A new approach to manipulate the fate of single neural stem cells in tissue
This study describes a microinjection technique that allows for the acute manipulation of individual neural stem cells in organotypic slice cultures via direct delivery of biologically active molecules. A challenge in the field of neural stem cell biology is the mechanistic dissection of single stem...
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| Veröffentlicht in: | Nature neuroscience 2012-02, Vol.15 (2), p.329-337 |
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| creator | Taverna, Elena Haffner, Christiane Pepperkok, Rainer Huttner, Wieland B |
| description | This study describes a microinjection technique that allows for the acute manipulation of individual neural stem cells in organotypic slice cultures via direct delivery of biologically active molecules.
A challenge in the field of neural stem cell biology is the mechanistic dissection of single stem cell behavior in tissue. Although such behavior can be tracked by sophisticated imaging techniques, current methods of genetic manipulation do not allow researchers to change the level of a defined gene product on a truly acute time scale and are limited to very few genes at a time. To overcome these limitations, we established microinjection of neuroepithelial/radial glial cells (apical progenitors) in organotypic slice culture of embryonic mouse brain. Microinjected apical progenitors showed cell cycle parameters that were indistinguishable to apical progenitors
in utero
, underwent self-renewing divisions and generated neurons. Microinjection of single genes, recombinant proteins or complex mixtures of RNA was found to elicit acute and defined changes in apical progenitor behavior and progeny fate. Thus, apical progenitor microinjection provides a new approach to acutely manipulating single neural stem and progenitor cells in tissue. |
| doi_str_mv | 10.1038/nn.3008 |
| format | Article |
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A challenge in the field of neural stem cell biology is the mechanistic dissection of single stem cell behavior in tissue. Although such behavior can be tracked by sophisticated imaging techniques, current methods of genetic manipulation do not allow researchers to change the level of a defined gene product on a truly acute time scale and are limited to very few genes at a time. To overcome these limitations, we established microinjection of neuroepithelial/radial glial cells (apical progenitors) in organotypic slice culture of embryonic mouse brain. Microinjected apical progenitors showed cell cycle parameters that were indistinguishable to apical progenitors
in utero
, underwent self-renewing divisions and generated neurons. Microinjection of single genes, recombinant proteins or complex mixtures of RNA was found to elicit acute and defined changes in apical progenitor behavior and progeny fate. Thus, apical progenitor microinjection provides a new approach to acutely manipulating single neural stem and progenitor cells in tissue.</description><identifier>ISSN: 1097-6256</identifier><identifier>EISSN: 1546-1726</identifier><identifier>DOI: 10.1038/nn.3008</identifier><identifier>PMID: 22179113</identifier><identifier>CODEN: NANEFN</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>631/136 ; 631/378/2183/2182 ; 631/378/2571/2579 ; Animal Genetics and Genomics ; Animals ; Behavioral Sciences ; Biological Techniques ; Biomedical and Life Sciences ; Biomedicine ; cdc42 GTP-Binding Protein - administration & dosage ; cdc42 GTP-Binding Protein - metabolism ; Cell culture ; Cell Cycle - drug effects ; Cell Cycle - genetics ; Cell Differentiation - genetics ; Cell Differentiation - physiology ; Embryo, Mammalian ; Immediate-Early Proteins - genetics ; In Vitro Techniques ; Luminescent Proteins - administration & dosage ; Luminescent Proteins - genetics ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microinjections - methods ; Mutation - genetics ; Neural Stem Cells - drug effects ; Neural Stem Cells - physiology ; Neurobiology ; Neurosciences ; Physiological aspects ; Rhombencephalon - cytology ; Rhombencephalon - embryology ; RNA ; RNA, Messenger - pharmacology ; Stem cells ; technical-report ; Time Factors ; Tumor Suppressor Proteins - genetics</subject><ispartof>Nature neuroscience, 2012-02, Vol.15 (2), p.329-337</ispartof><rights>Springer Nature America, Inc. 2011</rights><rights>COPYRIGHT 2012 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Feb 2012</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c475t-8a976337cd45766ad6cee51103d8e55563106e27b7f592680abccea235a7386b3</citedby><cites>FETCH-LOGICAL-c475t-8a976337cd45766ad6cee51103d8e55563106e27b7f592680abccea235a7386b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nn.3008$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nn.3008$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22179113$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Taverna, Elena</creatorcontrib><creatorcontrib>Haffner, Christiane</creatorcontrib><creatorcontrib>Pepperkok, Rainer</creatorcontrib><creatorcontrib>Huttner, Wieland B</creatorcontrib><title>A new approach to manipulate the fate of single neural stem cells in tissue</title><title>Nature neuroscience</title><addtitle>Nat Neurosci</addtitle><addtitle>Nat Neurosci</addtitle><description>This study describes a microinjection technique that allows for the acute manipulation of individual neural stem cells in organotypic slice cultures via direct delivery of biologically active molecules.
A challenge in the field of neural stem cell biology is the mechanistic dissection of single stem cell behavior in tissue. Although such behavior can be tracked by sophisticated imaging techniques, current methods of genetic manipulation do not allow researchers to change the level of a defined gene product on a truly acute time scale and are limited to very few genes at a time. To overcome these limitations, we established microinjection of neuroepithelial/radial glial cells (apical progenitors) in organotypic slice culture of embryonic mouse brain. Microinjected apical progenitors showed cell cycle parameters that were indistinguishable to apical progenitors
in utero
, underwent self-renewing divisions and generated neurons. Microinjection of single genes, recombinant proteins or complex mixtures of RNA was found to elicit acute and defined changes in apical progenitor behavior and progeny fate. Thus, apical progenitor microinjection provides a new approach to acutely manipulating single neural stem and progenitor cells in tissue.</description><subject>631/136</subject><subject>631/378/2183/2182</subject><subject>631/378/2571/2579</subject><subject>Animal Genetics and Genomics</subject><subject>Animals</subject><subject>Behavioral Sciences</subject><subject>Biological Techniques</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>cdc42 GTP-Binding Protein - administration & dosage</subject><subject>cdc42 GTP-Binding Protein - metabolism</subject><subject>Cell culture</subject><subject>Cell Cycle - drug effects</subject><subject>Cell Cycle - genetics</subject><subject>Cell Differentiation - genetics</subject><subject>Cell Differentiation - physiology</subject><subject>Embryo, Mammalian</subject><subject>Immediate-Early Proteins - genetics</subject><subject>In Vitro Techniques</subject><subject>Luminescent Proteins - administration & dosage</subject><subject>Luminescent Proteins - genetics</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Mice, Transgenic</subject><subject>Microinjections - methods</subject><subject>Mutation - genetics</subject><subject>Neural Stem Cells - drug effects</subject><subject>Neural Stem Cells - physiology</subject><subject>Neurobiology</subject><subject>Neurosciences</subject><subject>Physiological aspects</subject><subject>Rhombencephalon - cytology</subject><subject>Rhombencephalon - embryology</subject><subject>RNA</subject><subject>RNA, Messenger - pharmacology</subject><subject>Stem cells</subject><subject>technical-report</subject><subject>Time Factors</subject><subject>Tumor Suppressor Proteins - genetics</subject><issn>1097-6256</issn><issn>1546-1726</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kttq3DAQhkVoyKklb1AEvWh64a0O1sGXS0jSkEChh2uhlccbBVveSjJN374y3SbZUIouNEjf_Jp_NAidUrKghOuPISw4IXoPHVFRy4oqJl-VmDSqkkzIQ3Sc0j0hRAndHKBDxqhqKOVH6GaJA_zEdrOJo3V3OI94sMFvpt5mwPkOcDcHY4eTD-seCj1F2-OUYcAO-j5hH3D2KU3wGu13tk_wZrufoO-XF9_OP1W3n6-uz5e3lauVyJW2jZKcK9fWQklpW-kABC0-Wg1CCMkpkcDUSnWiYVITu3IOLOPCKq7lip-g9390S80_JkjZDD7NtdgA45RMQ7VQQlFeyLP_kpTQmhJR07qg716g9-MUQ_ExU0Iy0uhn1Nr2YHzoxhytm0XNkunS4FqL-dnFP6iyWhi8GwN0vpzvJHzYSShMhoe8tlNK5vrrl112697FMaUIndlEP9j4q9Rp5mEwIZh5GAr5dmtpWg3QPnJ_f_-pPalchTXE5553tX4Dt1K2yg</recordid><startdate>20120201</startdate><enddate>20120201</enddate><creator>Taverna, Elena</creator><creator>Haffner, Christiane</creator><creator>Pepperkok, Rainer</creator><creator>Huttner, Wieland B</creator><general>Nature Publishing Group US</general><general>Nature Publishing Group</general><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>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88G</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</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>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2M</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PSYQQ</scope><scope>Q9U</scope><scope>RC3</scope><scope>7QO</scope><scope>7X8</scope></search><sort><creationdate>20120201</creationdate><title>A new approach to manipulate the fate of single neural stem cells in tissue</title><author>Taverna, Elena ; Haffner, Christiane ; Pepperkok, Rainer ; Huttner, Wieland B</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c475t-8a976337cd45766ad6cee51103d8e55563106e27b7f592680abccea235a7386b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>631/136</topic><topic>631/378/2183/2182</topic><topic>631/378/2571/2579</topic><topic>Animal Genetics and Genomics</topic><topic>Animals</topic><topic>Behavioral Sciences</topic><topic>Biological Techniques</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>cdc42 GTP-Binding Protein - 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Academic</collection><jtitle>Nature neuroscience</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Taverna, Elena</au><au>Haffner, Christiane</au><au>Pepperkok, Rainer</au><au>Huttner, Wieland B</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A new approach to manipulate the fate of single neural stem cells in tissue</atitle><jtitle>Nature neuroscience</jtitle><stitle>Nat Neurosci</stitle><addtitle>Nat Neurosci</addtitle><date>2012-02-01</date><risdate>2012</risdate><volume>15</volume><issue>2</issue><spage>329</spage><epage>337</epage><pages>329-337</pages><issn>1097-6256</issn><eissn>1546-1726</eissn><coden>NANEFN</coden><abstract>This study describes a microinjection technique that allows for the acute manipulation of individual neural stem cells in organotypic slice cultures via direct delivery of biologically active molecules.
A challenge in the field of neural stem cell biology is the mechanistic dissection of single stem cell behavior in tissue. Although such behavior can be tracked by sophisticated imaging techniques, current methods of genetic manipulation do not allow researchers to change the level of a defined gene product on a truly acute time scale and are limited to very few genes at a time. To overcome these limitations, we established microinjection of neuroepithelial/radial glial cells (apical progenitors) in organotypic slice culture of embryonic mouse brain. Microinjected apical progenitors showed cell cycle parameters that were indistinguishable to apical progenitors
in utero
, underwent self-renewing divisions and generated neurons. Microinjection of single genes, recombinant proteins or complex mixtures of RNA was found to elicit acute and defined changes in apical progenitor behavior and progeny fate. Thus, apical progenitor microinjection provides a new approach to acutely manipulating single neural stem and progenitor cells in tissue.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><pmid>22179113</pmid><doi>10.1038/nn.3008</doi><tpages>9</tpages></addata></record> |
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| ispartof | Nature neuroscience, 2012-02, Vol.15 (2), p.329-337 |
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| subjects | 631/136 631/378/2183/2182 631/378/2571/2579 Animal Genetics and Genomics Animals Behavioral Sciences Biological Techniques Biomedical and Life Sciences Biomedicine cdc42 GTP-Binding Protein - administration & dosage cdc42 GTP-Binding Protein - metabolism Cell culture Cell Cycle - drug effects Cell Cycle - genetics Cell Differentiation - genetics Cell Differentiation - physiology Embryo, Mammalian Immediate-Early Proteins - genetics In Vitro Techniques Luminescent Proteins - administration & dosage Luminescent Proteins - genetics Mice Mice, Inbred C57BL Mice, Transgenic Microinjections - methods Mutation - genetics Neural Stem Cells - drug effects Neural Stem Cells - physiology Neurobiology Neurosciences Physiological aspects Rhombencephalon - cytology Rhombencephalon - embryology RNA RNA, Messenger - pharmacology Stem cells technical-report Time Factors Tumor Suppressor Proteins - genetics |
| title | A new approach to manipulate the fate of single neural stem cells in tissue |
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