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. 2016 Feb 15;5:e13253. doi: 10.7554/eLife.13253

Figure 4. Circuit diagram around GDLs.

(A) Comparing the confocal images (left) and EM reconstruction (right) of a GDL (top: cross-sectional view, bottom: dorsal view). Postsynaptic sites (cyan) and presynaptic sites (red) are shown in the EM images (right). Scale bar represents 20 μm (upper left), 10 μm (bottom). (B) The EM circuit graph of GDLs and their postsynaptic neurons. Hexagonal shape denotes a group of left-right homolog neurons. Connections with less than 6 synapses are not included (green: premotor neurons, yellow: others). (C) Major postsynaptic (“downstream”) targets of a GDL in the abdominal segment A1. A27h is the strongest synaptic partner of a GDL. Numbers on the directed arrows indicate the number of synapse. (D) Major presynaptic (“upstream”) targets of a GDL include two dendritic arborization (da) sensory neurons (blue). All other presynaptic targets identified are premotor neurons in the posterior segments (green). (E) A circuit model around GDLs. From the wiring diagram, a GDL has connections with several premotor neurons at both upstream and downstream. The symbols: NMJ (arrowheads), putative excitatory synapse (circles), and inhibitory synapse (bars). Thickness corresponds synaptic strength. (See also Figure 4—figure supplement 1–3.)

DOI: http://dx.doi.org/10.7554/eLife.13253.011

Figure 4.

Figure 4—figure supplement 1. No GRASP signal was detected with motor neurons (Related to Figure 4).

Figure 4—figure supplement 1.

(A–D) GRASP experiments. (A, B) Expression pattern of the driver lines 9-20-GAL4 and OK6-LexA were assessed with 10xUAS-IVS-mCD8::RFP,13xLexAop2-mCD8::GFP and immunostaining with anti-GFP (green), anti-DsRed (yellow), anti-FasII (magenta) antibodies. Arrows denote GDLs in (B). (C) Results of GRASP (9-20-GAL4,OK6-LexA>OK6-LexA, LexAop-CD4::spGFP11;UAS-CD4::spGFP1-10) visualized by immunolabeling with a monoclonal antibody against GFP (Sigma). No GRASP signal was detected. Dashed box indicates the positions of the presynaptic sites of GDLs. (D) Syt::HA was coexpressed with GRASP reporters to assess the precise location of GDL presynaptic terminals (UAS-syt::HA;9-20-GAL4,OK6-LexA>OK6-LexA,LexAop-CD4::spGFP11;UAS-CD4::spGFP1-10). However, no GRASP signals were observed at the terminals (arrows). Scale bar represents 30 μm in (A–C) and 20 μm in (D).
DOI: http://dx.doi.org/10.7554/eLife.13253.012
Figure 4—figure supplement 2. Morphology of the major presynaptic and postsynaptic neurons of GDL.

Figure 4—figure supplement 2.

(A) An aCC motor neuron in A1 segment is shown in the comprehensive EM dataset. We use the position of the aCC for reference in (B) and (C). (B) Seven major presynaptic targets of a GDL (A1: vdaA, vdaC, A2: A02j, A03a4, A27h, A27k, A3: A02j). (C) Seven major postsynaptic targets of a GDL (A02d, A05k, A08e3, A27a, A27h, A31d, A31x: all of the neurons exist at A1 segment).
DOI: http://dx.doi.org/10.7554/eLife.13253.013
Figure 4—figure supplement 3. Adjacency matrix for GDL circuits.

Figure 4—figure supplement 3.

Each row and column represents a neuron's’ pre- and post- synaptic contacts, respectively. The number in the matrix is the synapse number between the target neurons and their partners. Neurons are grouped by segments. GDLs are annotated as “A27j2_a1” in this figure.
DOI: http://dx.doi.org/10.7554/eLife.13253.014