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Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the Würzburg Hybridoma Library

Several novel synaptic proteins have been identified by monoclonal antibodies (mAbs) of the Würzburg hybridoma library generated against homogenized Drosophila brains, e.g. cysteine string protein, synapse-associated protein of 47 kDa, and Bruchpilot. However, at present no routine technique exists...

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Autores principales: Redondo, Beatriz Blanco, Bunz, Melanie, Halder, Partho, Sadanandappa, Madhumala K., Mühlbauer, Barbara, Erwin, Felix, Hofbauer, Alois, Rodrigues, Veronica, VijayRaghavan, K., Ramaswami, Mani, Rieger, Dirk, Wegener, Christian, Förster, Charlotte, Buchner, Erich
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3775750/
https://www.ncbi.nlm.nih.gov/pubmed/24069413
http://dx.doi.org/10.1371/journal.pone.0075420
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author Redondo, Beatriz Blanco
Bunz, Melanie
Halder, Partho
Sadanandappa, Madhumala K.
Mühlbauer, Barbara
Erwin, Felix
Hofbauer, Alois
Rodrigues, Veronica
VijayRaghavan, K.
Ramaswami, Mani
Rieger, Dirk
Wegener, Christian
Förster, Charlotte
Buchner, Erich
author_facet Redondo, Beatriz Blanco
Bunz, Melanie
Halder, Partho
Sadanandappa, Madhumala K.
Mühlbauer, Barbara
Erwin, Felix
Hofbauer, Alois
Rodrigues, Veronica
VijayRaghavan, K.
Ramaswami, Mani
Rieger, Dirk
Wegener, Christian
Förster, Charlotte
Buchner, Erich
author_sort Redondo, Beatriz Blanco
collection PubMed
description Several novel synaptic proteins have been identified by monoclonal antibodies (mAbs) of the Würzburg hybridoma library generated against homogenized Drosophila brains, e.g. cysteine string protein, synapse-associated protein of 47 kDa, and Bruchpilot. However, at present no routine technique exists to identify the antigens of mAbs of our library that label only a small number of cells in the brain. Yet these antibodies can be used to reproducibly label and thereby identify these cells by immunohistochemical staining. Here we describe the staining patterns in the Drosophila brain for ten mAbs of the Würzburg hybridoma library. Besides revealing the neuroanatomical structure and distribution of ten different sets of cells we compare the staining patterns with those of antibodies against known antigens and GFP expression patterns driven by selected Gal4 lines employing regulatory sequences of neuronal genes. We present examples where our antibodies apparently stain the same cells in different Gal4 lines suggesting that the corresponding regulatory sequences can be exploited by the split-Gal4 technique for transgene expression exclusively in these cells. The detection of Gal4 expression in cells labeled by mAbs may also help in the identification of the antigens recognized by the antibodies which then in addition to their value for neuroanatomy will represent important tools for the characterization of the antigens. Implications and future strategies for the identification of the antigens are discussed.
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spelling pubmed-37757502013-09-25 Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the Würzburg Hybridoma Library Redondo, Beatriz Blanco Bunz, Melanie Halder, Partho Sadanandappa, Madhumala K. Mühlbauer, Barbara Erwin, Felix Hofbauer, Alois Rodrigues, Veronica VijayRaghavan, K. Ramaswami, Mani Rieger, Dirk Wegener, Christian Förster, Charlotte Buchner, Erich PLoS One Research Article Several novel synaptic proteins have been identified by monoclonal antibodies (mAbs) of the Würzburg hybridoma library generated against homogenized Drosophila brains, e.g. cysteine string protein, synapse-associated protein of 47 kDa, and Bruchpilot. However, at present no routine technique exists to identify the antigens of mAbs of our library that label only a small number of cells in the brain. Yet these antibodies can be used to reproducibly label and thereby identify these cells by immunohistochemical staining. Here we describe the staining patterns in the Drosophila brain for ten mAbs of the Würzburg hybridoma library. Besides revealing the neuroanatomical structure and distribution of ten different sets of cells we compare the staining patterns with those of antibodies against known antigens and GFP expression patterns driven by selected Gal4 lines employing regulatory sequences of neuronal genes. We present examples where our antibodies apparently stain the same cells in different Gal4 lines suggesting that the corresponding regulatory sequences can be exploited by the split-Gal4 technique for transgene expression exclusively in these cells. The detection of Gal4 expression in cells labeled by mAbs may also help in the identification of the antigens recognized by the antibodies which then in addition to their value for neuroanatomy will represent important tools for the characterization of the antigens. Implications and future strategies for the identification of the antigens are discussed. Public Library of Science 2013-09-17 /pmc/articles/PMC3775750/ /pubmed/24069413 http://dx.doi.org/10.1371/journal.pone.0075420 Text en © 2013 Blanco Redondo et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Redondo, Beatriz Blanco
Bunz, Melanie
Halder, Partho
Sadanandappa, Madhumala K.
Mühlbauer, Barbara
Erwin, Felix
Hofbauer, Alois
Rodrigues, Veronica
VijayRaghavan, K.
Ramaswami, Mani
Rieger, Dirk
Wegener, Christian
Förster, Charlotte
Buchner, Erich
Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the Würzburg Hybridoma Library
title Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the Würzburg Hybridoma Library
title_full Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the Würzburg Hybridoma Library
title_fullStr Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the Würzburg Hybridoma Library
title_full_unstemmed Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the Würzburg Hybridoma Library
title_short Identification and Structural Characterization of Interneurons of the Drosophila Brain by Monoclonal Antibodies of the Würzburg Hybridoma Library
title_sort identification and structural characterization of interneurons of the drosophila brain by monoclonal antibodies of the würzburg hybridoma library
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3775750/
https://www.ncbi.nlm.nih.gov/pubmed/24069413
http://dx.doi.org/10.1371/journal.pone.0075420
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