Tau-β-Galactosidase, an Axon-Targeted Fusion Protein
The most commonly used enzymatic reporter molecule, Escherichia coli β-galactosidase (β-gal; β-D-galactoside galactohydrolase, EC 3.2.1.23), fails to readily diffuse into axons; consequently, the morphologies of β-gal-labeled neurons cannot directly be determined. For analysis of neuronal pathfindin...
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Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1994-06, Vol.91 (13), p.5972-5976 |
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description | The most commonly used enzymatic reporter molecule, Escherichia coli β-galactosidase (β-gal; β-D-galactoside galactohydrolase, EC 3.2.1.23), fails to readily diffuse into axons; consequently, the morphologies of β-gal-labeled neurons cannot directly be determined. For analysis of neuronal pathfinding and synaptic connectivity, this information is essential. We have constructed an axon-targeted β-gal reporter by fusing the cDNA encoding the bovine microtubule-binding protein, tau, to lacZ, the E. coli gene encoding β-gal. This reporter labels cell bodies and axons when expressed by developing and adult Drosophila neurons. It also reveals the entire cellular extent of nonneuronal cells such as muscle fibers and glia. To generate neuronal markers for studies of Drosophila neural development, we constructed a tau-β-gal enhancer-trap transposon. From 1500 independent lines generated by mobilization of this transposon, we have isolated a set of useful markers for specific subsets of neurons, glia, and muscles. Since the tau cDNA-lacZ reporter utilizes bovine tau, it may also effectively target β-gal in vertebrate neurons and prove to be a useful reagent for the analysis of vertebrate nervous systems. |
doi_str_mv | 10.1073/pnas.91.13.5972 |
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For analysis of neuronal pathfinding and synaptic connectivity, this information is essential. We have constructed an axon-targeted β-gal reporter by fusing the cDNA encoding the bovine microtubule-binding protein, tau, to lacZ, the E. coli gene encoding β-gal. This reporter labels cell bodies and axons when expressed by developing and adult Drosophila neurons. It also reveals the entire cellular extent of nonneuronal cells such as muscle fibers and glia. To generate neuronal markers for studies of Drosophila neural development, we constructed a tau-β-gal enhancer-trap transposon. From 1500 independent lines generated by mobilization of this transposon, we have isolated a set of useful markers for specific subsets of neurons, glia, and muscles. Since the tau cDNA-lacZ reporter utilizes bovine tau, it may also effectively target β-gal in vertebrate neurons and prove to be a useful reagent for the analysis of vertebrate nervous systems.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.91.13.5972</identifier><identifier>PMID: 8016099</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>Animals ; Antibodies ; Axons ; Axons - physiology ; beta-Galactosidase - analysis ; beta-Galactosidase - biosynthesis ; Cattle ; Central nervous system ; Drosophila ; Drosophila - embryology ; Drosophila - physiology ; Embryo, Nonmammalian - physiology ; Embryos ; Escherichia coli - enzymology ; Immunohistochemistry ; Motor neurons ; Neuroglia ; Neurons ; Neurons - metabolism ; Neurons - physiology ; Organ Specificity ; Recombinant Fusion Proteins - analysis ; Recombinant Fusion Proteins - biosynthesis ; Restriction Mapping ; tau Proteins - analysis ; tau Proteins - biosynthesis ; Ungulates ; Vertebrates</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1994-06, Vol.91 (13), p.5972-5976</ispartof><rights>Copyright 1994 The National Academy of Sciences of the United States of America</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c559t-2b5ffe7dcec813125d52086017e9c615b792b1f0468239a0bd345bfbb943fead3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/91/13.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/2365095$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/2365095$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27901,27902,53766,53768,57992,58225</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8016099$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Callahan, Christopher A.</creatorcontrib><creatorcontrib>Thomas, John B.</creatorcontrib><title>Tau-β-Galactosidase, an Axon-Targeted Fusion Protein</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>The most commonly used enzymatic reporter molecule, Escherichia coli β-galactosidase (β-gal; β-D-galactoside galactohydrolase, EC 3.2.1.23), fails to readily diffuse into axons; consequently, the morphologies of β-gal-labeled neurons cannot directly be determined. For analysis of neuronal pathfinding and synaptic connectivity, this information is essential. We have constructed an axon-targeted β-gal reporter by fusing the cDNA encoding the bovine microtubule-binding protein, tau, to lacZ, the E. coli gene encoding β-gal. This reporter labels cell bodies and axons when expressed by developing and adult Drosophila neurons. It also reveals the entire cellular extent of nonneuronal cells such as muscle fibers and glia. To generate neuronal markers for studies of Drosophila neural development, we constructed a tau-β-gal enhancer-trap transposon. From 1500 independent lines generated by mobilization of this transposon, we have isolated a set of useful markers for specific subsets of neurons, glia, and muscles. Since the tau cDNA-lacZ reporter utilizes bovine tau, it may also effectively target β-gal in vertebrate neurons and prove to be a useful reagent for the analysis of vertebrate nervous systems.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Axons</subject><subject>Axons - physiology</subject><subject>beta-Galactosidase - analysis</subject><subject>beta-Galactosidase - biosynthesis</subject><subject>Cattle</subject><subject>Central nervous system</subject><subject>Drosophila</subject><subject>Drosophila - embryology</subject><subject>Drosophila - physiology</subject><subject>Embryo, Nonmammalian - physiology</subject><subject>Embryos</subject><subject>Escherichia coli - enzymology</subject><subject>Immunohistochemistry</subject><subject>Motor neurons</subject><subject>Neuroglia</subject><subject>Neurons</subject><subject>Neurons - metabolism</subject><subject>Neurons - physiology</subject><subject>Organ Specificity</subject><subject>Recombinant Fusion Proteins - analysis</subject><subject>Recombinant Fusion Proteins - biosynthesis</subject><subject>Restriction Mapping</subject><subject>tau Proteins - analysis</subject><subject>tau Proteins - biosynthesis</subject><subject>Ungulates</subject><subject>Vertebrates</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1OGzEUha2qKA3QdTdtlRVsmORee_wndYMQfxISLMLa8sx46KDJOLU9FbwWD8IzMVFCaDew8uJ85-haHyHfEKYIks2WnY1TjVNkU64l_UTGCBozkWv4TMYAVGYqp_kXshvjPQBormBERgpQgNZjwue2z56fsnPb2jL52FQ2uqOJ7SbHD77L5jbcueSqyVkfG99NboJPrun2yU5t2-i-bt49cnt2Oj-5yK6uzy9Pjq-yknOdMlrwunayKl2pkCHlFaegBKB0uhTIC6lpgTXkQlGmLRQVy3lRF4XOWe1sxfbIr_Xusi8WbtjpUrCtWYZmYcOj8bYx_ydd89vc-b8mzxH1UD_Y1IP_07uYzKKJpWtb2znfRyMFl6hQfQii4EJJQQdwtgbL4GMMrt7egmBWQsxKiNFokJmVkKHx498vbPmNgSE_3OSr4mv6NmDqvm2Te0gD-fNdcgC-r4H7mHzYEpQJPnhnL7ymqRc</recordid><startdate>19940621</startdate><enddate>19940621</enddate><creator>Callahan, Christopher A.</creator><creator>Thomas, John B.</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</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>7SS</scope><scope>7TK</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19940621</creationdate><title>Tau-β-Galactosidase, an Axon-Targeted Fusion Protein</title><author>Callahan, Christopher A. ; Thomas, John B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c559t-2b5ffe7dcec813125d52086017e9c615b792b1f0468239a0bd345bfbb943fead3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Axons</topic><topic>Axons - physiology</topic><topic>beta-Galactosidase - analysis</topic><topic>beta-Galactosidase - biosynthesis</topic><topic>Cattle</topic><topic>Central nervous system</topic><topic>Drosophila</topic><topic>Drosophila - embryology</topic><topic>Drosophila - physiology</topic><topic>Embryo, Nonmammalian - physiology</topic><topic>Embryos</topic><topic>Escherichia coli - enzymology</topic><topic>Immunohistochemistry</topic><topic>Motor neurons</topic><topic>Neuroglia</topic><topic>Neurons</topic><topic>Neurons - metabolism</topic><topic>Neurons - physiology</topic><topic>Organ Specificity</topic><topic>Recombinant Fusion Proteins - analysis</topic><topic>Recombinant Fusion Proteins - biosynthesis</topic><topic>Restriction Mapping</topic><topic>tau Proteins - analysis</topic><topic>tau Proteins - biosynthesis</topic><topic>Ungulates</topic><topic>Vertebrates</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Callahan, Christopher A.</creatorcontrib><creatorcontrib>Thomas, John B.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Neurosciences Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Callahan, Christopher A.</au><au>Thomas, John B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Tau-β-Galactosidase, an Axon-Targeted Fusion Protein</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1994-06-21</date><risdate>1994</risdate><volume>91</volume><issue>13</issue><spage>5972</spage><epage>5976</epage><pages>5972-5976</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>The most commonly used enzymatic reporter molecule, Escherichia coli β-galactosidase (β-gal; β-D-galactoside galactohydrolase, EC 3.2.1.23), fails to readily diffuse into axons; consequently, the morphologies of β-gal-labeled neurons cannot directly be determined. For analysis of neuronal pathfinding and synaptic connectivity, this information is essential. We have constructed an axon-targeted β-gal reporter by fusing the cDNA encoding the bovine microtubule-binding protein, tau, to lacZ, the E. coli gene encoding β-gal. This reporter labels cell bodies and axons when expressed by developing and adult Drosophila neurons. It also reveals the entire cellular extent of nonneuronal cells such as muscle fibers and glia. To generate neuronal markers for studies of Drosophila neural development, we constructed a tau-β-gal enhancer-trap transposon. From 1500 independent lines generated by mobilization of this transposon, we have isolated a set of useful markers for specific subsets of neurons, glia, and muscles. Since the tau cDNA-lacZ reporter utilizes bovine tau, it may also effectively target β-gal in vertebrate neurons and prove to be a useful reagent for the analysis of vertebrate nervous systems.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>8016099</pmid><doi>10.1073/pnas.91.13.5972</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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source | Jstor Complete Legacy; MEDLINE; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
subjects | Animals Antibodies Axons Axons - physiology beta-Galactosidase - analysis beta-Galactosidase - biosynthesis Cattle Central nervous system Drosophila Drosophila - embryology Drosophila - physiology Embryo, Nonmammalian - physiology Embryos Escherichia coli - enzymology Immunohistochemistry Motor neurons Neuroglia Neurons Neurons - metabolism Neurons - physiology Organ Specificity Recombinant Fusion Proteins - analysis Recombinant Fusion Proteins - biosynthesis Restriction Mapping tau Proteins - analysis tau Proteins - biosynthesis Ungulates Vertebrates |
title | Tau-β-Galactosidase, an Axon-Targeted Fusion Protein |
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