Flight and Climbing Assay for Assessing Motor Functions in Drosophila

Manjila, Steffy B.; Hasan, Gaiti

doi:10.21769/bioprotoc.2742

Cited by 35 publications

(22 citation statements)

References 3 publications

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“…Subsequent studies identified mild flight deficits upon RNAi mediated knock down of the IP 3 R in dopaminergic neurons (DANs) ( Pathak et al, 2015 ) when tethered flight was tested for 30 s. Knock-down of the IP 3 R in serotonergic neurons, that form another major subset of the tested monoaminergic neurons, did not give a flight deficit ( Sadaf et al, 2012 ). Here we tested if pan-neuronal expression of the IP 3 R DN affects longer flight bout durations in a modified tethered flight assay lasting for 15 min (see Materials and methods and Manjila and Hasan, 2018 ). Flies with pan-neuronal nsybGAL4 -driven expression of Itpr DN exhibit significantly reduced flight bouts (281.4 ± 38.9 s) as compared to the appropriate genetic controls Itpr DN /+ (773.3 ± 30.4 s) and nsyb/+ (670.6 ± 41.3 s) ( Figure 2—figure supplement 1A , Figure 2—source data 4 ).…”

Section: Resultsmentioning

confidence: 99%

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

Sharma

Hasan

2020

eLife

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Innate behaviours, although robust and hard wired, rely on modulation of neuronal circuits, for eliciting an appropriate response according to internal states and external cues. Drosophila flight is one such innate behaviour that is modulated by intracellular calcium release through inositol 1,4,5-trisphosphate receptors (IP3Rs). Cellular mechanism(s) by which IP3Rs modulate neuronal function for specific behaviours remain speculative, in vertebrates and invertebrates. To address this, we generated an inducible dominant negative form of the IP3R (IP3RDN). Flies with neuronal expression of IP3RDN exhibit flight deficits. Expression of IP3RDN helped identify key flight-modulating dopaminergic neurons with axonal projections in the mushroom body. Flies with attenuated IP3Rs in these presynaptic dopaminergic neurons exhibit shortened flight bouts and a disinterest in seeking food, accompanied by reduced excitability and dopamine release upon cholinergic stimulation. Our findings suggest that the same neural circuit modulates the drive for food search and for undertaking longer flight bouts.

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Section: Resultsmentioning

confidence: 99%

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

Sharma

Hasan

2020

eLife

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“…Single Flight Assay Flies of either sex were aged for 3-5 days and tested for flight as explained in detail [55]. Briefly, flies were anaesthetized on ice for a short period ($5min) and tethered between their head and thorax using a thin metal wire and nail polish.…”

Section: Methods Detailsmentioning

confidence: 99%

Extended Flight Bouts Require Disinhibition from GABAergic Mushroom Body Neurons

et al. 2019

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“…Rapid induced negative geotaxis (Liu et al, 2015;Cao et al, 2017;Manjila and Hasan, 2018) was used as a simple test for sensorymotor performance. Experiments were done in the late afternoon.…”

Section: Climbing Assaymentioning

confidence: 99%

“…This assay was performed as described in detail previously (Agrawal et al, 2013;Manjila and Hasan, 2018). Briefly, flies were anesthetized for 10-15 min on ice and then attached with nail polish between head and thorax to a thin wire.…”

Section: Flight Duration Assaymentioning

confidence: 99%

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Implications of the Sap47 null mutation for synapsin phosphorylation, longevity, climbing, and behavioural plasticity in adult Drosophila

Blanco-Redondo

Nuwal

Kneitz

et al. 2019

Journal of Experimental Biology

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The Sap47 gene of Drosophila melanogaster encodes a highly abundant 47 kDa synaptic vesicle-associated protein. Sap47 null mutants show defects in synaptic plasticity and larval olfactory associative learning but the molecular function of Sap47 at the synapse is unknown. We demonstrate that Sap47 modulates the phosphorylation of another highly abundant conserved presynaptic protein, synapsin. Site-specific phosphorylation of Drosophila synapsin has repeatedly been shown to be important for behavioural plasticity but it was not known where these phospho-synapsin isoforms are localized in the brain. Here, we report the distribution of serine-6-phosphorylated synapsin in the adult brain and show that it is highly enriched in rings of synapses in the ellipsoid body and in large synapses near the lateral triangle. The effects of knockout of Sap47 or synapsin on olfactory associative learning/memory support the hypothesis that both proteins operate in the same molecular pathway. We therefore asked if this might also be true for other aspects of their function. We show that knockout of Sap47 but not synapsin reduces lifespan, whereas knockout of Sap47 and synapsin, either individually or together, affects climbing proficiency, as well as plasticity in circadian rhythms and sleep. Furthermore, electrophysiological assessment of synaptic properties at the larval neuromuscular junction (NMJ) reveals increased spontaneous synaptic vesicle fusion and reduced paired pulse facilitation in Sap47 and synapsin single and double mutants. Our results imply that Sap47 and synapsin cooperate non-uniformly in the control of synaptic properties in different behaviourally relevant neuronal networks of the fruitfly.

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Flight and Climbing Assay for Assessing Motor Functions in Drosophila

Cited by 35 publications

References 3 publications

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

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

Extended Flight Bouts Require Disinhibition from GABAergic Mushroom Body Neurons

Implications of the Sap47 null mutation for synapsin phosphorylation, longevity, climbing, and behavioural plasticity in adult Drosophila

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