Development of the major pathways for neurite outgrowth in the chick hindlimb

To elucidate mechanisms that may control development of the gross anatomical nerve pattern, motoneuron outgrowth into the chick hindlimb was examined using orthograde labeling, scanning and transmission electron microscopy, and Alcian blue staining. Results show that growth cones are not guided by contact with oriented extracellular fibrils, aligned mesenchyme cells, the myotome, or the vasculature. Pathways are not delineated by cell-free space or channels of lower cell density; however, densely packed mesenchyme may form barriers that channel outgrowth. In addition, abundant mesenchymal cell death was seen at the nerve front. This cell death may provide space that encourages growth cone advancement. Pathways often lie along interfaces between areas that stain darkly and lightly with Alcian blue, which specifically stains glycosaminoglycans, and growth cones never penetrate areas that stain intensely, such as the pelvic girdle, which is known to be a barrier to outgrowth. Leading growth cones form specialized contacts with mesenchyme cells, but the predominant contacts are interneuronal. It is proposed that the anatomical pattern of outgrowth is determined by the distribution of preferred substrata, the most preferred substratum being other neurites. Further, neurites tend to prefer loose mesenchyme to dense mesenchyme or areas rich in glycosaminoglycans.