A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening
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{"title"=>"A High-Throughput Fluorescence-Based Assay System for Appetite-Regulating Gene and Drug Screening", "type"=>"journal", "authors"=>[{"first_name"=>"Yasuhito", "last_name"=>"Shimada", "scopus_author_id"=>"8054274700"}, {"first_name"=>"Minoru", "last_name"=>"Hirano", "scopus_author_id"=>"35316092100"}, {"first_name"=>"Yuhei", "last_name"=>"Nishimura", "scopus_author_id"=>"7403348248"}, {"first_name"=>"Toshio", "last_name"=>"Tanaka", "scopus_author_id"=>"56331841300"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"23300705", "doi"=>"10.1371/journal.pone.0052549", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "scopus"=>"2-s2.0-84871555083", "issn"=>"19326203", "pui"=>"366343889", "sgr"=>"84871555083"}, "id"=>"ca9eeb5c-5fd8-3ca4-8f86-44e2c4842fe6", "abstract"=>"The increasing number of people suffering from metabolic syndrome and obesity is becoming a serious problem not only in developed countries, but also in developing countries. However, there are few agents currently approved for the treatment of obesity. Those that are available are mainly appetite suppressants and gastrointestinal fat blockers. We have developed a simple and rapid method for the measurement of the feeding volume of Danio rerio (zebrafish). This assay can be used to screen appetite suppressants and enhancers. In this study, zebrafish were fed viable paramecia that were fluorescently-labeled, and feeding volume was measured using a 96-well microplate reader. Gene expression analysis of brain-derived neurotrophic factor (bdnf), knockdown of appetite-regulating genes (neuropeptide Y, preproinsulin, melanocortin 4 receptor, agouti related protein, and cannabinoid receptor 1), and the administration of clinical appetite suppressants (fluoxetine, sibutramine, mazindol, phentermine, and rimonabant) revealed the similarity among mechanisms regulating appetite in zebrafish and mammals. In combination with behavioral analysis, we were able to evaluate adverse effects on locomotor activities from gene knockdown and chemical treatments. In conclusion, we have developed an assay that uses zebrafish, which can be applied to high-throughput screening and target gene discovery for appetite suppressants and enhancers.", "link"=>"http://www.mendeley.com/research/highthroughput-fluorescencebased-assay-system-appetiteregulating-gene-drug-screening", "reader_count"=>62, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>4, "Researcher"=>14, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>17, "Student > Master"=>10, "Other"=>4, "Student > Bachelor"=>4, "Lecturer"=>1, "Professor"=>5}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>4, "Researcher"=>14, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>17, "Student > Master"=>10, "Other"=>4, "Student > Bachelor"=>4, "Lecturer"=>1, "Professor"=>5}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Engineering"=>1, "Environmental Science"=>2, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>31, "Medicine and Dentistry"=>10, "Neuroscience"=>10, "Business, Management and Accounting"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>10}, "Neuroscience"=>{"Neuroscience"=>10}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>31}, "Business, Management and Accounting"=>{"Business, Management and Accounting"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>4}, "Environmental Science"=>{"Environmental Science"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Netherlands"=>1, "Japan"=>1, "Brazil"=>3}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/519247"], "description"=>"<p>Downstream genes of appetite regulation in young zebrafish brains were measured after treatment with fluoxetine (A–D) and sibutramine (E–H). (A and E) <i>npy</i>, (B and F) <i>slc6a4</i>, (C and G) <i>htr1aa</i>, and (D and H) <i>htr1ab</i>. All values are mean ± SEM, <i>n</i> = 4, *<i>P</i><0.05, **<i>P</i><0.01.</p>", "links"=>[], "tags"=>["analyses", "fluoxetine", "sibutramine"], "article_id"=>189740, "categories"=>["Molecular Biology", "Biological Sciences", "Neuroscience", "Genetics", "Pharmacology"], "users"=>["Yasuhito Shimada", "Minoru Hirano", "Yuhei Nishimura", "Toshio Tanaka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052549.g005", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Gene_expression_analyses_following_fluoxetine_and_sibutramine_treatment_/189740", "title"=>"Gene expression analyses following fluoxetine and sibutramine treatment.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 18:08:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/519300"], "description"=>"<p>Morpholino oligonucleotides used in the present study.</p>", "links"=>[], "tags"=>["oligonucleotides"], "article_id"=>189799, "categories"=>["Molecular Biology", "Biological Sciences", "Neuroscience", "Genetics", "Pharmacology"], "users"=>["Yasuhito Shimada", "Minoru Hirano", "Yuhei Nishimura", "Toshio Tanaka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052549.t001", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Morpholino_oligonucleotides_used_in_the_present_study_/189799", "title"=>"Morpholino oligonucleotides used in the present study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-19 18:09:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/518957"], "description"=>"<p>(A) Schematic representation of the feeding assay. (B) The actual numbers of paramecia correlated with optical density at 500 nm (OD<sub>500</sub>). (C) Correlation between the relative amount of paramecia (P) and fluorescent intensity. (D) Fluorescent intensities of ingested paramecia in zebrafish. The fluorescent intensities of paramecia in zebrafish abdomen increased independently to the numbers of introduced-paramecia. All values are mean ± SEM, <i>n</i> = 15. (E) Fluorescent intensities after free-feeding of 4-10-Di-ASP-labeled paramecia. Fluorescent intensities of ingested paramecia were at maximum levels 1–3 h after feeding. All values are mean ± SEM, <i>n</i> = 15, *<i>P</i><0.05, **<i>P</i><0.01 compared with 1 h after feeding.</p>", "links"=>[], "tags"=>["feeding", "assay", "4-di-10-asp", "labeled"], "article_id"=>189442, "categories"=>["Molecular Biology", "Biological Sciences", "Neuroscience", "Genetics", "Pharmacology"], "users"=>["Yasuhito Shimada", "Minoru Hirano", "Yuhei Nishimura", "Toshio Tanaka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052549.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Zebrafish_feeding_assay_using_4_Di_10_ASP_labeled_paramecia_/189442", "title"=>"Zebrafish feeding assay using 4-Di-10-ASP labeled paramecia.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 18:06:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/519043"], "description"=>"<p>(A) Analysis by qPCR of brain-derived neurotrophic factor mRNA (<i>bdnf</i>) during paramecia feeding, <i>n</i> = 5, *<i>P</i><0.05. (B and C) Western blots showing embryonic injection of neuropeptide Y (<i>npy</i>) MO (B) and preproinsulin a (<i>insa</i>) MO. (D) Embryonic injection of <i>npy</i> MO and <i>npy</i> mRNA. Orexigenic gene knockdown decreased feeding volume. (E) <i>insa</i> MO and <i>insa</i> mRNA, orexigenic gene knockdown increased feeding volume at 7 dpf. <i>n</i> = 15, *<i>P</i><0.05. All values are mean ± SEM. (F and G) Knockdown of <i>npy</i> (F) and <i>insa</i> (G) affected locomotor activities at 5 dpf. <i>n</i> = 16, *<i>P</i><0.05. All values are mean ± SEM. (H) Detection of gene knockdown for splicing MOs in 2% (w/v) agarose gels stained with ethidium bromide.</p>", "links"=>[], "tags"=>["knocking", "genes", "appetite"], "article_id"=>189533, "categories"=>["Molecular Biology", "Biological Sciences", "Neuroscience", "Genetics", "Pharmacology"], "users"=>["Yasuhito Shimada", "Minoru Hirano", "Yuhei Nishimura", "Toshio Tanaka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052549.g002", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Gene_expression_and_the_effect_of_knocking_down_genes_associated_with_appetite_regulation_/189533", "title"=>"Gene expression and the effect of knocking down genes associated with appetite regulation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 18:07:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/519189"], "description"=>"<p>The effects of appetite-regulating drug treatment on spontaneous swimming (15 min) were measured in young zebrafish. (A) Fluoxetine, (B) Sibutramine, (C) Mazindol, (D) Phentermine, and (E) Rimonabant. All values are mean ± SEM, <i>n</i> = 16, *<i>P</i><0.01.</p>", "links"=>[], "tags"=>["appetite", "suppressants", "locomotor"], "article_id"=>189682, "categories"=>["Molecular Biology", "Biological Sciences", "Neuroscience", "Genetics", "Pharmacology"], "users"=>["Yasuhito Shimada", "Minoru Hirano", "Yuhei Nishimura", "Toshio Tanaka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052549.g004", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_appetite_suppressants_on_locomotor_activities_/189682", "title"=>"Effect of appetite suppressants on locomotor activities.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 18:08:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/519341"], "description"=>"<p><i>n</i> = 16, *<i>P</i><0.05, **<i>P</i><0.01. All values are mean ± SEM. nd; not detected. Red color indicates increase and green color indicates decrease in each parameter.</p>", "links"=>[], "tags"=>["morpholinos", "intake", "locomotor"], "article_id"=>189836, "categories"=>["Molecular Biology", "Biological Sciences", "Neuroscience", "Genetics", "Pharmacology"], "users"=>["Yasuhito Shimada", "Minoru Hirano", "Yuhei Nishimura", "Toshio Tanaka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0052549.t003", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_morpholinos_on_food_intake_and_locomotor_activity_/189836", "title"=>"Effects of morpholinos on food intake and locomotor activity.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-19 18:09:13"}
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PMC Usage Stats | Further Information

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  • {"unique-ip"=>"17", "full-text"=>"25", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
  • {"unique-ip"=>"4", "full-text"=>"6", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
  • {"unique-ip"=>"14", "full-text"=>"19", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
  • {"unique-ip"=>"6", "full-text"=>"4", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"12", "full-text"=>"23", "pdf"=>"6", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"13", "full-text"=>"12", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"11", "full-text"=>"10", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}

Relative Metric

{"start_date"=>"2012-01-01T00:00:00Z", "end_date"=>"2012-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences/Biochemistry", "average_usage"=>[316, 541, 663, 766, 856, 950, 1041, 1128, 1218, 1302, 1382, 1456, 1526, 1593, 1657, 1729, 1796, 1862, 1930, 1999, 2065, 2132, 2202, 2261, 2319]}, {"subject_area"=>"/Biology and life sciences/Biomechanics", "average_usage"=>[307, 515, 633, 733, 821, 912, 1008, 1100, 1202, 1281, 1363, 1425, 1481, 1569, 1644, 1721, 1788, 1842, 1913, 1975, 2020, 2087, 2135, 2175, 2228]}, {"subject_area"=>"/Biology and life sciences/Developmental biology", "average_usage"=>[313, 562, 690, 796, 893, 986, 1079, 1173, 1257, 1349, 1429, 1506, 1586, 1661, 1730, 1803, 1876, 1945, 2017, 2091, 2163, 2225, 2294, 2362, 2430]}, {"subject_area"=>"/Biology and life sciences/Neuroscience", "average_usage"=>[325, 542, 658, 757, 836, 925, 1015, 1095, 1177, 1263, 1338, 1411, 1477, 1537, 1599, 1663, 1726, 1795, 1857, 1921, 1979, 2041, 2099, 2150, 2205]}, {"subject_area"=>"/Biology and life sciences/Physiology", "average_usage"=>[307, 536, 653, 753, 839, 926, 1017, 1103, 1189, 1268, 1348, 1425, 1492, 1557, 1620, 1686, 1759, 1825, 1891, 1950, 2014, 2079, 2141, 2200, 2255]}, {"subject_area"=>"/Medicine and health sciences", "average_usage"=>[312, 547, 668, 769, 861, 953, 1046, 1135, 1224, 1307, 1388, 1464, 1536, 1606, 1676, 1744, 1812, 1882, 1954, 2020, 2089, 2155, 2218, 2282, 2344]}, {"subject_area"=>"/Medicine and health sciences/Anatomy and physiology", "average_usage"=>[308, 539, 659, 759, 852, 941, 1028, 1113, 1197, 1275, 1353, 1423]}, {"subject_area"=>"/Medicine and health sciences/Endocrinology", "average_usage"=>[315, 561, 698, 824, 914, 1014, 1127, 1230, 1339, 1437, 1505, 1602, 1669, 1731, 1794, 1868, 1945, 2023, 2083, 2140, 2231, 2286, 2342, 2407, 2460]}, {"subject_area"=>"/Medicine and health sciences/Physiology", "average_usage"=>[308, 535, 653, 754, 847, 930, 1020, 1103, 1188, 1268, 1351, 1420, 1487, 1550, 1614, 1679, 1757, 1830, 1898, 1961, 2024, 2086, 2150, 2216, 2266]}]}
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Net::HTTPTooManyRequests

Source
Scopus
Time
2019-08-19 08:24:25 UTC
Target URL
https://api.elsevier.com/content/search/index:SCOPUS?query=DOI(10.1371%2Fjournal.pone.0052549)
Trace

/app/models/concerns/networkable.rb:21:in `get_result'
/app/models/source.rb:165:in `get_data'
/app/models/retrieval_status.rb:47:in `perform_get_data'
/app/jobs/source_job.rb:52:in `block (2 levels) in perform'
/app/jobs/source_job.rb:51:in `block in perform'
/app/jobs/source_job.rb:35:in `each'
/app/jobs/source_job.rb:35:in `perform'