Extended Data Fig. 6: PFL3 neurons can be distinguished from PEG neurons based on their electrophysiological properties and individual PFL3 neurons are tuned to heading, with different cells showing different preferred heading angles.

a, Biocytin fill of a PFL3 neuron (left) and a PEG neuron (right) recorded in the split-Gal4 line VT000355-AD ∩ VT037220-DBD. PEG and PFL3 neurons can be differentiated based on their innervation patterns. Specifically, PFL3 neurons innervate the fan-shaped body (FB) and lateral accessory lobe (LAL) whereas PEG neurons innervate the ellipsoid body (EB) and the gall (GA). Each image is a maximum z-projection from a subset of slices. One of eight PFL3 cells and one of three PEG cells in which such a fill was visualized is shown here; in most recordings we used the electrophysiological properties of the neuron recorded to identify it as a PFL3 or PEG cell (Methods). b, Sample Vm from the PFL3 and PEG neuron depicted in the anatomy panels directly above. At depolarized membrane potentials, the spikes of PFL3 neurons were relatively small (left) whereas those from the PEG neurons were relatively large (right). Black dots indicate detected spikes. c, At hyperpolarized membrane potentials, PFL3 neurons display rhythmic oscillations (left), whereas the membrane potential of PEG neurons tends to be more flat (right). d-e, Vm (spikes removed) (left) and spike rate (right) tuning curves to heading direction for all PFL3 cells. Dashed line in the Vm curves represents a sinusoidal fit to data, which was used for estimating the cell’s preferred-heading direction (see Methods). Shaded area represents 90° gap at the back of the arena where the bar is not visible. Cells are sorted and numbered based on their estimated preferred-heading direction. We use this numbering scheme throughout the manuscript to refer to specific cells.

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