MicroRNA-3906 Regulates Fast Muscle Differentiation through Modulating the Target Gene homer-1b in Zebrafish Embryos
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{"title"=>"MicroRNA-3906 Regulates Fast Muscle Differentiation through Modulating the Target Gene homer-1b in Zebrafish Embryos", "type"=>"journal", "authors"=>[{"first_name"=>"Cheng Yung", "last_name"=>"Lin", "scopus_author_id"=>"36087714700"}, {"first_name"=>"Jie Shin", "last_name"=>"Chen", "scopus_author_id"=>"55813730600"}, {"first_name"=>"Moo Rung", "last_name"=>"Loo", "scopus_author_id"=>"56375409200"}, {"first_name"=>"Chung Ching", "last_name"=>"Hsiao", "scopus_author_id"=>"55814118000"}, {"first_name"=>"Wen Yen", "last_name"=>"Chang", "scopus_author_id"=>"55813868700"}, {"first_name"=>"Huai Jen", "last_name"=>"Tsai", "scopus_author_id"=>"7402649201"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"23936160", "sgr"=>"84881121282", "doi"=>"10.1371/journal.pone.0070187", "scopus"=>"2-s2.0-84881121282", "pui"=>"369507654", "isbn"=>"10.1371/journal.pone.0070187", "issn"=>"19326203"}, "id"=>"8a672aba-81ad-31de-98ca-348289e801e0", "abstract"=>"A microRNA, termed miR-In300 or miR-3906, suppresses the transcription of myf5 through silencing dickkopf-related protein 3 (dkk3r/dkk3a) during early development when myf5 is highly transcribed, but not at late stages when myf5 transcription is reduced. Moreover, after 24 hpf, when muscle cells are starting to differentiate, Dkk3a could not be detected in muscle tissue at 20 hpf. To explain these reversals, we collected embryos at 32 hpf, performed assays, and identified homer-1b, which regulates calcium release from sarcoplasmic reticulum, as the target gene of miR-3906. We further found that either miR-3906 knockdown or homer-1b overexpression increased expressions of fmhc4 and atp2a1 of calcium-dependent fast muscle fibrils, but not slow muscle fibrils, and caused a severe disruption of sarcomeric actin and Z-disc structure. Additionally, compared to control embryos, the intracellular calcium concentration ([Ca(2+)]i) of these treated embryos was increased as high as 83.9-97.3% in fast muscle. In contrast, either miR-3906 overexpression or homer-1b knockdown caused decreases of [Ca(2+)]i and, correspondingly, defective phenotypes in fast muscle. These defects could be rescued by inducing homer-1b expression at later stage. These results indicate that miR-3906 controls [Ca(2+)]i homeostasis in fast muscle through fine tuning homer-1b expression during differentiation to maintain normal muscle development.", "link"=>"http://www.mendeley.com/research/microrna3906-regulates-fast-muscle-differentiation-through-modulating-target-gene-homer1b-zebrafish", "reader_count"=>15, "reader_count_by_academic_status"=>{"Student > Doctoral Student"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>4, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>2, "Unspecified"=>1}, "reader_count_by_user_role"=>{"Student > Doctoral Student"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>4, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>2, "Unspecified"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>9, "Medicine and Dentistry"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>9}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}}, "reader_count_by_country"=>{"United Kingdom"=>1, "Portugal"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1138356"], "description"=>"<p>(<b>A</b>) The schematic diagrams of plasmid constructs having a 1.1 kb fragment upstream of <i>myf5</i> gene and having various deletions of exon 1 of <i>myf5</i> which are proximal upstream regulatory segments of <i>miR-3906.</i> (<b>B</b>) Luciferase activity assay in zebrafish embryos at 24 hpf. Plasmids phRL-exon1-1, phRL-exon1-2, phRL-exon1-3, phRL-exon1-4 and phRL-exon1-5 were co-injected individually with pGL3-TK, an internal control, into one-cell embryos. The <i>luc</i> expression of embryos injected with plasmid phRL-exon1-1 served as 1. The relative <i>luc</i> expression level of each combination was examined. (<b>C</b>) Luciferase activity assay in zebrafish embryos at 16, 24 and 32 hpf. Plasmids phRL-<i>myf5</i>-promoter and phRL-exon1-1 were co-injected individually with pGL3-TK, an internal control, into one-cell embryos. The <i>luc</i> expression of each plasmid at 16 hpf served as 1. The relative <i>luc</i> expression levels of phRL-<i>myf5</i>-promoter and phRL-exon1-1 at 24 and 32 hpf were examined. Data were presented as means±SD from three independent experiments (n = 3). Student's <i>t</i>-test was used to calculate the differences among data. *: indicates significant difference at <i>P<0.05</i>. **: indicates significant difference at <i>P<0.01</i>.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "proximal", "upstream", "controlling", "transcription"], "article_id"=>760763, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0070187.g008", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_proximal_upstream_cis_elements_of_miR_3906_are_capable_of_controlling_the_transcription_of_miR_3906_/760763", "title"=>"The proximal upstream <i>cis</i>-elements of <i>miR-3906</i> are capable of controlling the transcription of <i>miR-3906</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-31 01:50:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1138338"], "description"=>"<p>(<b>A</b>, <b>D</b>) Compared with WT, injection of <i>miR-3906</i>-MO increased the expression level of <i>homer-1b</i> mRNA in embryos at 24 hpf (<b>A </b><i>vs</i>. <b>B</b>) and 32 hpf (<b>D </b><i>vs</i>. <b>E</b>). Injection of <i>miR-3906</i> dsR (mature double-strand <i>miR-3906</i>) reduced the expression level of <i>homer-1b</i> mRNA in the front of trunk muscle and was absent in the tail region at 24 hpf (<b>A </b><i>vs</i>. <b>C</b>) and 32-hpf (<b>D </b><i>vs</i>. <b>F</b>). (<b>G</b>) The expression levels of <i>homer-1b</i> in WT embryos, <i>miR-3906</i>-MO-injected embryos and <i>miR-3906</i> dsR-injected embryos at 24 hpf and 32 hpf were quantified. Each one was carried out with 100 embryos, and triplicate experiments were performed (n = 3). The numbers shown in the lower-right corner of panels A–F indicate the number of phenotypes out of the total number of embryos examined. SigmaPlot software was used to perform Student’s <i>t</i>-test. *: indicates the difference at <i>P<0.05</i> level; **: indicates the difference at <i>P<0.01</i> level.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "affected", "mrna", "zebrafish"], "article_id"=>760745, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0070187.g003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Change_of_miR_3906_level_affected_the_amount_of_homer_1b_mRNA_in_zebrafish_embryos_/760745", "title"=>"Change of <i>miR-3906</i> level affected the amount of <i>homer-1b</i> mRNA in zebrafish embryos.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-31 01:50:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1138336"], "description"=>"<p>(<b>A, D, G</b>) Using whole-mount <i>in situ</i> hybridization to detect the temporal and spatial expression patterns of <i>myf5</i>, (<b>B, E, H</b>) <i>miR-3906</i> and (<b>C, F, I</b>) <i>homer-1b</i> in zebrafish embryos at 20 hpf, 24 hpf and 32 hpf. At 20 hpf, (<b>A</b>) <i>myf5</i> was expressed in the pre-somitic mesoderm and newly formed somites (indicated by { ), but it was absent in mature somites (indicated by [); (<b>B</b>) <i>miR-3906</i> and (<b>C</b>) <i>homer-1b</i> were detected in mature somites. At 24 hpf, (<b>D</b>) <i>myf5</i> was detected in the pre-somitic mesoderm, while (<b>E</b>) <i>miR-3906</i> and (<b>F</b>) <i>homer-1b</i> gradually increased expression in mature somites. At 32 hpf, (<b>G</b>) <i>myf5</i> was expressed in the edge region of trunk muscles (arrowhead), but (<b>H</b>) <i>miR-3906</i> and (<b>I</b>) <i>homer-1b</i> were expressed in all trunk muscles. Cross section (dotted lines) showed that (<b>F’</b>) <i>miR-3906</i> and (<b>I’</b>) <i>homer-1b</i> were expressed in fast muscle (arrows).</p>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "patterns", "somites", "zebrafish"], "article_id"=>760743, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0070187.g002", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_expression_patterns_of_myf5_miR_3906_and_homer_1b_in_the_trunk_somites_of_zebrafish_embryos_/760743", "title"=>"The expression patterns of <i>myf5</i>, <i>miR-3906</i> and <i>homer-1b</i> in the trunk somites of zebrafish embryos.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-31 01:50:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1138346"], "description"=>"<p>(<b>A–C</b>) Wild-type (WT) zebrafish embryos at 24 hpf and embryos injected with (<b>D–F</b>) <i>miR-3906</i> dsR (1.5 ng), (<b>G–I</b>) <i>miR-3906</i>-MO (8 ng), (<b>J–L</b>) <i>homer-1b</i> mRNA (400 pg), and (<b>M–O</b>) <i>homer-1b</i>-MO (3 ng) were collected, and the expressions of fast muscle type <i>fmhc4</i>, slow muscle type <i>smhc1</i>, and calcium-sensitive gene <i>atp2a1</i> were detected by WISH. (<b>P–R</b>) Embryos soaked with caffeine served as a positive control, indicating that [Ca<sup>2+</sup>]<sub>i</sub> was increased. (<b>S–U</b>) Embryos soaked with 2-APB served as a negative control, indicating that [Ca<sup>2+</sup>]<sub>i</sub> was reduced. Compared to WT (<b>A</b>), the expression of <i>fmhc4</i> in the embryos injected with <i>miR-3906</i>-MO (<b>G</b>), <i>homer-1b</i>-mRNA (<b>J</b>) and soaked in caffeine (<b>P</b>) was increased. The expression of <i>fmhc4</i> in the embryos injected with <i>miR-3906</i> dsR (<b>D</b>), <i>homer-1b</i>-MO (<b>M</b>) and soaked in 2-APB (<b>S</b>) was decreased. The expressions of <i>smhc1</i> in the embryos injected with <i>miR-3906</i> and <i>homer-1b</i> showed no difference (<b>B, E, H</b> and <b>K</b>), while <i>smhc1</i> expression was increased in embryos soaked in caffeine (<b>Q</b>) and reduced in embryos soaked in 2-APB (<b>T</b>). The expression patterns of <i>atp2a1</i> (<b>F, I, L, O, R</b> and <b>U</b>) in the examined embryos displayed a result similar to that of <i>fmhc4.</i> (<b>V</b>) The expression levels of the examined genes were quantified by q-PCR analysis after embryos were treated as indicated. Three rescue experiments were included: co-injection of pHsp-<i>wob-homer-1b</i> (lanes 9 and 10), soaking with caffeine (lanes 12 and 13) and incubating with 2-APB (lanes 15 and 16). SigmaPlot software was used to perform the Student’s <i>t</i>-test. *: indicates the difference at <i>P<0.05</i> level; **: indicates the difference at <i>P<0.01</i> level. The numbers shown in the lower-right corner of panels A–U indicate the number of phenotypes out of the total number of embryos examined.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "mrna", "differentiation", "regulating", "calcium"], "article_id"=>760753, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0070187.g006", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Injection_of_either_miR_3906_MO_or_homer_1b_mRNA_affects_fast_muscle_differentiation_by_regulating_the_calcium_concentration_in_muscle_cells_/760753", "title"=>"Injection of either <i>miR-3906-</i>MO or <i>homer-1b</i> mRNA affects fast muscle differentiation by regulating the calcium concentration in muscle cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-31 01:50:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1138345"], "description"=>"<p>(<b>A</b>) One-cell embryos were injected with buffer (A, as a control group WT), (<b>B</b>) <i>miR-3906</i> dsR (1.15 ng), (<b>C</b>) <i>miR-3906</i>-MO (8 ng), (<b>D</b>) <i>homer-1b</i> mRNA (400 pg) or (<b>E</b>) <i>homer-1b</i>-MO (3 ng) with calcium green-1 and tetramethylrhodamine (as an internal control). The gradient distribution of Ca<sup>2+</sup> concentration within the muscle cells in somites was analyzed at 24 hpf by ImageJ software analysis. There were 16 colors of signal images displaying the different intracellular Ca<sup>2+</sup> concentrations from low to high. We sketched a fixed region comprised of 11 to 20 trunk somites, as indicated, to calculate the readings from excitation emission (the region was indicated by white line). (<b>F</b>) After normalization, the reading from the WT group was set as 100, and the readings of <i>miR-3906</i> dsR-injected, <i>miR-3906</i>-MO-injected, <i>homer-1b-</i>mRNA-injected and <i>homer-1b-</i>MO-injected groups were compared. (<b>G</b>) The primary culture cells were stained with Fura2-AM, excited with 380 nm light, with emission at 510 nm. (<b>H</b>) The primary culture cells which displayed red fluorescence were the fast muscle cells. (<b>I</b>) The merge picture of F and G. Cells which had overlapping signals denoted in yellow were selected as detection samples (indicated by red box). (<b>J</b>) The image of fluorescence intensity shown on fast muscle cells that were excited by 340 nm, with emission at 510 nm. (<b>K</b>) The image of fluorescence intensity shown on fast muscle cells that were excited by 380 nm, with emission at 510 nm. (<b>L</b>) The value of [Ca<sup>2+</sup>]<sub>i</sub> was calculated by following the formula established by Grynkiewicz <i>et al</i>. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070187#pone.0070187-Myhre1\" target=\"_blank\">[31]</a> (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070187#s2\" target=\"_blank\">Materials and Methods</a>). The values of [Ca<sup>2+</sup>]<sub>i</sub> were obtained from cells derived from WT, <i>miR-3906</i> dsR-injected, <i>miR-3906</i>-MO-injected, <i>homer-1b</i>-mRNA-injected, <i>homer-1b</i>-MO-injected, Photo-<i>homer-1b</i>-MO-injected, caffeine-soaked and 2-APB-soaked embryos, followed by heat-shocked WT, pHsp-<i>miR-3906</i>-injected and pHsp-<i>wob-homer-1b</i>-injected embryos. SigmaPlot software was used to perform Student’s <i>t</i>-test. *: indicates the difference at <i>P<0.05</i> level; **: indicates the difference at <i>P<0.01</i> level. Ex: excitation; Em: emission.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "calcium"], "article_id"=>760752, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0070187.g005", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Injection_of_either_homer1b_mRNA_or_miR_3906_MO_increases_the_calcium_concentration_within_muscle_cells_/760752", "title"=>"Injection of either <i>homer1b-</i>mRNA or <i>miR-3906-</i>MO increases the calcium concentration within muscle cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-31 01:50:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1138358"], "description"=>"<p>Overexpression or knockdown of either <i>miR-3096</i> or <i>homer-1b</i> causes an imbalance between <i>miR-3096</i> and <i>homer-1b,</i> resulting in disturbing [Ca<sup>2+</sup>]<sub>i</sub> homeostasis in the fast muscle cells during differentiation. At this developmental stage, <i>miR-3906</i>, which is transcribed from its own promoter, decreases [Ca<sup>2+</sup>]<sub>i</sub> through inhibiting the translation of <i>homer-1b</i> mRNA and causing the reduction of Homer-1b protein. This fine-tuning of the protein amount of Homer-1b by <i>miR-3096</i> allows <i>homer-1b</i> to be present at a normal level in order to maintain [Ca<sup>2+</sup>]<sub>i</sub> homeostasis, thus ensuring normal fast muscle differentiation.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "illustrating", "controls", "homeostasis", "cells"], "article_id"=>760765, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0070187.g009", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_model_illustrating_how_miR_3906_controls_Ca_2_i_homeostasis_in_fast_muscle_cells_during_differentiation_/760765", "title"=>"Schematic model illustrating how <i>miR-3906</i> controls [Ca<sup>2+</sup>]<sub>i</sub> homeostasis in fast muscle cells during differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-31 01:50:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1138340"], "description"=>"<p>(<b>A</b>–<b>I</b>) Embryos derived from transgenic line Tg (α-actin:RFP), in which red fluorescence protein (RFP) is expressed specifically in skeletal muscles, were injected with <i>miR-3906</i>-MO, <i>miR-3906</i> dsR (mature double- strand <i>miR-3906</i>), <i>homer-1b</i>-MO or <i>homer-1b</i> mRNA, as indicated. After injection, the development of trunk muscle was observed at 32 hpf. Compared with WT (<b>A</b>), the morphological trait of embryos injected with either <i>miR-3906</i>-MO (<b>B</b>) or <i>homer-1b</i> mRNA (<b>C</b>) was similar to that of WT. Embryos injected with <i>homer-1b</i>-MO (<b>D</b>–<b>F</b>) and exogenous <i>miR-3906</i> (<i>miR-3906</i> dsR) (<b>G</b>–<b>I</b>) displayed such defective phenotypes as body axis bending and trunk muscle shortening in tail. Based on the definition of three defective levels of muscle (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070187#s2\" target=\"_blank\">Materials and Methods</a>), we calculated the defective percentages of embryos injected with <i>homer-1b</i>-MO alone (<b>J</b>) and <i>homer-1b</i>-MO plus wobble <i>homer-1b</i> mRNA (<b>K</b>). The occurrence percentages of severe and mild defects of embryos injected with 1.15 ng <i>miR-3906</i> dsR alone, 1.15 ng <i>miR-3906</i> dsR plus 3 ng <i>homer-1b</i>-MO and 1.15 ng <i>miR-3906</i> dsR plus 400 pg <i>homer-1b</i> mRNA were calculated (<b>L</b>). n: indicates the total number of embryos after three injections. (<b>M</b>) Western blot analysis of Homer-1b (48 kD, arrow) in WT (lanes 1 and 4), 3 ng <i>homer-1b</i>-MO-injected embryos (lane 2), 8 ng <i>miR-3906</i>-MO-injected embryos (lane 3) and 1.15 ng <i>miR-3906</i> dsR-injected embryos (lane 5). Detection of α-tubulin served as internal control (arrowhead). The relative intensity of Homer-1b protein among WT, <i>homer-1b</i>-MO, <i>miR-3906</i>-MO and <i>miR-3906</i> dsR was also indicated.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "phenotypes", "induced", "injection"], "article_id"=>760747, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0070187.g004", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Defective_phenotypes_induced_by_injection_of_excessive_miR_3906_and_homer_1b_MO_were_similar_/760747", "title"=>"Defective phenotypes induced by injection of excessive <i>miR-3906</i> and <i>homer-1b</i>-MO were similar.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-31 01:50:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1138354"], "description"=>"<p>Using phalloidin stain, compared to (<b>A</b>) WT embryos, (<b>B</b>) <i>miR-3906-</i>MO-injected embryos, (<b>C</b>) <i>homer-1b-</i>mRNA-injected embryos and (<b>D</b>) caffeine-soaked embryos exhibited a bending and disruption of sarcomeric actin organization in muscle cells. In longitudinal section, compared to (<b>E</b>) WT control embryos, (<b>F</b>) the Z-disc structure displaying in <i>miR-3906-</i>MO-injected embryos, (<b>G</b>) <i>homer-1b-</i>mRNA-injected embryos and (<b>H</b>) caffeine-soaked embryos was chaotic (indicated by arrows), and the full-length sarcomeres in muscle fibers were interrupted (indicated by asterisks) and could not be seen completely in a full sarcomere line. In cross section, compared to (<b>I</b>) WT control embryos, the relative position and proportion between myosin heavy chain and actin filament among the (<b>J</b>) <i>miR-3906-</i>MO-injected embryos, (<b>K</b>) <i>homer-1b-</i>mRNA-injected embryos and (<b>L</b>) caffeine-soaked embryos were chaotic and extremely irregular. For each sample, we randomly selected five positions to identify their hexagonal arrangements and calculated the ratio between the thick and thin filaments (n = 3). Results were presented at the bottom right of each picture (<b>I–L</b>). M:M line;Z:Z-disc;scale bar:0.1 µm.</p>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "inhibition", "overexpression", "disrupts", "sarcomeric", "actin"], "article_id"=>760761, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0070187.g007", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Either_inhibition_of_miR_3906_or_overexpression_of_homer_1b_disrupts_sarcomeric_actin_organization_/760761", "title"=>"Either inhibition of <i>miR-3906</i> or overexpression of <i>homer-1b</i> disrupts sarcomeric actin organization.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-31 01:50:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1138366", "https://ndownloader.figshare.com/files/1138377"], "description"=>"<div><p>A microRNA, termed <i>miR-In300</i> or <i>miR-3906</i>, suppresses the transcription of <i>myf5</i> through silencing <i>dickkopf-related protein 3</i> (<i>dkk3r/dkk3a</i>) during early development when <i>myf5</i> is highly transcribed, but not at late stages when <i>myf5</i> transcription is reduced. Moreover, after 24 hpf, when muscle cells are starting to differentiate, Dkk3a could not be detected in muscle tissue at 20 hpf. To explain these reversals, we collected embryos at 32 hpf, performed assays, and identified <i>homer-1b</i>, which regulates calcium release from sarcoplasmic reticulum, as the target gene of <i>miR-3906</i>. We further found that either <i>miR-3906</i> knockdown or <i>homer-1b</i> overexpression increased expressions of <i>fmhc4</i> and <i>atp2a1</i> of calcium-dependent fast muscle fibrils, but not slow muscle fibrils, and caused a severe disruption of sarcomeric actin and Z-disc structure. Additionally, compared to control embryos, the intracellular calcium concentration ([Ca<sup>2+</sup>]<sub>i</sub>) of these treated embryos was increased as high as 83.9–97.3% in fast muscle. In contrast, either <i>miR-3906</i> overexpression or <i>homer-1b</i> knockdown caused decreases of [Ca<sup>2+</sup>]<sub>i</sub> and, correspondingly, defective phenotypes in fast muscle. These defects could be rescued by inducing <i>homer-1b</i> expression at later stage. These results indicate that <i>miR-3906</i> controls [Ca<sup>2+</sup>]<sub>i</sub> homeostasis in fast muscle through fine tuning <i>homer-1b</i> expression during differentiation to maintain normal muscle development.</p></div>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "regulates", "differentiation", "modulating", "zebrafish", "embryos"], "article_id"=>760773, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0070187.s001", "https://dx.doi.org/10.1371/journal.pone.0070187.s002"], "stats"=>{"downloads"=>4, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MicroRNA_3906_Regulates_Fast_Muscle_Differentiation_through_Modulating_the_Target_Gene_homer_1b_in_Zebrafish_Embryos/760773", "title"=>"<i>MicroRNA-3906</i> Regulates Fast Muscle Differentiation through Modulating the Target Gene <i>homer-1b</i> in Zebrafish Embryos", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-07-31 01:50:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1138334"], "description"=>"<p>(<b>A</b>) Constructs for examining <i>luc</i> assay. The complete 3′UTR segments of <i>col1a2</i>, <i>dnajc10</i>, <i>homer-1b</i>, <i>six1a</i> and <i>trmt2a,</i> which are putative target genes of <i>miR-3906,</i> were ligated into the downstream of <i>luc</i> reporter gene contained in plasmid phRG-TK. (<b>B</b>) Plasmid pCMV-RFP-<i>miR-3906</i> [<i>miR-3906</i> (+)], a RFP reporter fused with pre-<i>miR-3906</i> and driven by CMV promoter, was co-transfected with pGL3-TK (internal control) and each examined construct, as indicated, into HEK 293T cells. Injection of pGL3-TK and each examined construct without containing pCMV-RFP-<i>miR-3906</i> [<i>miR-3906</i> (−)] served as a control group, and its <i>luc</i> activity was 100%. In zebrafish embryos, we co-injected synthetic pre-<i>miR-3906</i> RNA [miR-3906 (+)], pGL3-TK (internal control) and each examined construct in the experimental group. Injection of pGL3-TK and each examined construct without containing pre-<i>miR-3906</i> RNA [miR-3906 (−)] served as control group, and its <i>luc</i> activity was 100%. Data were presented as means±SD from three independent experiments (n = 3). (<b>C</b>) Injection of plasmid phRG-TK, in which <i>luc</i> expression was driven by thymidine kinase (TK), served as a control, and its <i>luc</i> activity was 100%. The relative <i>luc</i> expression level of each combination, as indicated, was examined. All data were presented as means±SD from three independent experiments (n = 3). (<b>D</b>) Various lengths of 3′UTR segment derived from <i>homer-1b</i> mRNA (from 1198 to 2222 nt) fused with <i>luc</i> reporter gene and driven by TK promoter were constructed as indicated. Plasmid alone or plasmid plus pre-<i>miR-3906</i> were individually injected in zebrafish embryos to perform <i>luc</i> assay. Data were presented as means±SD from three independent experiments (n = 3). Student's <i>t</i>-test determined significant differences between each group, and * indicates that the difference was significant at <i>P</i><0.05. (black box: <i>miR-3906-</i>target sequences; cross filled box: <i>miR-3906</i>-target mutated sequences).</p>", "links"=>[], "tags"=>["Anatomy and physiology", "Musculoskeletal system", "muscle", "developmental biology", "Cell differentiation", "genetics", "Model organisms", "Molecular cell biology", "assay", "co-injected", "plasmid", "constructs", "containing", "putative"], "article_id"=>760741, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Cheng-Yung Lin", "Jie-Shin Chen", "Moo-Rung Loo", "Chung-Ching Hsiao", "Wen-Yen Chang", "Huai-Jen Tsai"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0070187.g001", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Luciferase_luc_activity_assay_of_miR_3906_co_injected_with_plasmid_constructs_containing_3_UTR_segment_of_putative_miR_3906_target_genes_/760741", "title"=>"Luciferase (<i>luc</i>) activity assay of <i>miR-3906</i> co-injected with plasmid constructs containing 3′UTR segment of putative <i>miR-3906</i>-target genes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-31 01:50:49"}

PMC Usage Stats | Further Information

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Relative Metric

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