Table 1 Morphological features of photo-labeled projection neurons across species

	Neuron number			Bouton cluster volume (μm3)
	D. melanogaster	D. simulans	D. sechellia	D. melanogaster	D. simulans	D. sechellia
DA2	4 ± 0.80 lPNs 0 vPN	4 ± 1.02 lPNs 0 vPN	4 ± 0.40 lPNs 0 vPN	112.79 ± 11.30	110.95 ± 37.06	179.18 ± 31.76
DC3	3 ± 0.29 adPNs 0 vPN	3 adPNs 3 vPNs	2 ± 0.29 adPNs 0 vPN	370.15 ± 195.68	364.06 ± 146.60	233.02 ± 83.80
DL2d	5 adPNs 1 vPN	5 adPNs 1 vPN	5 adPNs 3 vPNs	112.18 ± 12.41	104.44 ± 25.93	348.29 ± 186.63
DM2	2 lPNs 0 vPN	2 lPNs 0 vPN	2 lPNs 0 vPN	367.38 ± 54.87	309.52 ± 89.42	365.40 ± 52.39
DP1l	1 lPN 1 vPN	1 lPN 2 vPNs	1 lPN 2 vPNs	244.39 ± 69.00	232.41 ± 100.73	388.91 ± 75.88
DP1m	1 adPN 0 vPNs	1 adPN 0 vPNs	1 adPN 0 vPNs	221.59 ± 72.28	172.85 ± 113.80	260.95 ± 105.19
VA1d	3 adPNs 1 vPN	3 adPNs 1 vPN	3 ± 0.40 adPNs 0 vPN	205.89 ± 26.65	195.78 ± 109.79	155.87 ± 50.25
VC3	3 adPNs 0 vPN	3 adPNs 0 vPN	3 adPNs 0 vPN	200.70 ± 97.03	158.48 ± 90.40	143.59 ± 46.17
VL2a	1 adPN 3 vPNs	1 adPN 3 vPNs	1 adPN 3 vPNs	174.36 ± 64.22	156.05 ± 79.93	208.92 ± 60.55
VM5v	3 ± 0.49 adPNs 0 vPN	3 ± 0.40 adPNs 0 vPN	2 ± 0.33 adPNs 0 vPN	256.78 ± 104.09	268.43 ± 132.67	140.14 ± 77.76

Morphological features of projection neurons—namely the number of neurons associated with a given glomerulus and the volume of the presynaptic sites or boutons these neurons form in the mushroom body—were measured and compared across species (n = 5 for each type of projection neuron, standard deviation from mean is shown). Projection neurons showing significant shifts in connectivity frequencies (DC3, DL2d, VM5v, DP1l, VA1d, VC3, and VL2a projection neurons) were analyzed as well as some projection neurons that did not show significant shifts in connectivity frequencies (DA2, DM2, and DP1m projection neurons). Projection neurons were typed based on whether their cell bodies are located in the anterior-dorsal (ad), lateral (l) or ventral cluster (v). All source data used in this table are provided in the Source Data file.

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