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. Author manuscript; available in PMC: 2014 Feb 1.
Published in final edited form as: Neurogastroenterol Motil. 2012 Dec 18;25(2):e89–e100. doi: 10.1111/nmo.12057

Figure 4.

Figure 4

GFP-ir fibers in a nerve strand and a globular cluster within the myenteric plexus. Images show triple labeling for GFP (green), CGRP (red), and SP (blue) with DAPI nuclear counterstain (gray) and are projections of 17 (A–D), 16 (E–H), or 12 (I–L) slices 1 μm apart. A–D, In a nearly horizontal section adjacent to the section displayed in Fig. 3, GFP-, CGRP-, and SP-positive fibers are seen in a nerve strand overlaying the longitudinal muscle (LM). Circular muscle (CM) and portions of myenteric ganglia (MG; arrow in C) are seen on each side of LM. E–H, A portion of the nerve strand in A–D shown at a higher magnification. Insets show a single slice from a region at the end of the nerve strand. In E, a single large axonal profile is indicated by small arrows. Colocalization of GFP-ir with CGRP- and SP-it is seen in some small –diameter axons (arrows in F and G). In D and H, the GFP-, CGRP-, and SP-positive fibers are shown relative to DAPI nuclear staining. Note the elongated nuclei of smooth muscle cells in the LM and CM oriented perpendicular to each other. Spherical nuclei are seen in the MG. I–L, A transverse section showing a globular cluster of GFP- and CGRP-ir fibers (I and J) within a MG that is demarcated by SP-ir (K). A single slice is shown in the insets. Two GFP-positive fibers appear to contribute to the cluster (I). The smaller of these fibers also shows CGRP-ir (see arrow in J). A single slice shown in the insets in J demonstrates that the cluster contains fibers that are double-labeled for GFP and CGRP as well as single-labeled GFP- and CGRP-ir fibers. The contribution of SP-ir fibers to the cluster is minimal (insets in K). In L, the GFP-ir cluster in shown relative to the nuclei of the LM and CM. Note that in the tranverse section nuclei of CM cells appear elongated, whereas LM nuclei are seen in cross-section. Nuclei of the MG are seen between the layers of LM and CM nuclei. A single slice in the inset shows a nucleus in the center of the clustered fibers. Scale bars: A–D, 50 μm; E–L, 10 μm.