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First published online 17 November 2004
doi: 10.1242/dev.01533

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Neural crest cell lineage segregation in the mouse neural tube

¹ Department of Anatomy and Cell Biology, University of Melbourne, Melbourne, Victoria 3010, Australia
² UMR 955 INRA de Genetique Moleculaire et Cellulaire, Ecole Nationale Veterinaire d'Alfort, Maisons-Alfort Cedex, France

View larger version (44K): [in a new window]   Fig. 1. Development of ßgal expression in dorsal regions of WlacZ/+ embryos. (A) Low-power view through the trunk section of an E9.5 embryo. (B) High-power view reveals faint ßgal+ cells (arrow) within the dorsal midline at E9.5. (C) Low-power view through the trunk at the level of the forelimb of an E10.5 embryo. (D) High-power view of the dorsal midline reveals strong ßgal+ cells (filled arrow) on the dorsal midline of the NT. (E) High-power view of the ectoderm reveals strong ßgal+ cells (filled arrows). There are also ßgal+ cells in ventrolateral regions of the NT and surrounding it (open arrows in C and E). ect, ectoderm. Scale bars: 250 µm in A,C; 50 µm in B,D,E.

View larger version (111K): [in a new window]   Fig. 2. Histochemical staining for ßgal+ in wholemounts of WlacZ/+ mice. (A) At E9.5, ßgal+ cells (arrows) are present in the cervical region of the embryo on the dorsal midline; no ßgal+ cells were observed elsewhere in the ectoderm. (B) Low-power view of an E10.5 embryo showing strong ßgal+ cells along the dorsal midline (arrows), and lateral to it under the ectoderm. (C) High-power view of the boxed area in B, showing ßgal+ cells on the dorsal midline and emanating from it (arrows). (D) Montage of images from the trunk of an E11 embryo, showing ßgal+ cells distributed extensively under the ectoderm. dm, dorsal midline. Scale bars: 0.5 mm.

View larger version (123K): [in a new window]   Fig. 3. Kit+ cells that migrate dorsolaterally are melanocyte progenitors. (A) Low-power view of the dorsal region of an E11 WlacZ/+ embryo stained for ßgal (red) and Trp2 (green). (B) High-power view of the region outlined in A. ect, ectoderm. Scale bar in B: 50 µm.

View larger version (90K): [in a new window]   Fig. 4. Kit+ cells in and around developing sensory ganglia co-localise with blood vessels. (A) Section through a dorsal root ganglion in an E10.5 WlacZ/+ embryo, stained for CD34. (B) Section of an E10.5 WlacZ/+ embryo stained for F4/80 in an area just adjacent to ganglion. (C) Same region as is shown in B, stained for ßgal. (D) Merged views of B and C. (E) View of the dorsolateral region of an E10.5 WlacZ/+ embryo stained for CD34 + F4/80 (red), revealing the blood vessel network, and ßgal (green). Bright ßgal+ cells are seen under the ectoderm (closed arrow), as well as pale ßgal+ cells associated with blood vessels (double-headed arrow). Some ßgal+ cells are also present within the NT. Dotted lines indicate either the surface of the embryo, the location of the ganglion, or the border of the NT. bv, blood vessel; drg, dorsal root ganglion. Scale bars: 50 µm in A-D; 150 µm in E.

View larger version (108K): [in a new window]   Fig. 5. p75 and ßgal staining reveal separate populations of NC precursor cells in the trunk NT. WlacZ/+ embryos were stained for p75 using immunohistochemistry and ßgal histochemistry, from stages E9 to E11. Low-power views of these embryos are shown at the left of the panel for each age. Extent of ßgal expression on the dorsal midline of the NT is indicated by dotted lines. High-power dorsal views of cervical, midtrunk and lowertrunk regions of these embryos are shown in the middle of each age panel. For these views, a negative of the bright field image is shown with ßgal staining false-coloured red; each image has been merged with the p75 fluorescence image (green) of the same area. Coronal sections were taken through cervical, midtrunk and lower trunk regions (approximate locations are indicated by bars in low-power views of embryos) and double stained by fluorescence immunohistochemistry for p75 (green) and ßgal (red). These are shown at the right of each age panel. High-power views of the dorsal midline of the NT from these sections are inset into the images where there is any indication of both ßgal and p75 staining in this region. Scale bar: 100 µm.

View larger version (76K): [in a new window]   Fig. 6. Analysis of WlacZ/+ embryos in the head shows that Kit+ cells at the dorsal midline of the midbrain-hindbrain junction give rise to melanocytes. (A) Dorsolateral view of the rostral region of a whole-mount embryo at E10.5, showing ßgal+ cells emanating from the dorsal aspect of the midbrain-hindbrain junction. ßgal+ cells emanate caudolaterally as a stream through the mesenchyme adjacent to the hindbrain, as well as to the ectoderm (asterisk). (B) Transverse section through an E10.5 embryo at the midbrain-hindbrain junction, showing ßgal+ cells on the dorsal midline and ectoderm. A high-power view of the same region stained for p75 and ßgal is shown in the inset; strong p75 staining is on surface of embryo. (C) Lateral and (D) dorsal views of an E11 embryo, showing increasing distribution of ßgal+ cells through ectoderm (asterisk). (E) Transverse section through the rostral hindbrain of an E10.5 embryo at level indicated by broken line in C, stained for ßgal (red) and Trp2 (green). The border between the mesenchyme and neural tube is indicated with a dotted line. Some ßgal+ cells are also present on base of roof plate (A,C, open arrow). rp, roof plate; st, stream. Scale bars: 500 µm in A,C,D; 200 µm in B; 50 µm in E.

View larger version (75K): [in a new window]   Fig. 7. Non-melanogenic Kit+ cells in the head are not derived from Kit+ dorsal midline NT cells. (A) Dorsolateral view of a whole-mount E9.5 WlacZ/+ embryo. ßgal+ cells are present in the branchial arches and presumptive cranial ganglia (filled arrows), and on roof plate (open arrow), but no ßgal+ cells emanate from the midbrain-hindbrain junction. (B) Transverse section through an E9.25 embryo at the midbrain-hindbrain junction, showing absence of ßgal+ cells on the dorsal midline and ectoderm. (C) Low-power view of a transverse section through the hindbrain region of an E10 WlacZ/+ embryo at level of the trigeminal ganglion (boxed) stained for ßgal (green) and Isl1 (red). (D) High-power view of the boxed area in C, showing that many cells in the trigeminal ganglion are ßgal+/Isl1+ (yellow). (E) Transverse section through the caudal region of an E10.5 WlacZ/+ embryo. Strong ßgal+ cells are present on roof plate, but completely absent in the dorsal neuroepithelium. Scale bars: 200 µm in A-C,E; 50 µm in D.

View larger version (49K): [in a new window]   Fig. 8. Scheme for the development of Kit+ and p75+ cells in the developing premigratory and peripheral NC. Cells inside the NT represent cells in the premigratory crest, other cells represent migrated NC cells, and those on the borders of embryo represent NC cells in the ectoderm.