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. 2013 Sep 18;8(9):e74454. doi: 10.1371/journal.pone.0074454

Figure 7. Spinal labeling of last-order (premotor) interneurons with PRV, co-labeling with ChAT, and contacts with CSTs.

Figure 7

(A) Example of labeling in one 40 µm section achieved at 64 hours after intramuscular PRV injection. Large panel shows labeling on ipsilateral side (Calibration: 500 µm). There was minimal contralateral labeling. Top inset shows an example of labeled interneurons (located in lamina 4 on the section shown), and lower panel, a motoneuron, at higher magnification (Calibration: 25 µm). (B1–B2) Overlaid section images for two representative animals processed in Neurolucida showing positions of individual last-order interneurons from PRV injected into the deltoids, biceps and wrist extensors at levels C5/C6 (B1) and C7/C8 (B2) that also label positively for ChAT. (C1–C2) Confocal images of two representative PRV-ChAT double-labeled interneurons at levels C7/C8 (C1) and C5/C6 (C2). ChAT = red; PRV = green. (D1–3) (D) Confocal images of PRV-labeled interneurons receiving contacts from BDA-labeled CST axons terminals. PRV was injected into the biceps and wrist extensor compartments. Each panel shows a projection image (center, large image) and representative 1 µm optical slices (insets). Arrows show sites of contact,. Scale bars for A, large panel = 500 µm, smaller panels = 50 µm; B, same as A; C and D = 20 µm.