. The REL1+ immune transcriptome showed a significant overlap with that of cactus (RNAi)-depleted mosquitoes (50%). In contrast, the REL2+ -regulated transcriptome differed from the relatively small group of gene transcripts regulated by RNAi depletion of a putative inhibitor of the IMD pathway, caspar (35 up- and 140 down-regulated), suggesting that caspar contributes to regulation of a subset of IMD-pathway controlled genes. Infections of the wild type Ae. aegypti with Plasmodium gallinaceum elicited the transcription of a distinct subset of immune genes (76 up- and 25 down-regulated) relative to that observed in REL1+ and REL2+ mosquitoes. Considerable overlap was observed between the fat body transcriptome of Plasmodium-infected mosquitoes and that of mosquitoes with transiently depleted PIAS, an inhibitor of the JAK-STAT pathway. PIAS gene silencing reduced Plasmodium proliferation in Ae. aegypti, indicating the involvement of the JAK-STAT pathway in anti-Plasmodium defense in this infection model.", "link"=>"http://www.mendeley.com/research/transcriptome-analysis-aedes-aegypti-transgenic-mosquitoes-altered-immunity", "reader_count"=>111, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>2, "Researcher"=>28, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>38, "Student > Postgraduate"=>5, "Student > Master"=>12, "Other"=>4, "Student > Bachelor"=>8, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>4}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>2, "Researcher"=>28, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>38, "Student > Postgraduate"=>5, "Student > Master"=>12, "Other"=>4, "Student > Bachelor"=>8, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>4}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>12, "Agricultural and Biological Sciences"=>88, "Medicine and Dentistry"=>1, "Neuroscience"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Chemistry"=>1, "Social Sciences"=>3, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Social Sciences"=>{"Social Sciences"=>3}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>88}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>12}, "Unspecified"=>{"Unspecified"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Colombia"=>1, "United States"=>4, "Sri Lanka"=>1, "Brazil"=>1, "United Kingdom"=>2, "Chile"=>1, "France"=>1, "Portugal"=>1, "Germany"=>1, "India"=>1, "Indonesia"=>1}, "group_count"=>5}

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  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360622/Figure_S1.pdf", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360629/Figure_S2.pdf", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360640/Figure_S3.pdf", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360650/Figure_S4.pdf", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360659/Figure_S5.pdf", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360672/Table_S1.docx", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360681/Table_S2.docx", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360688/Table_S3.docx", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360697/Table_S4.docx", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360704/Table_S5.docx", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360718/Table_S6.docx", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360728/Table_S7.docx", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360739/Table_S8.docx", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360750/Table_S9.docx", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/360767/Table_S10.docx"], "description"=>"<div><p>The mosquito immune system is involved in pathogen-elicited defense responses. The NF-κB factors REL1 and REL2 are downstream transcription activators of Toll and IMD immune pathways, respectively. We have used genome-wide microarray analyses to characterize fat-body-specific gene transcript repertoires activated by either REL1 or REL2 in two transgenic strains of the mosquito <em>Aedes aegypti</em>. Vitellogenin gene promoter was used in each transgenic strain to ectopically express either REL1 (REL1+) or REL2 (REL2+) in a sex, tissue, and stage specific manner. There was a significant change in the transcript abundance of 297 (79 up- and 218 down-regulated) and 299 (123 up- and 176 down-regulated) genes in fat bodies of REL1+ and REL2+, respectively. Over half of the induced genes had predicted functions in immunity, and a large group of these was co-regulated by REL1 and REL2. By generating a hybrid transgenic strain, which ectopically expresses both REL1 and REL2, we have shown a synergistic action of these NF-κB factors in activating immune genes. The REL1+ immune transcriptome showed a significant overlap with that of <em>cactus</em> (RNAi)-depleted mosquitoes (50%). In contrast, the REL2+ -regulated transcriptome differed from the relatively small group of gene transcripts regulated by RNAi depletion of a putative inhibitor of the IMD pathway, caspar (35 up- and 140 down-regulated), suggesting that caspar contributes to regulation of a subset of IMD-pathway controlled genes. Infections of the wild type <em>Ae. aegypti</em> with <em>Plasmodium gallinaceum</em> elicited the transcription of a distinct subset of immune genes (76 up- and 25 down-regulated) relative to that observed in REL1+ and REL2+ mosquitoes. Considerable overlap was observed between the fat body transcriptome of <em>Plasmodium</em>-infected mosquitoes and that of mosquitoes with transiently depleted <em>PIAS</em>, an inhibitor of the JAK-STAT pathway. <em>PIAS</em> gene silencing reduced <em>Plasmodium</em> proliferation in <em>Ae. aegypti</em>, indicating the involvement of the JAK-STAT pathway in anti-<em>Plasmodium</em> defense in this infection model.</p> </div>", "links"=>[], "tags"=>["transcriptome", "transgenic", "mosquitoes", "altered", "immunity"], "article_id"=>131282, "categories"=>["Microbiology"], "users"=>["Zhen Zou", "Jayme Souza-Neto", "Zhiyong Xi", "Vladimir Kokoza", "Sang Woon Shin", "George Dimopoulos", "Alexander Raikhel"], "doi"=>["http://dx.doi.org/10.1371/journal.ppat.1002394"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/Transcriptome_Analysis_of_Aedes_aegypti_Transgenic_Mosquitoes_with_Altered_Immunity/131282", "title"=>"Transcriptome Analysis of <em>Aedes aegypti</em> Transgenic Mosquitoes with Altered Immunity", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-11-17 00:21:22"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/711039/Figure_1.tif"], "description"=>"<p><b>A</b>) Functional classification of the REL1+ and REL2+ regulated transcriptome. Functional group abbreviations are: IMM, immunity; R/S/M, redox, stress and mitochondrion; CSR, chemosensory reception; DIG, digestive; C/S, cytoskeletal and structural; PROT, proteolysis; TRP, transport; R/T/T, replication, transcription, and translation; MET, metabolism; DIV, diverse functions; UNK, unknown functions. <b>B</b>) Venn diagram representing unique and shared transcriptome regulation in REL1+ and REL2+ transgenic mosquitoes, and <i>cactus</i>- and <i>caspar</i>-depleted mosquitoes. The overlapping regions represent genes that are concomitantly regulated in two, three, or four experimental conditions at the level of transcript abundance. The direction of gene transcript changes is indicated by upward-pointing and downward-pointing arrows. Green, Brown, Red, and Blue colors represent <i>caspar</i>-depleted, <i>cactus</i>-depleted, REL1+, and REL2+ transgenic mosquitoes, respectively. <b>C</b>) Hierarchical cluster analysis of fat body gene transcripts that were significantly regulated in at least two of the four experimental conditions; REL1+, REL2+, <i>cactus</i>- and <i>caspar</i>-depleted mosquitoes.</p>", "links"=>[], "tags"=>["transcriptome", "genes", "transgenic"], "article_id"=>381396, "categories"=>["Microbiology"], "users"=>["Zhen Zou", "Jayme Souza-Neto", "Zhiyong Xi", "Vladimir Kokoza", "Sang Woon Shin", "George Dimopoulos", "Alexander Raikhel"], "doi"=>["http://dx.doi.org/10.1371/journal.ppat.1002394.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_Comparative_transcriptome_analysis_of_fat_body_genes_in_REL1_and_REL2_transgenic_mosquitoes_/381396", "title"=>"Comparative transcriptome analysis of fat body genes in REL1+ and REL2+ transgenic mosquitoes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-17 00:23:16"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/711157/Figure_2.tif"], "description"=>"<p><b>A</b>) Transgenic mosquitoes with over-expressed <i>REL1</i> or <i>REL2</i> were used to generate hybrid transgenic mosquitoes over-expressing both of these factors. Fat bodies of REL1+, REL2+, and RE1+/REL2+ mosquitoes were dissected 24 h PBM and analyzed by means of quantitative RT-PCR for selected immune genes. A. Defensin A; B. Defensin C; C. TEP 20; D. CLIPB39. UGAL was used as a control. <b>B</b>) Both <i>Vg-REL1</i> and <i>Vg-REL2</i> transgenes were expressed in the hybrid mosquitoes. Total RNA isolated from females 24 h PBM were analyzed using specific primers for vitellogenin that recognize exclusively hybrid transgene mRNA. RT-PCR analysis indicated that both <i>Vg-REL1</i> and <i>Vg-REL2</i> transgenes were expressed in the hybrid mosquitoes. The same RNA samples were tested using actin specific primers as controls.</p>", "links"=>[], "tags"=>["rel1", "rel2", "activating"], "article_id"=>381517, "categories"=>["Microbiology"], "users"=>["Zhen Zou", "Jayme Souza-Neto", "Zhiyong Xi", "Vladimir Kokoza", "Sang Woon Shin", "George Dimopoulos", "Alexander Raikhel"], "doi"=>["http://dx.doi.org/10.1371/journal.ppat.1002394.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_Synergistic_action_of_REL1_and_REL2_in_activating_immune_genes_/381517", "title"=>"Synergistic action of REL1 and REL2 in activating immune genes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-17 00:25:17"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/711227/Figure_3.tif"], "description"=>"<p><b>A</b>) Functional classification of the PgMG- and PgFB-regulated transcriptomes. Functional group abbreviations are: IMM, immunity; R/S/M, redox, stress and mitochondrion; CSR, chemosensory reception; DIG, digestive; C/S, cytoskeletal and structural; PROT, proteolysis; TRP, transport; R/T/T, replication, transcription, and translation; MET, metabolism; DIV, diverse functions; UNK, unknown functions. <b>B</b>) Venn diagram representing unique and shared transcript regulation in PgMG, PgFB, and <i>cactus</i>- and <i>caspar</i>-depleted mosquitoes. The overlapping regions represent genes that are concomitantly regulated in two, three, or four experimental conditions at the level of transcript abundance. The direction of gene transcript regulation is indicated by upward-pointing and downward-pointing arrows. Green, Brown, Red, and Blue colors represent <i>caspar</i>-depleted, <i>cactus</i>-depleted, <i>PIAS</i>-depleted, and <i>Plasmodium</i>-infected mosquitoes, respectively.</p>", "links"=>[], "tags"=>["midgut-", "body-", "responsive"], "article_id"=>381586, "categories"=>["Microbiology"], "users"=>["Zhen Zou", "Jayme Souza-Neto", "Zhiyong Xi", "Vladimir Kokoza", "Sang Woon Shin", "George Dimopoulos", "Alexander Raikhel"], "doi"=>["http://dx.doi.org/10.1371/journal.ppat.1002394.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_Comparative_analysis_P_gallinaceum_midgut_and_fat_body_responsive_transcriptomes_/381586", "title"=>"Comparative analysis <i>P. gallinaceum</i> midgut- and fat body- responsive transcriptomes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-17 00:26:26"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/711338/Figure_4.tif"], "description"=>"<p><b>A</b>) Venn diagram representing unique and shared gene transcript regulation in PgFB, REL1+, and REL2+ transgenic mosquitoes. The overlapping regions represent genes that are concomitantly regulated in two or three experimental conditions. The direction of gene regulation is represented by upward-pointing and downward-pointing arrows. <b>B</b>) Hierarchical cluster analysis of the genes that were regulated in REL1+, REL2+, <i>PIAS</i>-depleted, and <i>P. gallinaceum</i> challenged mosquitoes.</p>", "links"=>[], "tags"=>["responsive"], "article_id"=>381704, "categories"=>["Microbiology"], "users"=>["Zhen Zou", "Jayme Souza-Neto", "Zhiyong Xi", "Vladimir Kokoza", "Sang Woon Shin", "George Dimopoulos", "Alexander Raikhel"], "doi"=>["http://dx.doi.org/10.1371/journal.ppat.1002394.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_Comparative_analysis_of_the_P_gallinaceum_fat_body_responsive_to_the_REL1_and_REL2_regulated_transcriptomes_/381704", "title"=>"Comparative analysis of the <i>P. gallinaceum</i> fat body responsive to the <i>REL1</i>- and <i>REL2</i>-regulated transcriptomes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-17 00:28:24"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/711444/Figure_5.tif"], "description"=>"<p>The effect of cactus, caspar, and PIAS in the defense against avian malaria parasite was compared. Depletion of <i>PIAS</i> and <i>cactus</i> significantly decreased the number of survival oocysts. However, the RNAi knock-down of <i>caspar</i> had no effect in the infection of avian malaria parasite to mosquito <i>Ae. aegypti</i>. Prefix i indicates the RNAi-mediated depletion of certain genes by direct injection of their corresponding dsRNAs. iLuc (luciferase dsRNA) was used as controls. The number of fully developed oocysts in each midgut was shown as a circle. The mean number of parasite oocysts for each group was indicated by a black bar.</p>", "links"=>[], "tags"=>["pias", "avian", "malaria"], "article_id"=>381804, "categories"=>["Microbiology"], "users"=>["Zhen Zou", "Jayme Souza-Neto", "Zhiyong Xi", "Vladimir Kokoza", "Sang Woon Shin", "George Dimopoulos", "Alexander Raikhel"], "doi"=>["http://dx.doi.org/10.1371/journal.ppat.1002394.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_The_role_of_PIAS_in_the_defense_against_avian_malaria_parasite_/381804", "title"=>"The role of PIAS in the defense against avian malaria parasite.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-17 00:30:04"}

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