Neurochemical Organization of the Drosophila Brain Visualized by Endogenously Tagged Neurotransmitter Receptors

Abstract

Highlights d Comprehensive T2A-GAL4 knockin resources for neurotransmitter receptors in flies d T2A-GAL4 can be converted into other reporters by RMCE d Quantitative analyses of receptor expression reveal segmentation of the fly brain d Four dopamine receptor proteins have distinct and dynamic subcellular localization

Neuronal octopamine signaling regulates mating-induced germline stem cell increase in female Drosophila melanogaster

“…Among these, we focused on nAChRα1 , nAChRα2, nAChRα3, nAChRβ1 and nAChRβ2 because the knock-down of these genes in dsx + Tdc2 + ( tub >GAL80>, dsx-FLP; Tdc2-GAL4 ) or Tdc2 neurons ( Tdc2-GAL4 ) increased the GSC number in virgin females similar to ppk >ChAT RNAi ( Figure 6D and Figure 6—figure supplement 1A ). We then confirmed the expression of these acetylcholine receptor genes in Tdc2 neurons by generating a knock-in T2A-GAL4 line as previously described ( Kondo et al, 2020 ; Ihara et al, 2020 ) for each 5 nAChR subunit s and observed their expression with UAS-mCD8::GFP . All of the five knock-in-GAL4 expressions were detected in anti-Tdc2 positive neurons around the ovary, suggesting that the ovary-projecting dsx + Tdc2 + neurons expresses these nAChRs ( Figure 6—figure supplement 1B–F ).…”

“…We also examined Oamb expression in the ovarian-somatic cells by two Oamb -knock-in GAL4 lines ( Deng et al, 2019 ; Kondo et al, 2020 ). However, we could not detect any reliable signals in the germarium or the mature follicle cells via these GAL4 lines with UAS-GFP and UAS-Stinger lines ( Figure 1—figure supplement 2A,B,C and D ).…”

“…The following GAL4 and LexA strains were used: c587-GAL4 ( Manseau et al, 1997 ) (gift from Hiroko Sano, Kurume University, Japan), R44E10-GAL4 ( Deady and Sun, 2015 ) (a gift from Jianjun Sun, University of Connecticut, USA) , RS-GAL4 ( Lee et al, 2009 ) (a gift from Kyung-An Han, Pennsylvania State University, USA), nSyb-GAL4 (BDSC #51941), nSyb-GAL80 ( Harris et al, 2015 ) (a gift from James W. Truman, Janelia Research Campus, USA), tj-GAL4 (DGRC #104055), R13C06-GAL4 (BDSC #47860), 109–30 GAL4 (BDSC #7023), c355-GAL4 (BDSC #3750), c306-GAL4 (BDSC #3743), slbo-GAL4 (BDSC #6458), bab1-GAL4 ( Bolívar et al, 2006 ) (a gift from Satoru Kobayashi, University of Tsukuba, Japan), nos-GAL4 (DGRC #107748), tub >FRT >GAL80>FRT (BDSC #38879), Oamb KI-RD -GAL4 (BDSC#84677) ( Deng et al, 2019 ) , Oamb-KI-T2A-GAL4, nAChRα1-T2A-GAL4, nAChRα2-T2A-GAL4, nAChRα3-T2A-GAL4, nAChRβ1-T2A-GAL4, nAChRβ2-T2A-GAL4 ( Kondo et al, 2020 ; Ihara et al, 2020 ), ChaT-GAL4 (BDSC #6793), ppk-GAL4 ( Grueber et al, 2007 ) (a gift from Hiroko Sano, Kurume University, Japan), and SPSNs-LexA ( Feng et al, 2014 ) (a gift from Young-Joon Kim, Gwangju Institute of Science and Technology, South Korea).…”

Modulation of flight and feeding behaviours requires presynaptic IP3Rs in dopaminergic neurons

“…Neurotransmitter release from PPL1-γ2α′1 (MB296B) DANs in response to stimulation of the IP 3 R was investigated next. Presence of the mAChR was confirmed on to be present on Kenyon cells and mushroom body (Kondo et al, 2020). Though, requirement for the IP 3 R and the mAChR was restricted to 48-96 hrs APF, these data suggest that the neurotransmitter acetylcholine stimulates dopamine release in adult DANs through mAChR-IP 3 R signaling.…”

Transsynaptic mapping ofDrosophilamushroom body output neurons

“…Drosophila melanogaster provides a powerful opportunity to map the fundamental architecture of neural circuits due to its complex yet tractable brain. With a nervous system of approximately 100,000 neurons and a rich genetic toolkit that offers the potential to selectively manipulate subsets of neurons in behaving animals, significant effort has been devoted to establishing a detailed map of neural connectivity in the fly (Aso et al, 2014a;Bates et al, 2020;Couto et al, 2005;Deng et al, 2019;Eichler et al, 2017;Eschbach et al, 2020;Fishilevich and Vosshall, 2005;Frechter et al, 2019;Grabe et al, 2015;Kondo et al, 2020;Li et al, 2020;Marin et al, 2020;Otto et al, 2020;Peng et al, 2011;Shao et al, 2014;Takemura et al, 2017;Zheng et al, 2018).…”