ScerGAL4LpR1-0104-G4 labels ~55 mostly dopaminergic PAM neurons. A FLP/FRT intersectional strategy shows expression in 10-20 dopaminergic neurons, innervating mushroom body beta' lobe slice 2, mushroom body gamma lobe slice 4, and TH negative neuron associated with PAM clusters that connect mushroom body gamma lobe slices 1, 2 & 4 when combined with EcollexAGMR48B04. This intersection does not label neurons innervating mushroom body gamma lobe slice 5.
Using GFP reconstitution across synaptic partners (GRASP) in a subset of dopaminergic neurons labelled by ScerGAL40104 and octopaminergic neurons labelled by ScerGAL4Tdc2.PC, reveals expression of GFP in the anterior medial protocerebrum.
Blocking the output (using shi1.UAS at 33[o]C) of Scer\GAL4LpR1-0104-G4 neurons shortly before and during training significantly impairs long-term olfactory memory in starved flies trained with sucrose, compared to controls; when Scer\GAL80R48B04 is added to this combination to reduce the the affected neurons to a subset, long-term memory is still significantly impaired in starved flies trained with sucrose and short term-memory is also significantly impaired in starved flies trained with arabinose, compared to controls.
Acute thermogenetic activation (using TrpA1UAS.(B).cKa at 33[o]C) of Scer\GAL4LpR1-0104-G4 neurons results in a significant long-term preference for a paired odor (artificial memory) in starved flies, but not flies that are fed before or after training, compared to controls.
Blocking synaptic transmission (using shi1.Scer\UAS at restrictive temperatures) from Scer\GAL4LpR1-0104-G4 neurons during acquisition significantly impairs learning in a differential aversive conditioning paradigm (in the Y[[60]] vs Z[[30]] relative condition but not the X[[0]] vs Y[[60]] absolute condition). Flies undergo conditioning at a restrictive temperature (33[o]C), whereas testing is at a permissive temperature (23[o]C); there are no significant differences when training and testing is at permissive temperatures.
