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. 2013 Sep 1;140(17):3595–3600. doi: 10.1242/dev.091033

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© 2013. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 3. — Neural crest corridor development. (A-C) Lateral view of Sox10/Isl1 double-labelled chick embryos showing NCC corridor development in relation to developing CSG. (A′-C′) Transverse sections reveal extension of Sox10⁺ NCCs (yellow arrowheads indicate ventral point) to placode (asterisks) with increasing neurogenesis. (D) Transverse section of triple-labelled HH17 chick (D) shows HNK1⁺ NCCs (magenta; white arrowhead) and Foxd3⁺ NCCs (yellow; yellow arrowhead), surrounding ISL⁺ neuroblasts (green) (confocal stack projection). (E-G) Coronal sections show restriction of HNK1 staining to periphery of NCC corridor (confocal stack projection). At HH13, NCCs are Sox10/HNK1 double positive (E), but by HH14⁺ (F) HNK1 staining (magenta; white arrowhead) is localised to the periphery of Sox10⁺ NCCs (yellow; yellow arrowhead). At HH17-, ISL1⁺ neuroblasts can be seen at core of NCC corridor (G). (H) Coronal section of GFP NCC electroporated chick confirms restriction of HNK1 staining (magenta; white arrowhead) to periphery of total GFP⁺ NCCs (green; blue arrowhead) at HH14 prior to robust neurogenesis (single optical confocal section). (I) Analysis in mouse at 18 ss also shows YFP⁺ NCCs (magenta; orange arrow) forming corridors containing few NFM⁺ processes (green; white arrow). (J) Schematic representation of the NCC corridor with HNK1⁺ NCCs (magenta) surrounding Sox10⁺ NCCs (yellow), extending to the placode (asterisk). Neuroblasts (blue) migrate at the NCC corridor centre. Arrow indicates direction of migration. (K,K′) Chick hindbrain explants demonstrate in vitro streamed NCC migration with DIC (K) and HNK1 (K′). (L,L′) Confocal analysis reveals 3D nature of in vitro HNK1⁺ NCC corridor (L), whereas digital re-section reveals peripheral HNK1 staining (L′). Asterisks indicate placode; NT, neural tube; OV, otic vesicle; PAI, PAII, pharyngeal arches I and II. Scale bar: 100 μm.