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Fig. 8. Early-born neurons and Sema3a repel TrkA⁺ afferents in the dorsal spinal cord. (A,B) TrkA immunocytochemical staining of E14.5 wild-type (A) and Dcc^–/– mutant (B) spinal cords shows an absence of TrkA ⁺ afferents in the medial superficial dorsal horn of Dcc^–/– mutant (arrow in B) when compared with wild-type control (arrow in A). Brackets in A-D indicate superficial dorsal horn. (C,D) In situ hybridization in wild-type (C) and Dcc^–/– mutant (D) dorsal horn shows the ectopic Sema3a in the medial superficial dorsal horn of the mutant (arrow in D) when compared with wild-type control (arrow in C). (E) Immunocytochemical staining of TrkA⁺ afferents (red) and EGFP (green; arrow in E) in Sema3a+EGFP co-electroporated spinal cord. (F) Strong Sema3a expression in electroporated spinal dorsal horn. Arrow indicates introduced Sema3a expression, whereas arrowhead indicates endogenous Sema3a expression in the ventral horn. (G,H) Higher magnifications of E, showing TrkA staining of the contralateral (G) and electroporated sides (H) of the spinal cord. On the contralateral side of the dorsal horn, strong TrkA staining is detected in laminae I-II (arrowheads in G), but on the electroporated side much less TrkA staining is seen in laminae I-II (arrowheads in H). (I) Quantitative analysis of TrkA-positive areas. Asterisk indicates a significant difference in the area of TrkA-positive staining. ^*P<0.001. Areas of TrkA⁺ fibers are expressed by k µm²; k=1000. Co, contralateral side; Ep, electroporated side. Scale bars: 200 µm in E,F; 100 µm in A-D,G,H.

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