Light-Dependent Development of Circadian Gene Expression in Transgenic Zebrafish
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{"title"=>"Light-dependent development of circadian gene expression in transgenic zebrafish", "type"=>"journal", "authors"=>[{"first_name"=>"Maki", "last_name"=>"Kaneko", "scopus_author_id"=>"7403595666"}, {"first_name"=>"Gregory M.", "last_name"=>"Cahill", "scopus_author_id"=>"7005277631"}], "year"=>2005, "source"=>"PLoS Biology", "identifiers"=>{"pui"=>"40412651", "sgr"=>"15744376086", "issn"=>"15449173", "pmid"=>"15685291", "scopus"=>"2-s2.0-15744376086", "doi"=>"10.1371/journal.pbio.0030034", "isbn"=>"1545-7885 (Electronic)"}, "id"=>"ceef0ab8-8ed2-3621-b0b4-55a41dcb78a9", "abstract"=>"The roles of environmental stimuli in initiation and synchronization of circadian oscillation during development appear to vary among different rhythmic processes. In zebrafish, a variety of rhythms emerge in larvae only after exposure to light-dark (LD) cycles, whereas zebrafish period3 (per3) mRNA has been reported to be rhythmic from day 1 of development in constant conditions. We generated transgenic zebrafish in which expression of the firefly luciferase (luc) gene is driven by the zebrafish per3 promoter. Live larvae from these lines are rhythmically bioluminescent, providing the first vertebrate system for high-throughput measurement of circadian gene expression in vivo. Circadian rhythmicity in constant conditions was observed only after 5-6 d of development, and only if the fish were exposed to LD signals after day 4. Regardless of light exposure, a novel developmental profile was observed, with low expression during the first few days and a rapid increase when active swimming begins. Ambient temperature affected the developmental profile and overall levels of per3 and luc mRNA, as well as the critical days in which LD cycles were needed for robust bioluminescence rhythms. In summary, per3-luc zebrafish has revealed complex interactions among developmental events, light, and temperature in the expression of a clock gene.", "link"=>"http://www.mendeley.com/research/lightdependent-development-circadian-gene-expression-transgenic-zebrafish-9", "reader_count"=>68, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>8, "Researcher"=>16, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>13, "Student > Postgraduate"=>5, "Student > Master"=>6, "Other"=>3, "Student > Bachelor"=>6, "Lecturer > Senior Lecturer"=>1, "Professor"=>7}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>8, "Researcher"=>16, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>13, "Student > Postgraduate"=>5, "Student > Master"=>6, "Other"=>3, "Student > Bachelor"=>6, "Lecturer > Senior Lecturer"=>1, "Professor"=>7}, "reader_count_by_subject_area"=>{"Environmental Science"=>3, "Biochemistry, Genetics and Molecular Biology"=>6, "Agricultural and Biological Sciences"=>45, "Medicine and Dentistry"=>3, "Neuroscience"=>4, "Veterinary Science and Veterinary Medicine"=>1, "Psychology"=>3, "Immunology and Microbiology"=>1, "Sports and Recreations"=>1, "Unspecified"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Neuroscience"=>{"Neuroscience"=>4}, "Psychology"=>{"Psychology"=>3}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Sports and Recreations"=>{"Sports and Recreations"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>45}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}, "Environmental Science"=>{"Environmental Science"=>3}, "Unspecified"=>{"Unspecified"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"New Zealand"=>1, "Canada"=>1, "Sweden"=>1, "United States"=>1, "United Kingdom"=>2, "South Africa"=>1, "Chile"=>1, "India"=>2}, "group_count"=>5}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/962985"], "description"=>"<div><p>(A) A schematic diagram showing how embryos were entrained and collected for RNA extraction. Embryos were entrained in 14:10 LD cycles (lights on at 8 A.M.; lights off at 10 P.M. CST) at two different temperatures, 22 °C and 28.5 °C. On day 6, half of the animals were sacrificed for RNA at 10 A.M. (2 h after lights-on) and at 10 P.M. (at lights-off). The rest of the animals were transferred to DD at 22 °C at 10 P.M. on that day, and sacrificed for RNA on day 10 at 10 A.M. and 10 P.M. The white and black bars represent day and night, respectively, and the gray bars the time at which lights would have been on had the LD cycles continued. The arrowheads indicate the time at which the animals were sacrificed for RNA extraction.</p>\n <p>(B) Relative mRNA level per animal for <i>per3</i> on days 6 and 10 of the experiment quantified by real-time qPCR. The levels were normalized to the value of the 10 A.M. time point on day 6 at 28.5 °C.</p>\n <p>(C) Relative RNA level per animal for <i>luc</i> measured from the same samples used in (B). For both (B) and (C), averages of three experiments are shown. Error bars represent SEM.</p></div>", "links"=>[], "tags"=>["mrna", "elevated", "temperatures"], "article_id"=>632948, "categories"=>["Molecular Biology", "Neuroscience", "Genetics"], "users"=>["Maki Kaneko", "Gregory M Cahill"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0030034.g005", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Levels_of_per3_and_luc_mRNA_Are_Elevated_by_High_Temperatures_during_Development_/632948", "title"=>"Levels of <i>per3</i> and <i>luc</i> mRNA Are Elevated by High Temperatures during Development", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:08:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/473381"], "description"=>"<div><p>The roles of environmental stimuli in initiation and synchronization of circadian oscillation during development appear to vary among different rhythmic processes. In zebrafish, a variety of rhythms emerge in larvae only after exposure to light-dark (LD) cycles, whereas zebrafish <em>period3 (per3)</em> mRNA has been reported to be rhythmic from day 1 of development in constant conditions. We generated transgenic zebrafish in which expression of the firefly <em>luciferase (luc)</em> gene is driven by the zebrafish <em>per3</em> promoter. Live larvae from these lines are rhythmically bioluminescent, providing the first vertebrate system for high-throughput measurement of circadian gene expression in vivo. Circadian rhythmicity in constant conditions was observed only after 5–6 d of development, and only if the fish were exposed to LD signals after day 4. Regardless of light exposure, a novel developmental profile was observed, with low expression during the first few days and a rapid increase when active swimming begins. Ambient temperature affected the developmental profile and overall levels of <em>per3</em> and <em>luc</em> mRNA, as well as the critical days in which LD cycles were needed for robust bioluminescence rhythms. In summary, <em>per3-luc</em> zebrafish has revealed complex interactions among developmental events, light, and temperature in the expression of a clock gene.</p> </div>", "links"=>[], "tags"=>["light-dependent", "circadian", "transgenic", "zebrafish"], "article_id"=>153270, "categories"=>["Molecular Biology", "Neuroscience", "Genetics"], "users"=>["Maki Kaneko", "Gregory M Cahill"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0030034", "stats"=>{"downloads"=>5, "page_views"=>72, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Light_Dependent_Development_of_Circadian_Gene_Expression_in_Transgenic_Zebrafish/153270", "title"=>"Light-Dependent Development of Circadian Gene Expression in Transgenic Zebrafish", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-01-20 11:34:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/963075"], "description"=>"<div><p>(A) Average plot of bioluminescence rhythms in DD. Animals were entrained in 14:10 LD cycles for 6 d at 22 °C and tested for approximately 6.5 d in DD.</p>\n <p>(B) Representative plot of bioluminescence rhythm in DD for an individual.</p>\n <p>(C) Average plot of bioluminescence rhythms in 14:10 LD cycles. Animals were entrained in 6 LD cycles at 22 °C prior to the monitoring.</p>\n <p>(D) Individual plot of bioluminescence rhythm in LD cycles. For each of DD and LD experiments, two experiments have been performed with essentially the same results. Only one of two experiments is shown for each of DD and LD. The first 12 h of data were deleted from each plot. Black and white bars on top of each plot represent the time when lights were off and on, respectively. Numbers of animals that were averaged is given at top right corner of (A) and (C). Error bars represent SEM in (A) and (C).</p></div>", "links"=>[], "tags"=>["rhythms", "mediated", "dd", "ld"], "article_id"=>633031, "categories"=>["Molecular Biology", "Neuroscience", "Genetics"], "users"=>["Maki Kaneko", "Gregory M Cahill"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0030034.g006", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bioluminescence_Rhythms_Mediated_by_per3_luc_in_DD_and_LD_Measured_for_Six_Days_/633031", "title"=>"Bioluminescence Rhythms Mediated by <i>per3</i>-<i>luc</i> in DD and LD Measured for Six Days", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:08:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/962906"], "description"=>"<p>Transgenic embryos were entrained by two, four, or six LD cycles at either 22 °C or 28.5 °C, and monitored for bioluminescence in DD at 21–24 °C. (A) Two LDs at 22 °C, (B) two LDs at 28.5 °C, (C) four LDs at 22 °C, (D) four LDs at 28.5 °C, (E) six LDs at 22 °C, and (F) six LDs at 28.5 °C. The insets in (E) and (F) show the last 3 d of the record with magnified y-axis scales. Black and white bars on top of each plot represent the times when the lights were off and on, respectively. Actual amount of bioluminescence in cps is averaged and plotted in each graph. Number of animals that were averaged is given at top right corner of each plot. Error bars represent SEM. Results of one of two identical experiments with similar results are shown here.</p>", "links"=>[], "tags"=>["genetics and genomics", "molecular biology", "neuroscience"], "article_id"=>632873, "categories"=>["Molecular Biology", "Neuroscience", "Genetics"], "users"=>["Maki Kaneko", "Gregory M Cahill"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0030034.g004", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_Temperature_on_Development_of_per3_luc_Expression_/632873", "title"=>"Effects of Temperature on Development of <i>per3-luc</i> Expression", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:07:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/962606"], "description"=>"<p>The top graphic shows the exon-intron structures of <i>per3</i> and the flanking gene. The BAC clone 8M06 screened for the first coding exon of <i>per3</i> is approximately 72 kb long, and extends from about 26 kb upstream of exon 1 to intron 19 of <i>per3</i>. The bottom graphic shows the magnified view of the first coding exon in the BAC 8M06 and the modified BAC construct. The white and black boxes represent noncoding and coding sequences, respectively, of the first coding exon of <i>per3</i>. The coding sequence of this exon was replaced with an approximately 3-kb fragment containing <i>luc</i> and <i>Km<sup>r</sup></i>. Arrows under the construct represent primers used for the screening of transgenic lines.</p>", "links"=>[], "tags"=>["genetics and genomics", "molecular biology", "neuroscience"], "article_id"=>632586, "categories"=>["Molecular Biology", "Neuroscience", "Genetics"], "users"=>["Maki Kaneko", "Gregory M Cahill"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0030034.g001", "stats"=>{"downloads"=>3, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_Map_of_the_per3_luc_Construct_/632586", "title"=>"Schematic Map of the <i>per3-luc</i> Construct", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:05:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/962685"], "description"=>"<p>Embryos hemizygous for <i>per3-luc</i> were collected and monitored for bioluminescence while exposed to different numbers of 14:10 LD cycles starting on day 1 postfertilization followed by DD. (A) One LD, (B) one LD, (C) two LD, (D) three LD, (E) four LD, (F) five LD, (G) six LD, and (H) six LD. In (I), embryos were exposed to two LDs, one on day 1 and the other on day 6 of development. Black and white bars on top of each plot represent the times when the lights were off and on, respectively. Since overall bioluminescence levels can vary among clutches and experiments, normalized bioluminescence was averaged and plotted in each graph. Number of animals that were averaged is given at top right corner of each plot. Error bars represent standard error of the mean (SEM) For the one-LD and six-LD groups, plots for two experiments are shown here. These experiments showed small differences in developmental profiles, possibly due to differences in room temperature (about 1 °C), therefore could not be pooled. For the two- to five-LD groups, data from two experiments were pooled. The small but abrupt increase of luminescence that occurred only during the light period of LD cycles is considered an artifact made visible by low bioluminescence counts during the first several days of development, because the same level of fluctuation was observed in empty wells under LD condition.</p>", "links"=>[], "tags"=>["embryos", "numbers", "ld", "cycles"], "article_id"=>632663, "categories"=>["Molecular Biology", "Neuroscience", "Genetics"], "users"=>["Maki Kaneko", "Gregory M Cahill"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0030034.g002", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bioluminescence_in_Embryos_That_Experienced_Different_Numbers_of_LD_Cycles_during_Development_/632663", "title"=>"Bioluminescence in Embryos That Experienced Different Numbers of LD Cycles during Development", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/962757"], "description"=>"<div><p>Embryos hemizygous for <i>per3-luc</i> were collected from naturally breeding parents and kept in 14:10 LD cycles (lights on at 8 A.M. CST) at 22 °C for 6 d. Larval fish were shifted to DD at the end of the light phase on day 6. Total RNA was extracted from 2- to 3-d-old embryos and 7- to 8-d-old larval fish, and was subjected to real-time PCR for <i>per3</i> and <i>luc</i> mRNA levels.</p>\n <p>(A) Expression of <i>per3</i> mRNA per embryo on day 3 was determined every 6 h starting at 10 A.M. (2 h after lights-on).</p>\n <p>(B) The <i>per3</i> mRNA per animal on day 8 was measured every 4 h starting at 10 A.M.</p>\n <p>(C) Levels of <i>luc</i> mRNA on day 3 were determined as in (A). White and black bars at the bottom represent light and dark phases, respectively, for (A) and (C).</p>\n <p>(D) Cycling of <i>luc</i> mRNA on day 8 was determined as in (B). The gray and black bars represent the time when the light would have been on and off, respectively, had the LD cycles continued.</p>\n <p>For each of <i>per3</i> and <i>luc</i>, mRNA levels were normalized to the peak level on day 8 (10 A.M. time point). The y-axis scales were set at 120% maximum for all plots to allow direct comparison of mRNA extracted on days 3 and 8. The x-axis scales are given in both hours and days postfertilization to facilitate the comparison with <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0030034#pbio-0030034-g002\" target=\"_blank\">Figure 2</a>. In order to show more detailed temporal profiles of mRNAs on day 3, plots with smaller y-axis scales were shown in the insets at top right corners of (A) and (C). Each plot is the average of three identical experiments, and error bars represent SEM. An identical experiment was also done at 24 °C with essentially the same results (unpublished data).</p></div>", "links"=>[], "tags"=>["mrna"], "article_id"=>632735, "categories"=>["Molecular Biology", "Neuroscience", "Genetics"], "users"=>["Maki Kaneko", "Gregory M Cahill"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.0030034.g003", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Temporal_Expression_of_luc_mRNA_Is_Similar_to_That_of_per3_during_Development_/632735", "title"=>"Temporal Expression of <i>luc</i> mRNA Is Similar to That of <i>per3</i> during Development", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 09:06:43"}

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  • {"unique-ip"=>"4", "full-text"=>"4", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"12"}
  • {"unique-ip"=>"11", "full-text"=>"9", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"1"}
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  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"3"}
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  • {"unique-ip"=>"5", "full-text"=>"2", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"8", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2016", "month"=>"10"}
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  • {"unique-ip"=>"7", "full-text"=>"6", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"29", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"12"}
  • {"unique-ip"=>"7", "full-text"=>"2", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2017", "month"=>"1"}
  • {"unique-ip"=>"6", "full-text"=>"9", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"2"}
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  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"1"}
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  • {"unique-ip"=>"11", "full-text"=>"11", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"4"}
  • {"unique-ip"=>"6", "full-text"=>"7", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"6"}
  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"8"}
  • {"unique-ip"=>"8", "full-text"=>"7", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"7"}
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  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
  • {"unique-ip"=>"8", "full-text"=>"4", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"2", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
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  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"6", "full-text"=>"3", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"9", "full-text"=>"10", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"5", "full-text"=>"3", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"6", "full-text"=>"4", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
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Relative Metric

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