A small cohort of FRUM and Engrailed-expressing neurons mediate successful copulation in Drosophila melanogaster
1 Department of Zoology, Molecular and Cellular Biology Program, Oregon State University, Corvallis, Oregon 97331-2914, USA
2 Molecular and Cellular Biology Program, Oregon State University, Corvallis, Oregon 97361, USA
3 Present address: Biology Department, Western Oregon University, Monmouth, Oregon 97361, USA
4 Present address: National Health Research Institutes, Zhunan, Miaoli 35053, Taiwan
BMC Neuroscience 2013, 14:57 doi:10.1186/1471-2202-14-57Published: 21 May 2013
Additional file 1: Table S1:
En-fruMRNAi males have normal courtship index (CI) values. Measurements from 10-minute videotaped courtship tests (see Methods) include courtship index (a measure of time spent performing wing courtship song). All genotypes were not statistically different for courtship index (One-Way ANOVA, p = 0.019).
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Additional file 2: Figure S1:
En-fruMRNAi males make and store sperm and mating plug material. In dissected reproductive tracts from wild-type (A, B) and En-fruMRNAi (C, D) males, sperm was viewed by differential interference contrast microscopy and mating plug material was visible under ultraviolet light. Sperm and mating plug material levels appeared to be normal, and sperm were motile.
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Additional file 3: Figure S2:
En-fruMRNAi males have normal serotonergic innervation. Serotonergic nerve terminals innervating the internal reproductive organs were examined in wild-type and en-GAL4/UAS-fruMIR males by immunohistochemistry with anti-serotonin (5HT). A) In a wild-type male, serotonergic nerve terminals are present on the seminal vesicles (sv), accessory glands (ag) and ejaculatory duct (ed). B) In an En-fruMRNAi male, serotonergic terminals are present on the same organs, similar to wild-type males. Images are confocal z-stacks through the male internal reproductive tract. Size bar = 200 um.
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Additional file 4: Figure S3:
Wild-type males, females, and fru mutant males have similar neurotransmitter profiles for brain and VNC-T1midline neurons, but vary for VNC-medial neurons in T1, T2, and T3. CNSs of wild-type males (A, C, F, G, H), wild-type females (B, D, I), and fru-mutant males (E, J) were labeled for anti-FRUM (green) and anti-Engrailed (magenta), and a neurochemical marker (blue). (A, B) E/F-VNCmid neurons in males, and the equivalent in females, express gamma-Aminobutyric acid (GABA) neurotransmitter as labeled by anti-GABA antibody. (C, D, E) In males, females, and fru mutant males E/F-VNCmid neurons are also labeled by anti-GAD antibody (GAD = Glutamic Acid Decarboxylase, an enzyme for GABA synthesis, localized exclusively to GABAergic neurons). (F, G) E/F-brain (F) and E/F-AbG (G) neurons are also labeled by GAD in wild-type males. (H, I, J) Males expressed a Ddc-GAL4; UAS-mcd8::GFP in E/F-VNCmed neurons of T1, T2 (H) and T3 (not shown). Females (I) and fru-mutant males (J) express this driver much more faintly, in with variable penetrance in the equivalent neurons. (Ddc = dopa Decarboxylase, an enzyme for serotonin/5HTsynthesis). (K, L, M) Schematic indicating neurotransmitter profile of En/FRUM neurons in the brain (K), T1/T2 segments of the VNC (L), and T3 and AbG (M).
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