The notch gene product is a glycoprotein expressed on the cell surface of both epidermal and neuronal precursor cells during Drosophila development
The Notch locus of Drosophila melanogaster is one of a small number of zygotically acting "neurogenic" genes involved in the correct segregation of neural from epidermal lineages during embryogenesis as well as in other postembryonic developmental events. We have generated antibody probes...
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Veröffentlicht in: | The Journal of cell biology 1989-11, Vol.109 (5), p.2427-2440 |
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description | The Notch locus of Drosophila melanogaster is one of a small number of zygotically acting "neurogenic" genes involved in the correct segregation of neural from epidermal lineages during embryogenesis as well as in other postembryonic developmental events. We have generated antibody probes against three regions of the Notch protein to study the expression of Notch and begin a biochemical characterization of the protein. Consistent with predictions based on DNA sequence data, here we gather evidence showing that Notch encodes a large, glycosylated surface protein with an apparent molecular mass of 300 kD: (a) all three antibodies detect Notch on Western blots as a high molecular mass, primarily full-length product; (b) immunoelectron microscopy localizes the Notch protein to the cell membrane; and (c) lentil lectin column binding demonstrates that the protein is glycosylated, indicative of its surface protein nature. In general, the distribution of the Notch protein coincides with that of the Notch transcript determined previously by in situ hybridizations. Notch is expressed in a much wider range of tissue types than those disrupted in the neurogenic mutant, as determined by antibody localization. Early labeling in the blastoderm appears ubiquitous except for the pole cells, but as development proceeds some distinctive features emerge: stronger staining is seen within the germ band layer where neuroblast delamination occurs, and the developing embryonic nervous system shows pronounced axonal staining. In third instar larvae, Notch is expressed in imaginal disks and in the central nervous system. Based on these results, certain models for how Notch controls the neuroblast cell fate choice are eliminated. We discuss how Notch may function in this choice as well as in other lineage fate determinations. |
doi_str_mv | 10.1083/jcb.109.5.2427 |
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(Iowa State University, Ames, IA) ; Fehon, R.G ; Artavanis-Tsakonas, S</creator><creatorcontrib>Johansen, K.M. (Iowa State University, Ames, IA) ; Fehon, R.G ; Artavanis-Tsakonas, S</creatorcontrib><description>The Notch locus of Drosophila melanogaster is one of a small number of zygotically acting "neurogenic" genes involved in the correct segregation of neural from epidermal lineages during embryogenesis as well as in other postembryonic developmental events. We have generated antibody probes against three regions of the Notch protein to study the expression of Notch and begin a biochemical characterization of the protein. Consistent with predictions based on DNA sequence data, here we gather evidence showing that Notch encodes a large, glycosylated surface protein with an apparent molecular mass of 300 kD: (a) all three antibodies detect Notch on Western blots as a high molecular mass, primarily full-length product; (b) immunoelectron microscopy localizes the Notch protein to the cell membrane; and (c) lentil lectin column binding demonstrates that the protein is glycosylated, indicative of its surface protein nature. In general, the distribution of the Notch protein coincides with that of the Notch transcript determined previously by in situ hybridizations. Notch is expressed in a much wider range of tissue types than those disrupted in the neurogenic mutant, as determined by antibody localization. Early labeling in the blastoderm appears ubiquitous except for the pole cells, but as development proceeds some distinctive features emerge: stronger staining is seen within the germ band layer where neuroblast delamination occurs, and the developing embryonic nervous system shows pronounced axonal staining. In third instar larvae, Notch is expressed in imaginal disks and in the central nervous system. Based on these results, certain models for how Notch controls the neuroblast cell fate choice are eliminated. We discuss how Notch may function in this choice as well as in other lineage fate determinations.</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.109.5.2427</identifier><identifier>PMID: 2509485</identifier><identifier>CODEN: JCLBA3</identifier><language>eng</language><publisher>New York, NY: Rockefeller University Press</publisher><subject>ANIMAL DEVELOPMENTAL STAGES ; Animals ; Antibodies ; Biological and medical sciences ; Blotting, Western ; Cell lines ; Cell Membrane - metabolism ; Cells ; Central nervous system ; Cloning, Molecular ; Diptera ; DNA - genetics ; DROSOPHILA ; DROSOPHILA MELANOGASTER ; Drosophila melanogaster - genetics ; Drosophila melanogaster - growth & development ; Drosophilidae ; Embryos ; Epidermal Cells ; Epidermis - metabolism ; ETAPAS DEL DESARROLLO ANIMAL ; Fundamental and applied biological sciences. Psychology ; GENE ; Gene expression ; GENES ; Genetic loci ; GLICOPROTEINAS ; GLYCOPROTEINE ; GLYCOPROTEINS ; Larva ; Membrane Glycoproteins - genetics ; Molecular and cellular biology ; Molecular genetics ; Nerve Tissue Proteins - genetics ; Neurons ; Neurons - cytology ; Neurons - metabolism ; Notch receptors ; Pupa ; Repetitive Sequences, Nucleic Acid ; Restriction Mapping ; STADE DE DEVELOPPEMENT ANIMAL</subject><ispartof>The Journal of cell biology, 1989-11, Vol.109 (5), p.2427-2440</ispartof><rights>Copyright 1989 The Rockefeller University Press</rights><rights>1990 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c551t-7189dbfd347aa800978190a8c76bb2e0728349b656d06859d2f5acddb1c14a9d3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2115861/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2115861/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,728,781,785,886,27929,27930,53796,53798</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6655428$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2509485$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Johansen, K.M. (Iowa State University, Ames, IA)</creatorcontrib><creatorcontrib>Fehon, R.G</creatorcontrib><creatorcontrib>Artavanis-Tsakonas, S</creatorcontrib><title>The notch gene product is a glycoprotein expressed on the cell surface of both epidermal and neuronal precursor cells during Drosophila development</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>The Notch locus of Drosophila melanogaster is one of a small number of zygotically acting "neurogenic" genes involved in the correct segregation of neural from epidermal lineages during embryogenesis as well as in other postembryonic developmental events. We have generated antibody probes against three regions of the Notch protein to study the expression of Notch and begin a biochemical characterization of the protein. Consistent with predictions based on DNA sequence data, here we gather evidence showing that Notch encodes a large, glycosylated surface protein with an apparent molecular mass of 300 kD: (a) all three antibodies detect Notch on Western blots as a high molecular mass, primarily full-length product; (b) immunoelectron microscopy localizes the Notch protein to the cell membrane; and (c) lentil lectin column binding demonstrates that the protein is glycosylated, indicative of its surface protein nature. In general, the distribution of the Notch protein coincides with that of the Notch transcript determined previously by in situ hybridizations. Notch is expressed in a much wider range of tissue types than those disrupted in the neurogenic mutant, as determined by antibody localization. Early labeling in the blastoderm appears ubiquitous except for the pole cells, but as development proceeds some distinctive features emerge: stronger staining is seen within the germ band layer where neuroblast delamination occurs, and the developing embryonic nervous system shows pronounced axonal staining. In third instar larvae, Notch is expressed in imaginal disks and in the central nervous system. Based on these results, certain models for how Notch controls the neuroblast cell fate choice are eliminated. We discuss how Notch may function in this choice as well as in other lineage fate determinations.</description><subject>ANIMAL DEVELOPMENTAL STAGES</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>Cell lines</subject><subject>Cell Membrane - metabolism</subject><subject>Cells</subject><subject>Central nervous system</subject><subject>Cloning, Molecular</subject><subject>Diptera</subject><subject>DNA - genetics</subject><subject>DROSOPHILA</subject><subject>DROSOPHILA MELANOGASTER</subject><subject>Drosophila melanogaster - genetics</subject><subject>Drosophila melanogaster - growth & development</subject><subject>Drosophilidae</subject><subject>Embryos</subject><subject>Epidermal Cells</subject><subject>Epidermis - metabolism</subject><subject>ETAPAS DEL DESARROLLO ANIMAL</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>GENE</subject><subject>Gene expression</subject><subject>GENES</subject><subject>Genetic loci</subject><subject>GLICOPROTEINAS</subject><subject>GLYCOPROTEINE</subject><subject>GLYCOPROTEINS</subject><subject>Larva</subject><subject>Membrane Glycoproteins - genetics</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Nerve Tissue Proteins - genetics</subject><subject>Neurons</subject><subject>Neurons - cytology</subject><subject>Neurons - metabolism</subject><subject>Notch receptors</subject><subject>Pupa</subject><subject>Repetitive Sequences, Nucleic Acid</subject><subject>Restriction Mapping</subject><subject>STADE DE DEVELOPPEMENT ANIMAL</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFUc2P1CAUJ0azjqNXD0YTDsZbR2ihlMsmZv1MNvHg7plQeG2ZtFCh3bh_h_-wjDMZ9eSJB7-Px3s_hJ5TsqOkqd7uTZsLueO7kpXiAdpQzkjRUEYeog0hJS0kL_lj9CSlPSGECVZdoIuSE8kavkE_bwbAPixmwD14wHMMdjULdglr3I_3JuSXBZzH8GOOkBJYHDxessrAOOK0xk4bwKHDbVgGDLOzECc9Yu0t9rDG4PMlS80aU4i_VQnbNTrf4_cxpDAPbtTYwh2MYZ7AL0_Ro06PCZ6dzi26_fjh5upzcf3105erd9eF4ZwuhaCNtG1nKya0bgiRoqGS6MaIum1LIKJsKibbmteW1A2Xtuy4Nta21FCmpa226PLoO6_tBNbk1lGPao5u0vFeBe3Uv4h3g-rDnSop5U1Ns8Gbk0EM31dIi5pcOgyoPYQ1KSFLSSkT_yXmzHJgss7E3ZFo8mZShO78G0rUIW-V886FVFwd8s6CV3_PcKafAs746xOuk9FjF7U3Lp1pdc05y3vaopdH2j4tIf5pmocUjGT4xRHudFC6j9nh9pskVFaCVr8Abl3I0A</recordid><startdate>19891101</startdate><enddate>19891101</enddate><creator>Johansen, K.M. (Iowa State University, Ames, IA)</creator><creator>Fehon, R.G</creator><creator>Artavanis-Tsakonas, S</creator><general>Rockefeller University Press</general><general>The Rockefeller University Press</general><scope>FBQ</scope><scope>IQODW</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>7SS</scope><scope>7TK</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19891101</creationdate><title>The notch gene product is a glycoprotein expressed on the cell surface of both epidermal and neuronal precursor cells during Drosophila development</title><author>Johansen, K.M. (Iowa State University, Ames, IA) ; Fehon, R.G ; Artavanis-Tsakonas, S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c551t-7189dbfd347aa800978190a8c76bb2e0728349b656d06859d2f5acddb1c14a9d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>ANIMAL DEVELOPMENTAL STAGES</topic><topic>Animals</topic><topic>Antibodies</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>Cell lines</topic><topic>Cell Membrane - metabolism</topic><topic>Cells</topic><topic>Central nervous system</topic><topic>Cloning, Molecular</topic><topic>Diptera</topic><topic>DNA - genetics</topic><topic>DROSOPHILA</topic><topic>DROSOPHILA MELANOGASTER</topic><topic>Drosophila melanogaster - genetics</topic><topic>Drosophila melanogaster - growth & development</topic><topic>Drosophilidae</topic><topic>Embryos</topic><topic>Epidermal Cells</topic><topic>Epidermis - metabolism</topic><topic>ETAPAS DEL DESARROLLO ANIMAL</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>GENE</topic><topic>Gene expression</topic><topic>GENES</topic><topic>Genetic loci</topic><topic>GLICOPROTEINAS</topic><topic>GLYCOPROTEINE</topic><topic>GLYCOPROTEINS</topic><topic>Larva</topic><topic>Membrane Glycoproteins - genetics</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Nerve Tissue Proteins - genetics</topic><topic>Neurons</topic><topic>Neurons - cytology</topic><topic>Neurons - metabolism</topic><topic>Notch receptors</topic><topic>Pupa</topic><topic>Repetitive Sequences, Nucleic Acid</topic><topic>Restriction Mapping</topic><topic>STADE DE DEVELOPPEMENT ANIMAL</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Johansen, K.M. (Iowa State University, Ames, IA)</creatorcontrib><creatorcontrib>Fehon, R.G</creatorcontrib><creatorcontrib>Artavanis-Tsakonas, S</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><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>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Johansen, K.M. (Iowa State University, Ames, IA)</au><au>Fehon, R.G</au><au>Artavanis-Tsakonas, S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The notch gene product is a glycoprotein expressed on the cell surface of both epidermal and neuronal precursor cells during Drosophila development</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>1989-11-01</date><risdate>1989</risdate><volume>109</volume><issue>5</issue><spage>2427</spage><epage>2440</epage><pages>2427-2440</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><coden>JCLBA3</coden><abstract>The Notch locus of Drosophila melanogaster is one of a small number of zygotically acting "neurogenic" genes involved in the correct segregation of neural from epidermal lineages during embryogenesis as well as in other postembryonic developmental events. We have generated antibody probes against three regions of the Notch protein to study the expression of Notch and begin a biochemical characterization of the protein. Consistent with predictions based on DNA sequence data, here we gather evidence showing that Notch encodes a large, glycosylated surface protein with an apparent molecular mass of 300 kD: (a) all three antibodies detect Notch on Western blots as a high molecular mass, primarily full-length product; (b) immunoelectron microscopy localizes the Notch protein to the cell membrane; and (c) lentil lectin column binding demonstrates that the protein is glycosylated, indicative of its surface protein nature. In general, the distribution of the Notch protein coincides with that of the Notch transcript determined previously by in situ hybridizations. Notch is expressed in a much wider range of tissue types than those disrupted in the neurogenic mutant, as determined by antibody localization. Early labeling in the blastoderm appears ubiquitous except for the pole cells, but as development proceeds some distinctive features emerge: stronger staining is seen within the germ band layer where neuroblast delamination occurs, and the developing embryonic nervous system shows pronounced axonal staining. In third instar larvae, Notch is expressed in imaginal disks and in the central nervous system. Based on these results, certain models for how Notch controls the neuroblast cell fate choice are eliminated. We discuss how Notch may function in this choice as well as in other lineage fate determinations.</abstract><cop>New York, NY</cop><pub>Rockefeller University Press</pub><pmid>2509485</pmid><doi>10.1083/jcb.109.5.2427</doi><tpages>14</tpages><oa>free_for_read</oa></addata></record> |
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subjects | ANIMAL DEVELOPMENTAL STAGES Animals Antibodies Biological and medical sciences Blotting, Western Cell lines Cell Membrane - metabolism Cells Central nervous system Cloning, Molecular Diptera DNA - genetics DROSOPHILA DROSOPHILA MELANOGASTER Drosophila melanogaster - genetics Drosophila melanogaster - growth & development Drosophilidae Embryos Epidermal Cells Epidermis - metabolism ETAPAS DEL DESARROLLO ANIMAL Fundamental and applied biological sciences. Psychology GENE Gene expression GENES Genetic loci GLICOPROTEINAS GLYCOPROTEINE GLYCOPROTEINS Larva Membrane Glycoproteins - genetics Molecular and cellular biology Molecular genetics Nerve Tissue Proteins - genetics Neurons Neurons - cytology Neurons - metabolism Notch receptors Pupa Repetitive Sequences, Nucleic Acid Restriction Mapping STADE DE DEVELOPPEMENT ANIMAL |
title | The notch gene product is a glycoprotein expressed on the cell surface of both epidermal and neuronal precursor cells during Drosophila development |
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