Identification of Neuronal Subtypes in Drosophila Using Genetic Tools

We used wild type (Oregon R) and white eyed (w¹¹¹⁸) fruitflies, Drosophila melanogaster for immunocytochemistry and in situ hybridization. A number of Gal4 strains were used for driving GFP to identify specific sets of neurons in relation to sNPF markers. These were: gad1-Gal4 (Glutamic acid decarboxylase-1 promoter-Gal4 [81 (link)]; from G. Miesenböck, New Haven, CT), Cha-Gal4 (w*; P{Cha-GAL4.7.4}19B P{UAS-GFP.S65T}T2; choline acetyltransferase promotor-Gal4-GFP fusion, from Bloomington Drosophila Stock Center at Indiana University, IN [33 (link)]); th-Gal4 (tyrosine-hydroxylase promoter-Gal4; [38 (link)]; from S. Birmann, Marseille, France), tdc-Gal4 (tyrosine decarboxylase promoter Gal4; obtained from J. Dubnau, Cold Spring Harbor, NY; [82 (link)]), OK371-Gal4 (vesicular glutamate transporter-Gal4) (from H. Aberle, Tübingen, Germany [36 (link)]; OK107-Gal4 (P{GawB}OK107 (expression in mushroom body neurons); from Bloomington Stock center; [83 (link),84 (link)]), npf-Gal4 (Neuropeptide F-promoter-Gal4; from P. Shen, Athens, GA; [39 (link)]). A snpf-Gal4 (NP6301; order number 113901) was obtained from the Drosophila Genetic Resource Center (DGRC), Kyoto Institute of Technology, Kyoto, Japan. The genotype of this Gal4 is yw; P{GawB}NP6301/CyO, P{UAS-lacZ.UW14}UW14. To our knowledge the expression of this Gal4 has not been previously described. The enhancer trap line Mai179-Gal4 [27 (link)] was obtained from G. Korge (Berlin, Germany). This drives expression in a subset of neurons in the brain including some neurosecretory cells. The c929-Gal4 line was obtained from P. H. Taghert (St. Louis, MO). This Gal4 reveals the Drosophila dimmed (dimm) gene that encodes a bHLH protein, DIMM, known to be required for the differentiation of peptidergic neurons and endocrine cells [24 (link)]. Finally, UAS-cd8gfp flies (from Bloomington Stock center) were used as targets to express GFP.

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Nässel D.R., Enell L.E., Santos J.G., Wegener C, & Johard H.A. (2008). A large population of diverse neurons in the Drosophila central nervous system expresses short neuropeptide F, suggesting multiple distributed peptide functions. BMC Neuroscience, 9, 90.

Publication 2008

Bhlh protein Brain Cells Choline acetyltransferase Cold Drosophila Drosophila melanogaster Endocrine cells Flies Gad1 Gene Genotype Glutamic acid decarboxylase Immunocytochemistry In situ hybridization Lacz Mushroom body Neurons Neuropeptide f Strains Tyrosine decarboxylase Tyrosine hydroxylase Vesicular glutamate transporter

Corresponding Organization :

Other organizations : Stockholm University, Philipps University of Marburg

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Variable analysis

independent variables

Wild type (Oregon R) and white eyed (w^1118) fruitflies, Drosophila melanogaster
Gal4 strains used for driving GFP to identify specific sets of neurons in relation to sNPF markers: gad1-Gal4, Cha-Gal4, th-Gal4, tdc-Gal4, OK371-Gal4, OK107-Gal4, npf-Gal4, snpf-Gal4, Mai179-Gal4, c929-Gal4
UAS-cd8gfp flies used as targets to express GFP

dependent variables

Immunocytochemistry and in situ hybridization

control variables

White eyed (w^1118) fruitflies
Gal4 strains used as controls to identify specific sets of neurons in relation to sNPF markers

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