Following complete transection of the thoracic spinal cord at various times during embryonic development, chick embryos and posthatched animals exhibited various degrees of anatomical and functional recovery depending upon the age of injury. Transection on embryonic day 2 (E2), when neurogenesis is still occurring and before descending or ascending fiber tracts have formed, produced no noticeable behavioral or anatomical deficits. Embryos hatched on their own and were behaviorally indistinguishable from control hatchlings. Similar results were found following transection on E5, an age when neurogenesis is complete and when ascending and descending fiber tracts have begun to project through the thoracic region. Within 48 h following injury on E5, large numbers of nerve fibers were observed growing across the site of transection. By E8, injections of horse-radish peroxidase (HRP) administered caudal to the lesion, retrogradely labelled rostral spinal and brainstem neurons. Embryos transected on E5 were able to hatch and could stand and locomote posthatching in a manner that was indistinguishable from controls. Following spinal cord transections on E10, anatomical recovery of the spinal cord at the site of injury was not quite as complete as after E5 transection. Nonetheless, anatomical continuity was restored at the site of injury, axons projected across this region, and rostral spinal and brainstem neurons could be retrogradely labelled following HRP injections administered caudal to the lesion. At least part of this anatomical recovery may be mediated by the regeneration or regrowth of lesioned axons. Although none of the embryos transected on E10 that survived to hatching were able to hatch on their own, because several sham-operated embryos were also unable to hatch, we do not attribute this deficit to the spinal transection. When E10-transected embryos were aided in escaping from the shell, they were able to support their own weight, could stand, and locomote, and were generally comparable, behaviorally, to control hatchlings. Repair of the spinal cord following transection on E15 was considerably less complete compared to embryos transected on E2, E5, or E10. However, in some cases, a degree of anatomical continuity was eventually restored and a few spinal neurons rostral to the lesion could be retrogradely labelled with HRP. By contrast, labelled brainstem neurons were never observed following E15 transection. E15 transected embryos were never able to hatch on their own, and when aided in escaping from the shell, the hatchlings were never able to stand, support their own weight or locomote.(ABSTRACT TRUNCATED AT 400 WORDS)

The ontogeny of intersegmental (propriospinal) projections was studied in the chick embryo spinal cord between embryonic day 2.5 and day 6. Our goals were 1) to determine the earliest projections of intersegmental interneurons between specific spinal regions and to establish the cell types involved; and 2) to follow the ontogeny of these projections during the early formative stages of spinal cord development. Studies were carried out in vitro by using an isolated spinal cord/brainstem preparation. Horseradish peroxidase injections were made either uni- or bilaterally at various levels of the spinal cord along the rostrocaudal axis of the embryo. HRP histochemistry was done on Vibratome sections with diaminobenzidine as the chromogen. Following unilateral injections at day 2.5, labelled commissural interneurons were found contralaterally and were confined to the injected segment. Subsequently, labelled cells were found progressively further away from the injected segment. By day 4.5 reciprocal projections extended between lumbar and brachial regions. Interneurons with intersegmental axonal projections were often undifferentiated, consisting of primitive unipolar or bipolar cells with little, if any, dendritic development. In some cases migrating interneurons could be retrogradely labelled from two or three segments away from the location of their translocating cell body. Anterograde Golgi-like labelling of early undifferentiated cells revealed growing axons, axonal terminals, and growth cones. Five or six reasonably distinct classes of intersegmental interneurons were identified based on their location, axonal projections, and morphology of dendritic arbors. These appeared to be segmentally and bilaterally arranged along the rostrocaudal axis of the spinal cord. The axons of some of these types of interneurons exhibited preferences in their longitudinal projections within the ventral and ventrolateral marginal zone at the very onset of pathway formation. From the present observations it can be concluded that intersegmental connectivity precedes the development of ascending and descending supraspinal, as well as primary afferent connections in the chick embryo spinal cord.

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