Fluorescent-Based Methods for Gene Knockdown and Functional Cardiac Imaging in Zebrafish
A notable advantage of zebrafish as a model organism is the ease of gene knockdown using morpholino antisense oligonucleotide (MO). However, zebrafish morphants injected with MO for a target protein often show heterogeneous phenotypes, despite controlling the injection volume of the MO solution in a...
Gespeichert in:
| Veröffentlicht in: | Molecular biotechnology 2013-10, Vol.55 (2), p.131-142 |
|---|---|
| Hauptverfasser: | , , , , , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 142 |
|---|---|
| container_issue | 2 |
| container_start_page | 131 |
| container_title | Molecular biotechnology |
| container_volume | 55 |
| creator | Umemoto, Noriko Nishimura, Yuhei Shimada, Yasuhito Yamanaka, Yukiko Kishi, Seiya Ito, Saki Okamori, Kana Nakamura, Yuuki Kuroyanagi, Junya Zhang, Zi Zang, Liqing Wang, Zhipeng Nishimura, Norihiro Tanaka, Toshio |
| description | A notable advantage of zebrafish as a model organism is the ease of gene knockdown using morpholino antisense oligonucleotide (MO). However, zebrafish morphants injected with MO for a target protein often show heterogeneous phenotypes, despite controlling the injection volume of the MO solution in all embryos. We developed a method for estimating the quantity of MO injected into each living morphant, based on the co-injection of a control MO labeled with the fluorophore lissamine. By applying this method for knockdown of cardiac troponin T (tnnt2a) in zebrafish, we could efficiently select the partial tnnt2a-depleted zebrafish with a decreased heart rate and impairment of cardiac contraction. To investigate cardiac impairment of the tnnt2a morphant, we performed fluorescent cardiac imaging using Bodipy-ceramide. Cardiac image analysis showed moderate reduction of tnnt2a impaired diastolic distensibility and decreased contraction and relaxation velocities. To the best of our knowledge, this is the first report to analyze the role of tnnt2a in cardiac function in tnnt2a-depleted living animals. Our combinatorial approach can be applied for analyzing the molecular function of any protein associated with human cardiac diseases. |
| doi_str_mv | 10.1007/s12033-013-9664-6 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3825550</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1449769453</sourcerecordid><originalsourceid>FETCH-LOGICAL-c593t-e80ea48c8e5cc7bc57b114fdf098d381bed0735735c66227b2e787f4bf3a73cc3</originalsourceid><addsrcrecordid>eNqFkU9rFTEUxYMotlY_gBsNuOlm9Ob_zEaoj762tOJCC-ImZDKZeanzkprMKH5785i21C4UAgnc3z255x6EXhJ4SwDUu0woMFYBYVUjJa_kI7RPhGgqYCAelzcoVkmoxR56lvMVACWCs6dojzKpOMhmH31dj3NMLlsXpuqDya7DH920iV3GfUz4xAWHz0O037v4K2ATOryeg518DGbEK5M6byw-25rBhwH7gL-5Npne581z9KQ3Y3Yvbu4DdLk-_rI6rS4-nZytji4qKxo2Va4GZ3htayesVa0VqiWE910PTd2xmrSuKyZEOVZKSlVLnapVz9ueGcWsZQfo_aJ7Pbdb1-18JDPq6-S3Jv3W0Xj9dyX4jR7iT81qKoSAInB4I5Dij9nlSW99Wcc4muDinDWRkkkOhJL_o5w3SjZcsIK-eYBexTmVpS0UFZJCXSiyUDbFnJPr7-YmoHcR6yViXSLWu4i1LD2v7hu-67jNtAB0AXIphcGle1__Q_X10tSbqM2QfNaXnykQAVCSKCOzP3UHuls</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1449256208</pqid></control><display><type>article</type><title>Fluorescent-Based Methods for Gene Knockdown and Functional Cardiac Imaging in Zebrafish</title><source>MEDLINE</source><source>Springer Nature - Complete Springer Journals</source><creator>Umemoto, Noriko ; Nishimura, Yuhei ; Shimada, Yasuhito ; Yamanaka, Yukiko ; Kishi, Seiya ; Ito, Saki ; Okamori, Kana ; Nakamura, Yuuki ; Kuroyanagi, Junya ; Zhang, Zi ; Zang, Liqing ; Wang, Zhipeng ; Nishimura, Norihiro ; Tanaka, Toshio</creator><creatorcontrib>Umemoto, Noriko ; Nishimura, Yuhei ; Shimada, Yasuhito ; Yamanaka, Yukiko ; Kishi, Seiya ; Ito, Saki ; Okamori, Kana ; Nakamura, Yuuki ; Kuroyanagi, Junya ; Zhang, Zi ; Zang, Liqing ; Wang, Zhipeng ; Nishimura, Norihiro ; Tanaka, Toshio</creatorcontrib><description>A notable advantage of zebrafish as a model organism is the ease of gene knockdown using morpholino antisense oligonucleotide (MO). However, zebrafish morphants injected with MO for a target protein often show heterogeneous phenotypes, despite controlling the injection volume of the MO solution in all embryos. We developed a method for estimating the quantity of MO injected into each living morphant, based on the co-injection of a control MO labeled with the fluorophore lissamine. By applying this method for knockdown of cardiac troponin T (tnnt2a) in zebrafish, we could efficiently select the partial tnnt2a-depleted zebrafish with a decreased heart rate and impairment of cardiac contraction. To investigate cardiac impairment of the tnnt2a morphant, we performed fluorescent cardiac imaging using Bodipy-ceramide. Cardiac image analysis showed moderate reduction of tnnt2a impaired diastolic distensibility and decreased contraction and relaxation velocities. To the best of our knowledge, this is the first report to analyze the role of tnnt2a in cardiac function in tnnt2a-depleted living animals. Our combinatorial approach can be applied for analyzing the molecular function of any protein associated with human cardiac diseases.</description><identifier>ISSN: 1073-6085</identifier><identifier>EISSN: 1559-0305</identifier><identifier>DOI: 10.1007/s12033-013-9664-6</identifier><identifier>PMID: 23674069</identifier><language>eng</language><publisher>Boston: Springer-Verlag</publisher><subject>administration & dosage ; Animals ; Biochemistry ; Biological Techniques ; Biotechnology ; Cardiac Imaging Techniques ; cardiac output ; Cardiomyopathy ; Cell Biology ; Chemistry ; Chemistry and Materials Science ; Danio rerio ; Embryo, Nonmammalian ; embryology ; Fluorescence ; Fluorescent Dyes ; Gene Knockdown Techniques ; genes ; Genetics ; Heart ; Heart - physiology ; Heart Rate ; Human Genetics ; Humans ; image analysis ; Injection ; metabolism ; Models, Animal ; Morpholinos ; Morpholinos - administration & dosage ; Morpholinos - metabolism ; Mutation ; Myocardial Contraction ; Oligonucleotides, Antisense ; Oligonucleotides, Antisense - administration & dosage ; Oligonucleotides, Antisense - metabolism ; Phenotype ; physiology ; Protein Science ; troponin T ; Troponin T - genetics ; Troponin T - metabolism ; Zebrafish ; Zebrafish - embryology ; Zebrafish - genetics ; Zebrafish - metabolism ; Zebrafish Proteins ; Zebrafish Proteins - genetics ; Zebrafish Proteins - metabolism</subject><ispartof>Molecular biotechnology, 2013-10, Vol.55 (2), p.131-142</ispartof><rights>The Author(s) 2013</rights><rights>Springer Science+Business Media New York 2013</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c593t-e80ea48c8e5cc7bc57b114fdf098d381bed0735735c66227b2e787f4bf3a73cc3</citedby><cites>FETCH-LOGICAL-c593t-e80ea48c8e5cc7bc57b114fdf098d381bed0735735c66227b2e787f4bf3a73cc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s12033-013-9664-6$$EPDF$$P50$$Gspringer$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s12033-013-9664-6$$EHTML$$P50$$Gspringer$$Hfree_for_read</linktohtml><link.rule.ids>230,314,776,780,881,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23674069$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Umemoto, Noriko</creatorcontrib><creatorcontrib>Nishimura, Yuhei</creatorcontrib><creatorcontrib>Shimada, Yasuhito</creatorcontrib><creatorcontrib>Yamanaka, Yukiko</creatorcontrib><creatorcontrib>Kishi, Seiya</creatorcontrib><creatorcontrib>Ito, Saki</creatorcontrib><creatorcontrib>Okamori, Kana</creatorcontrib><creatorcontrib>Nakamura, Yuuki</creatorcontrib><creatorcontrib>Kuroyanagi, Junya</creatorcontrib><creatorcontrib>Zhang, Zi</creatorcontrib><creatorcontrib>Zang, Liqing</creatorcontrib><creatorcontrib>Wang, Zhipeng</creatorcontrib><creatorcontrib>Nishimura, Norihiro</creatorcontrib><creatorcontrib>Tanaka, Toshio</creatorcontrib><title>Fluorescent-Based Methods for Gene Knockdown and Functional Cardiac Imaging in Zebrafish</title><title>Molecular biotechnology</title><addtitle>Mol Biotechnol</addtitle><addtitle>Mol Biotechnol</addtitle><description>A notable advantage of zebrafish as a model organism is the ease of gene knockdown using morpholino antisense oligonucleotide (MO). However, zebrafish morphants injected with MO for a target protein often show heterogeneous phenotypes, despite controlling the injection volume of the MO solution in all embryos. We developed a method for estimating the quantity of MO injected into each living morphant, based on the co-injection of a control MO labeled with the fluorophore lissamine. By applying this method for knockdown of cardiac troponin T (tnnt2a) in zebrafish, we could efficiently select the partial tnnt2a-depleted zebrafish with a decreased heart rate and impairment of cardiac contraction. To investigate cardiac impairment of the tnnt2a morphant, we performed fluorescent cardiac imaging using Bodipy-ceramide. Cardiac image analysis showed moderate reduction of tnnt2a impaired diastolic distensibility and decreased contraction and relaxation velocities. To the best of our knowledge, this is the first report to analyze the role of tnnt2a in cardiac function in tnnt2a-depleted living animals. Our combinatorial approach can be applied for analyzing the molecular function of any protein associated with human cardiac diseases.</description><subject>administration & dosage</subject><subject>Animals</subject><subject>Biochemistry</subject><subject>Biological Techniques</subject><subject>Biotechnology</subject><subject>Cardiac Imaging Techniques</subject><subject>cardiac output</subject><subject>Cardiomyopathy</subject><subject>Cell Biology</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Danio rerio</subject><subject>Embryo, Nonmammalian</subject><subject>embryology</subject><subject>Fluorescence</subject><subject>Fluorescent Dyes</subject><subject>Gene Knockdown Techniques</subject><subject>genes</subject><subject>Genetics</subject><subject>Heart</subject><subject>Heart - physiology</subject><subject>Heart Rate</subject><subject>Human Genetics</subject><subject>Humans</subject><subject>image analysis</subject><subject>Injection</subject><subject>metabolism</subject><subject>Models, Animal</subject><subject>Morpholinos</subject><subject>Morpholinos - administration & dosage</subject><subject>Morpholinos - metabolism</subject><subject>Mutation</subject><subject>Myocardial Contraction</subject><subject>Oligonucleotides, Antisense</subject><subject>Oligonucleotides, Antisense - administration & dosage</subject><subject>Oligonucleotides, Antisense - metabolism</subject><subject>Phenotype</subject><subject>physiology</subject><subject>Protein Science</subject><subject>troponin T</subject><subject>Troponin T - genetics</subject><subject>Troponin T - metabolism</subject><subject>Zebrafish</subject><subject>Zebrafish - embryology</subject><subject>Zebrafish - genetics</subject><subject>Zebrafish - metabolism</subject><subject>Zebrafish Proteins</subject><subject>Zebrafish Proteins - genetics</subject><subject>Zebrafish Proteins - metabolism</subject><issn>1073-6085</issn><issn>1559-0305</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqFkU9rFTEUxYMotlY_gBsNuOlm9Ob_zEaoj762tOJCC-ImZDKZeanzkprMKH5785i21C4UAgnc3z255x6EXhJ4SwDUu0woMFYBYVUjJa_kI7RPhGgqYCAelzcoVkmoxR56lvMVACWCs6dojzKpOMhmH31dj3NMLlsXpuqDya7DH920iV3GfUz4xAWHz0O037v4K2ATOryeg518DGbEK5M6byw-25rBhwH7gL-5Npne581z9KQ3Y3Yvbu4DdLk-_rI6rS4-nZytji4qKxo2Va4GZ3htayesVa0VqiWE910PTd2xmrSuKyZEOVZKSlVLnapVz9ueGcWsZQfo_aJ7Pbdb1-18JDPq6-S3Jv3W0Xj9dyX4jR7iT81qKoSAInB4I5Dij9nlSW99Wcc4muDinDWRkkkOhJL_o5w3SjZcsIK-eYBexTmVpS0UFZJCXSiyUDbFnJPr7-YmoHcR6yViXSLWu4i1LD2v7hu-67jNtAB0AXIphcGle1__Q_X10tSbqM2QfNaXnykQAVCSKCOzP3UHuls</recordid><startdate>20131001</startdate><enddate>20131001</enddate><creator>Umemoto, Noriko</creator><creator>Nishimura, Yuhei</creator><creator>Shimada, Yasuhito</creator><creator>Yamanaka, Yukiko</creator><creator>Kishi, Seiya</creator><creator>Ito, Saki</creator><creator>Okamori, Kana</creator><creator>Nakamura, Yuuki</creator><creator>Kuroyanagi, Junya</creator><creator>Zhang, Zi</creator><creator>Zang, Liqing</creator><creator>Wang, Zhipeng</creator><creator>Nishimura, Norihiro</creator><creator>Tanaka, Toshio</creator><general>Springer-Verlag</general><general>Springer US</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>C6C</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>Q9U</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20131001</creationdate><title>Fluorescent-Based Methods for Gene Knockdown and Functional Cardiac Imaging in Zebrafish</title><author>Umemoto, Noriko ; Nishimura, Yuhei ; Shimada, Yasuhito ; Yamanaka, Yukiko ; Kishi, Seiya ; Ito, Saki ; Okamori, Kana ; Nakamura, Yuuki ; Kuroyanagi, Junya ; Zhang, Zi ; Zang, Liqing ; Wang, Zhipeng ; Nishimura, Norihiro ; Tanaka, Toshio</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c593t-e80ea48c8e5cc7bc57b114fdf098d381bed0735735c66227b2e787f4bf3a73cc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>administration & dosage</topic><topic>Animals</topic><topic>Biochemistry</topic><topic>Biological Techniques</topic><topic>Biotechnology</topic><topic>Cardiac Imaging Techniques</topic><topic>cardiac output</topic><topic>Cardiomyopathy</topic><topic>Cell Biology</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Danio rerio</topic><topic>Embryo, Nonmammalian</topic><topic>embryology</topic><topic>Fluorescence</topic><topic>Fluorescent Dyes</topic><topic>Gene Knockdown Techniques</topic><topic>genes</topic><topic>Genetics</topic><topic>Heart</topic><topic>Heart - physiology</topic><topic>Heart Rate</topic><topic>Human Genetics</topic><topic>Humans</topic><topic>image analysis</topic><topic>Injection</topic><topic>metabolism</topic><topic>Models, Animal</topic><topic>Morpholinos</topic><topic>Morpholinos - administration & dosage</topic><topic>Morpholinos - metabolism</topic><topic>Mutation</topic><topic>Myocardial Contraction</topic><topic>Oligonucleotides, Antisense</topic><topic>Oligonucleotides, Antisense - administration & dosage</topic><topic>Oligonucleotides, Antisense - metabolism</topic><topic>Phenotype</topic><topic>physiology</topic><topic>Protein Science</topic><topic>troponin T</topic><topic>Troponin T - genetics</topic><topic>Troponin T - metabolism</topic><topic>Zebrafish</topic><topic>Zebrafish - embryology</topic><topic>Zebrafish - genetics</topic><topic>Zebrafish - metabolism</topic><topic>Zebrafish Proteins</topic><topic>Zebrafish Proteins - genetics</topic><topic>Zebrafish Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Umemoto, Noriko</creatorcontrib><creatorcontrib>Nishimura, Yuhei</creatorcontrib><creatorcontrib>Shimada, Yasuhito</creatorcontrib><creatorcontrib>Yamanaka, Yukiko</creatorcontrib><creatorcontrib>Kishi, Seiya</creatorcontrib><creatorcontrib>Ito, Saki</creatorcontrib><creatorcontrib>Okamori, Kana</creatorcontrib><creatorcontrib>Nakamura, Yuuki</creatorcontrib><creatorcontrib>Kuroyanagi, Junya</creatorcontrib><creatorcontrib>Zhang, Zi</creatorcontrib><creatorcontrib>Zang, Liqing</creatorcontrib><creatorcontrib>Wang, Zhipeng</creatorcontrib><creatorcontrib>Nishimura, Norihiro</creatorcontrib><creatorcontrib>Tanaka, Toshio</creatorcontrib><collection>AGRIS</collection><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Engineering Collection</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Molecular biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Umemoto, Noriko</au><au>Nishimura, Yuhei</au><au>Shimada, Yasuhito</au><au>Yamanaka, Yukiko</au><au>Kishi, Seiya</au><au>Ito, Saki</au><au>Okamori, Kana</au><au>Nakamura, Yuuki</au><au>Kuroyanagi, Junya</au><au>Zhang, Zi</au><au>Zang, Liqing</au><au>Wang, Zhipeng</au><au>Nishimura, Norihiro</au><au>Tanaka, Toshio</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Fluorescent-Based Methods for Gene Knockdown and Functional Cardiac Imaging in Zebrafish</atitle><jtitle>Molecular biotechnology</jtitle><stitle>Mol Biotechnol</stitle><addtitle>Mol Biotechnol</addtitle><date>2013-10-01</date><risdate>2013</risdate><volume>55</volume><issue>2</issue><spage>131</spage><epage>142</epage><pages>131-142</pages><issn>1073-6085</issn><eissn>1559-0305</eissn><abstract>A notable advantage of zebrafish as a model organism is the ease of gene knockdown using morpholino antisense oligonucleotide (MO). However, zebrafish morphants injected with MO for a target protein often show heterogeneous phenotypes, despite controlling the injection volume of the MO solution in all embryos. We developed a method for estimating the quantity of MO injected into each living morphant, based on the co-injection of a control MO labeled with the fluorophore lissamine. By applying this method for knockdown of cardiac troponin T (tnnt2a) in zebrafish, we could efficiently select the partial tnnt2a-depleted zebrafish with a decreased heart rate and impairment of cardiac contraction. To investigate cardiac impairment of the tnnt2a morphant, we performed fluorescent cardiac imaging using Bodipy-ceramide. Cardiac image analysis showed moderate reduction of tnnt2a impaired diastolic distensibility and decreased contraction and relaxation velocities. To the best of our knowledge, this is the first report to analyze the role of tnnt2a in cardiac function in tnnt2a-depleted living animals. Our combinatorial approach can be applied for analyzing the molecular function of any protein associated with human cardiac diseases.</abstract><cop>Boston</cop><pub>Springer-Verlag</pub><pmid>23674069</pmid><doi>10.1007/s12033-013-9664-6</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1073-6085 |
| ispartof | Molecular biotechnology, 2013-10, Vol.55 (2), p.131-142 |
| issn | 1073-6085 1559-0305 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3825550 |
| source | MEDLINE; Springer Nature - Complete Springer Journals |
| subjects | administration & dosage Animals Biochemistry Biological Techniques Biotechnology Cardiac Imaging Techniques cardiac output Cardiomyopathy Cell Biology Chemistry Chemistry and Materials Science Danio rerio Embryo, Nonmammalian embryology Fluorescence Fluorescent Dyes Gene Knockdown Techniques genes Genetics Heart Heart - physiology Heart Rate Human Genetics Humans image analysis Injection metabolism Models, Animal Morpholinos Morpholinos - administration & dosage Morpholinos - metabolism Mutation Myocardial Contraction Oligonucleotides, Antisense Oligonucleotides, Antisense - administration & dosage Oligonucleotides, Antisense - metabolism Phenotype physiology Protein Science troponin T Troponin T - genetics Troponin T - metabolism Zebrafish Zebrafish - embryology Zebrafish - genetics Zebrafish - metabolism Zebrafish Proteins Zebrafish Proteins - genetics Zebrafish Proteins - metabolism |
| title | Fluorescent-Based Methods for Gene Knockdown and Functional Cardiac Imaging in Zebrafish |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-01T05%3A04%3A46IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Fluorescent-Based%20Methods%20for%20Gene%20Knockdown%20and%20Functional%20Cardiac%20Imaging%20in%20Zebrafish&rft.jtitle=Molecular%20biotechnology&rft.au=Umemoto,%20Noriko&rft.date=2013-10-01&rft.volume=55&rft.issue=2&rft.spage=131&rft.epage=142&rft.pages=131-142&rft.issn=1073-6085&rft.eissn=1559-0305&rft_id=info:doi/10.1007/s12033-013-9664-6&rft_dat=%3Cproquest_pubme%3E1449769453%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1449256208&rft_id=info:pmid/23674069&rfr_iscdi=true |