(A) Experimental preparation. We targeted single neurons for patching using cell type-specific expression of GFP. Flies were placed in an arena equipped with rotatable stimulus delivery and live imaging of behavior. All data shown are from awake non-flying animals. (B) Stimulus details. Left: cue presentation directions. Front (0°, gold), rear (180°, brown), ipsilateral (90°, black), and contralateral (−90°, gray) to the recorded neuron. Right: stimulus validation. Each plot shows measurements from a photodiode (top), anemometer (middle), and photo-ionization detector (PID, bottom). PID units are arbitrary. The five stimulus combinations were: a high contrast stripe illuminated by 15 mW/cm2 ambient lighting (red), a 25 cm/s airflow stream (blue), stripe and airflow together (purple), airflow and 20% apple cider vinegar together (orange), and all three modalities simultaneously (green). Each trace is 12 s long. Simultaneous cues were presented from the same direction. (C) Rhythmic and tonic baseline activity in a subset of CX columnar neuron types. Left: raw membrane potential over time for three example neurons. P-EN2 and P-F2N3 show rhythmic activity at different frequencies, while P-F3LC fires tonically at rest. Right: resting membrane potential probability distributions for each recorded neuron of the types shown (gray). Example neurons in black. Rhythmic neurons exhibit broad distributions, while tonic neurons show tight distributions. See also Figure 1—figure supplement 1. (D) Left: CX neuropils innervated by P-F1N3 neurons (gray). PB, protocerebral bridge; FB, fan-shaped body; EB, ellipsoid body; NO, noduli. Right: PSTHs for a single P-F1N3 neuron. Each trace represents the mean of four presentations of stripe alone (red, top) or airflow alone (blue, bottom) from one direction. Colors representing different directions as illustrated in (B). Colored boxes indicate the 4 s stimulus period. Dashed line indicates 0 Hz. (E) Responses to airflow (blue) versus stripe (red) for each neuron type. Gray dots indicate the mean spiking response of each cell (1 s stimulus minus 1 s baseline) to four trials from the direction producing the strongest response (see Materials and methods). Colored bars: mean across cells. The example P-F1N3 neuron from (D) is shown in black. Significant differences (p<0.05 by sign-rank test) between modalities are marked with an asterisk. For additional detail, see Fig. S2. (F) Left: CX neuropils innervated by P-F3N2d neurons. Right: PSTHs for a single P-F3N2d neuron. Each trace represents the mean of four presentations of airflow alone (blue, top) or airflow and odor together (orange, bottom) from one direction. Plot details as in (D). (G) Responses to odorized airflow (orange) versus airflow alone (blue) for all cell types recorded. Asterisk: odor significantly reduces the response of P-F1N3. Plot details as in (E). For additional detail, see Figure 1—figure supplement 2. (H) Mean airflow response across cells as a function of airflow direction for each cell type. Cell types are plotted in groups of two (gray, black) according to broad anatomical similarities. Note different vertical axis scales. Data at −180° is replotted from 180° for clarity (orange stars). For additional detail, see Figure 1—figure supplement 2.