Paths, elongation, and projections of ascending chick embryonic spinal commissural neurons after crossing the floor plate

Abstract

The paths of embryonic chick spinal commissural neurons originating from the lumbosacral (LS) 2 spinal segment and born around Hamburger-Hamilton stage (HH) 18 were observed by labeling the axons with an in ovo electroporation method designed to limit the electroporated area to approximately one somite length. After crossing the floor plate, these axons followed two major paths, one ventral and one dorsal, and a minor path running between the major ones. These axons reached the brachial region by HH28, passed through the cervical region at HH29, and entered the medullary area by HH30. The dorsal axons entered the developing cerebellum by HH33, crossed the midline again, and spread into the rostral-ipsilateral area of the developing cerebellum so that most of them were confined to lobules II-III by HH39. A small population of ventrally running axons turned to enter the cerebellum, and the rest entered the superior medullary velum between the cerebellum and the midbrain. The LS2-originating axons that ascended ipsilaterally into the cerebellum followed a single path, and their extension was delayed compared with that of the commissural axons. Some of the ipsilateral axons innervated the cerebellum; the rest entered the superior medullary velum. These direct observations of the formation of part of the spinocerebellar projections in chick will be a useful reference for future analyses of the underlying mechanisms.