Choline Transporter in α/β core neurons of Drosophila mushroom body non-canonically regulates pupal eclosion and maintains neuromuscular junction integrity

Abstract

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Choline Transporter regulates olfactory habituation via a neuronal triad of excitatory, inhibitory and mushroom body neurons

“…NP1131 (103898) was from DGRC Kyoto stock centre, Japan. UAS-ChT line was previously created and characterised (Hamid et al, 2019). All genetic combinations of stocks used in this study were created in the lab.…”

“…To further ascertain that attenuation of naïve chemotactic response and habituation index is indeed due to knockdown of ChT and not due to non-specific secondary effect of the transgenic lines, we also examined the heterozygous combination of UAS-ChT RNAi1 and UAS-ChT RNAi2 with wildtype lines (UAS-ChT RNAi1 >+ or UAS-ChT RNAi2 >+) and did not observe any significantly different response compared to the different MBGAL4s>+.(Fig. S5).Previously, we reported attenuated neuromuscular junctions (NMJ) in third instar larvae caused due to knockdown of ChT in α/β and ϒ intrinsic lobes of MB(Hamid et al, 2019). To ascertain if the observed reduction in habituation index is the acute functional change in the neurons during habituation or have resulted due to changes in the NMJ, we employed the TARGET system (McGuire et al, 2004).…”

“…The MB in Drosophila brain are bilateral neuropilar areas which are principally cholinergic expressing ChAT and VAChT (Barnstedt et al, 2016;Croset et al, 2018). Based on immunostainings, we recently reported that there is a preponderance of ChT in Drosophila MB as compared to ChAT and VAChT (Hamid et al, 2019). The functional relevance of ChT expression in MB is not clear.…”

“…The knockdown efficiency of ChT RNAi2 was assessed by immunostaining (Fig. S1) while of ChT RNAi1 was described previously by us(Hamid et al, 2019). First, we assessed the naïve chemotactic response of 3 rd instar foraging larvae towards ETA upon knockdown of ChT in each of the neuronal domain of MB.…”