A ple genomic fragment from -4168 to +136bp relative to the transcription start site is fused upstream of sequence encoding a GAL4 driver plus a 3' translation termination element from the Hsp70 gene. The ple coding sequence (from +183 to +6820 relative to the transcription start site) is fused downstream of the Hsp70 translation termination sequence (so that any regulatory sequences present in the coding region are included, but the ple coding sequence is not translated).
Drives expression in dopaminergic neurons.
Expression driven by Scer\GAL4ple.PF was restricted via a transgenic cell class-lineage intersection (CLIn) system to a subset of neuronal clusters - PPM3 and a subset of PPM2 and PPL1 neurons - of specific Type II neuroblast lineages using Klac\KDRstg14. All 8 PPM3 neurons are part of the DM6 neuroblast lineage, two of these, originating from the first larval born secondary neuroblast, are EB neurons, while the mFB and vFB neurons originate from other secondary neuroblasts. The subset of PPM2 neurons are part of the DM2 (0-1 neurons), DM3 (0-1 neurons) and DM4 (1 neuron) neuroblast lineages. The 2 dFB neurons of the dopaminergic PPL1 cluster are part of the DL1 neuroblast lineage, while the remaining 5 non-dFB neurons are part of the DL2 neuroblast lineage. These 5 neurons include a MB-alpha neuron of the dopaminergic PPL1 cluster, a MB-alpha' neuron of the dopaminergic PPL1 cluster, a mushroom body vertical lobe arborizing neuron 1 and a further neuron that targets the superior medial protocerebrum and superior intermediate protocerebrum.
Scer\GAL4ple.PF drives expression in 1-3 SP1 neurons, 1-4 SP2 neurons, 3-6 IP neurons, all LP1 neurons, 2-4 LP2 neurons, 1-3 SE1 neurons, 1-3 adult serotonergic LP3 neurons and 1 adult serotonergic AP neuron. Additionally, it also drives expression in small numbers (1-14) of non-serotonergic neurons adjacent to SP1, SP2, IP, LP1, LP2, LP3, SE1, SE3, T1, T2, T3, and abdominal serotonergic neurons.
Scer\GAL4ple.PF drives expression in all dopaminergic neurons of the PAL PPM1/2, PPL1. PPM3 and PPL2 clusters which project to the vertical lobes of the mushroom body, the ellipsoid and the fan-shaped body. Expression is only found in around 10% of dopaminergic neurons of the PAM cluster.
Scer\GAL4ple.PF drives expression in the dopaminergic PPL1 neurons (includes MB-MP1, MB-V1 and MB-MV1 neurons), PPL2ab, in 118 PAM neurons per hemisphere (includes MB-M3 neurons), in PPM3, in 1 PPM1 neuron per hemisphere, in PPM2, PPL2c, VUM1 and VUM2 neurons. Expression of Scer\GAL4ple.PF is also detected in dopaminergic T1, SVP1, D1 and SP1 neurons.
Scer\GAL4ple.PF drives expression in a subset of the dopaminergic neurons in the adult brain labelled by ple : around 8 PAM neurons, 5 PAL neurons, 4 PPM1 neuron, 8 PPM2 neurons, 7 PPM3 neurons, 11 PPL1 neurons and 6 PPL2 neurons.
Expression observed in neurons that innervate the mushroom body vertical lobes and peduncles, the fan-shaped body and the protocerebral bridge.
Scer\GAL4ple.PF is expressed in approximately 12 PPL1, 11 PPL2, 8 PPM1/2, 7 PPM3 and 5 PAL dopaminergic neurons, but is not expressed well in the PAM cluster. PAL neurons are seen to project across the midline. Neurons of the PPM1/2 cluster are seen to project ventrally. PPL1 cluster neurons are seen to project medially and dorsally towards the mushroom body, and the PPM3 neurons are observed to project towards the central complex.
Approximately 127 Scer\GAL4ple.PF expressing cells can be identified in the adult protocerebrum. Scer\GAL4ple.PF is highly expressed in neurons that terminate in the vertical lobes of the mushroom body and in the lateral part of the mushroom body horizontal lobes. There are also a few fibers wrapped around the distal tip of the mushroom body horizontal lobes.
Scer\GAL4ple.PF drives expression in the dopaminergic cluster PAL, PPL1, PPL2ab, PPM1, 10 neurons of the PPM2 cluster, 8 neurons of the PPM3 cluster and 13 neurons of the PAM cluster.
Scer\GAL4ple.PF labelled neurons that innervated, particularly, the , midregion of the peduncle, the heel, alpha lobe and lateral region of the gamma lobe. Labelled processes ate also found surrounding the median portion of the beta and gamma lobes. Expression in larva is similar to adults, with Scer\GAL4ple.PF driving expression in neurons that innervate the the vertical lobes and heel.