Initial axial level-dependent differences in size of avian dorsal root ganglia are imposed by the sclerotome

We have recently shown that there is very early variation in dorsal root ganglia (DRG) size based on their axial position. From the time of their condensation at Stage (St.) 20 (Embryonic Day 3), before the onset of apoptosis, the ganglia in brachial segments 14 and 15 are more than 80% larger on the average than those in cervical 5 and 6. This difference in volume is due to increased numbers of cells in the brachial DRG. In addition, the rostrocaudal length was found to be significantly greater for brachial ganglia, and the greater length of the brachial ganglia was found to be correlated with a greater length of brachial than cervical sclerotomes. It was therefore proposed that the difference in DRG size at the time of gangliogenesis is likely to arise from colonization by a larger initial number of neural crest cells of the longer adjacent rostral sclerotomes in brachial somites. In the present work, we have performed two types of experiments to test this hypothesis. First, we have performed heterotopic grafts of segmental plate mesoderm from cervical to brachial levels and vice versa. In all of these grafts, the sclerotomes developed with a rostrocaudal extent (length) corresponding to their level of origin in the donor embryo. DRG that formed in the grafted mesoderm attained a length appropriate to that of ganglia developing in segments of the axial level of the donor mesoderm, not that of the host. Second, we have estimated proliferation of DRG cells at St. 20 using HNK-1/bromodeoxyuridine double-stain immunocytochemistry. The percentage of cells in S-phase at both brachial and cervical levels of the neuraxis in newly formed ganglia was observed to be the same. These two lines of evidence strongly argue that the initial difference in size between DRG at different axial levels is not intrinsic, but rather is imposed by the mesodermal microenvironment in which they develop, as is the case for DRG segmentation. This is in contrast to what may occur in the hindbrain, where determination of rhombomere identity and neural crest segmentation are thought to be intrisic to the nervous system.