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 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.
Institute of Clinical Neurobiology, University of Würzburg, Würzburg, Germany.
Beatriz
B
Blanco Redondo
Institute of Clinical Neurobiology, University of Würzburg, Würzburg, Germany.
Beatriz Blanco
BB
Redondo
Melanie
M
Bunz
Partho
P
Halder
Madhumala K
MK
Sadanandappa
Barbara
B
Mühlbauer
Felix
F
Erwin
Alois
A
Hofbauer
Veronica
V
Rodrigues
K
K
VijayRaghavan
Mani
M
Ramaswami
Dirk
D
Rieger
Christian
C
Wegener
Charlotte
C
Förster
Erich
E
Buchner
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D000941:Antigens / Q000276:immunology / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D004330:Drosophila / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006825:Hybridomas; D007150:Immunohistochemistry; D007395:Interneurons / Q000276:immunology / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence
D000911:Antibodies, Monoclonal; D000941:Antigens