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Fig. 3 Spiral fiber neurons. (A1) Pairwise recording of a GFP-negative spiral fiber neuron at rhombomere 3 and the contralateral M-cell in the Tg(glyt2:GFP) x relaxed line at 4 dpf (Left). (A2) Firing the spiral fiber led to EPSPs (black) in the M-cell. The slow component of the EPSPs was blocked by the glutamatergic blockers 10 μM NBQX and 100 μM d-AP5 (gray). (A3) PSP in the spiral fiber neuron in response to firing the M-cell. (A4 and A5) Top-down and cross-section views of the spiral fiber neuron (green) and the M-cell (red) from A2 and A3 relative to the glycinergic neurons (blue). Arrowheads mark the glycinergic stripes. (B1 and B2) Summary of the locations of eight spiral fiber neurons (blue) in the glycinergic line in rhombomeres 4 and 5, laid out as in Fig. 2B. (C1) Cross-section view of the backfilled spiral fiber neurons (red) in the transgenic glutamatergic line [Tg(vglut2.1:GFP)] at rhombomere 3. Arrowheads mark the glutamatergic stripes. (C2) Cross-section of the backfilled spiral fiber neurons (white) in the Tg(vglut2.1:RG x dbx1b:Cre) line. Arrowhead indicates the dbx1b-positive middle glutamatergic stripe (green). (D1 and D2) Projections of the spiral fiber neurons shown as in Fig. 2C. Asterisk on D1 marks a point on the line for the neuron in D2. (Scale bars, 20 μm.)