Extended Data Fig. 11: Spilt-GAL4 lines targeting oviDN input neurons and analysis of oviDN inputs in the hemibrain connectome.

a-r, Average z-projection of oviDN input split-GAL4 lines in the brain (top) and ventral nerve cord (bottom) in order of x-axis in Fig. 6a (see Supplementary Table 3 for additional information). Green shows UAS-CsChrimson-mVenus expression in the targeted neurons and magenta represents a neuropil counterstain (Methods). s, In panels s-v, we show circuit motifs that are not supported by our analysis of the hemibrain connectome¹⁶, in contrast to the motif reported in Fig. 6e, which is supported (see Methods for more discussion). This panel shows that no chemical-synapse-based recurrent circuit is observed between the oviDNs themselves in the hemibrain connectome at a threshold of ≥ 10 synapses per connection (even true at a threshold of ≥ 2 synapses here). (For reference on the potential functional significance of a 10 synapse threshold, ~15-20 neurons make ≥ 10 synapses onto an individual oviDN in the hemibrain.) Scatter plot of connections between pairs of neurons is shown to right. Both data points are from a single pair of oviDNs. t, We found in the hemibrain connectome all pairs of cells that fulfilled the circuit diagram shown on the left at a threshold of ≥ 1 synapse per connection. No direct, bidirectional, chemical-synapse-based recurrent circuit could be detected between individual oviDNs and any oviDN input neuron in the hemibrain connectome at a threshold of ≥ 10 synapses per connection (even true at a threshold of ≥ 4 synapses per connection here). Scatter plot of connections shown to right; each dot represents the connections between two neurons. Orange points represent pairs of connected neurons diagrammed in the recurrent circuit in Fig. 6e, but assayed for participation in a different circuit motif here. u, We found in the hemibrain connectome all sets of four cells that fulfilled the circuit diagram shown on the left at a threshold of ≥ 1 synapse per connection. None of these putative circuits had ≥ 10 synapses for all four connections, which we interpret to mean that no chemical-synapse-based, disynaptic recurrent circuit exists between individual oviDNs and a single class of oviDN input neuron. Cell classes (types) were based on the hemibrain v1.2.1 connectome annotation¹⁶. Scatter plot of connections is shown to right; each dot represents the connections between a set of four neurons. Orange points represent sets of four neurons diagrammed in the recurrent circuit in Fig. 6e, but assayed for participation in a different circuit motif here. For example, the orange dot indicated by an arrow represents the following connections between single cells: oviDNa(right)←groupU(right)↔groupU(left)→oviDNb(left). v, Same as panel u, but for a different circuit architecture. Each point, once again, represents the connections between a set of four neurons. None of these putative circuits had ≥ 10 synapses for all four connections.