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Fig. 2. Kidins220 localization pattern in somites and spinal neurons of DAB-stained embryos. Lateral views, anterior to the left, dorsal to the top (A-D). Sagittal section of a Kidins220-immunostained embryo (A) and Hoechst-counterstaining (B). The red square brackets indicate paraxial mesoderm. Whole-mount ab34790 anti-Kidins220-immunostaining (C) and Ab12101 anti muscle-immunostaining (D). Coronal sections of Kidins220 (Hoechst-counterstained) and Ab12101-stained embryos, respectively (E,F). The arrows in (E) show the Kidins220 signal in lateral regions of spinal cord. High magnification of a Kidins220-positive intersomitic junction (G). Transverse sections of Kidins220 and acetylated tubulin-immunostained embryos, respectively (H,I): the arrowheads show the exit point of a motor neuron that in (H) resulted broken because of sectioning. Sagittal section of a stage 42 embryo (J) and Hoechst-counterstaining (K). The arrowheads in (J) indicate motor neurons. The square brackets in (E,J,K) span somite length. Acet. tub.= acetylated tubulin; nt= neural tube; ij= intersomitic junction. Scale bars: (A,B,H,I) 50 um; (C,D) 250 um; (E,F) 25 um; (G,J,K) 10 um. |
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Fig. 5. Sagittal cryosections of stage 42 embryos double-labeled for Kidins220 (red) and acetylated tubulin (green) by immunofluorescent staining. Kidins220-positive signal was present in dorsal and ventral regions of the spinal cord as well as in motor neurons (arrows) (A) as shown by the acetylated tubulin staining (B,C). Magnification of motor neurons double-stained by Kidins220 (D), acetylated tubulin (E) and merged (F). The dashed bar in (D) spans the somite length. Kidins220 protein in eye of st. 33-34 DAB-stained embryos (G) and st. 42 embryos double-immunolabeled for Kidins220 and acetylated tubulin (H-J). The open arrowhead in (H,J) indicates the Kidins220 signal in the optic nerve. The dashed circle indicates the position of lens in the eye. The white arrow in (H,J) denotes the presence of Kidins220 signal in a facial muscle near the eye. The white arrowheads in (H,J) indicate Kidins220 and acetylated tubulin-positive nerve fibers present in epidermis. ep, epidermis; l, lens; rgc, retinal ganglion cell; s, somite; sc, spinal cord; *, ciliated cell. Scale bars: (A-C) 50 um; (D-F,G-J) 25 um. |
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Fig. 4. Whole mount immunohistochemistry experiments on stage 42 embryos. Lateral view of a posterior portion of DAB-stained embryo for Kidins220 (A). Parasagittal section performed laterally between the skin and the somites of an immunostained embryo (B) and Hoechst-counterstaining (C). Dashed lines in (A,B) highlight the pattern of the Kidins220 signal, organized in irregularly parallel rows to the side of intersomitic junctions. The square brackets in (B,C) span somite length. Lateral view of somite region of whole-mount double immunofluorescent staining for Kidins220 (red signal) and acetylated tubulin (green signal) (D-F). Magnified views of neurons in somite region (G-I), dorsal fin (J-L; M-O), abdominal region (P-R). The largest dashed box in (D) indicates the region magnified in (G-I); the smallest one, the region magnified in (J-L). White arrowheads in (G,I,J,L,M,O) highlight the spotted signal of kidins220 at some neural fiber. White arrowheads in (M,O) show specific cells in dorsal fin, positive for both Kidins220 and GFAP. The asterisks in (E,H,K) denote the acetylated tubulin-positive but Kidins220-negative ciliated cells. Scale bars: (A) 125 um; (B-F,J-O) 50 um; (G-I,P-R) 10 um. |
