Time-dependent behavior of neurons, resident microglia, and astrocytes in injured spinal cords. Cross sections (30 μm) were obtained from the intact spinal cord (A), epicenter (Epi, B), P1 (C), and P2 (D) areas in the caudal direction at the indicated times after injury. The behaviors of neurons (white arrowheads), astrocytes (white arrows), and resident microglia/monocytes (black arrows) were compared by staining immediately adjacent serial sections with antibodies against NeuN, GFAP, and Iba-1, respectively, and then visualizing using HRP-conjugated secondary antibodies. Dorsal (D), medial (M), and ventral (V) areas were magnified in the lower panels. Data shown are representative of more than three independent experiments. In each experiment, at least five animals were used for each time point. Scale bar: 200 μm (upper panels), 50 μm (lower panels).