Effects of hydroxyurea during final neuronal DNA synthesis in dorsal root ganglia of rats

Abstract

The sequence of final neuronal DNA synthesis was investigated in developing lumbar dorsal root ganglia of rats. Patterns of final division were compared to permanent neuronal deficiencies produced by single doses of hydroxyurea (HU), a specific cytotoxic inhibitor of DNA synthesis. The purpose was to discern increased susceptibility of terminal cell cycles in order to evaluate possible phenotypic or mitotic commitments responsible for cessation of DNA synthesis. The normal period of final DNA synthesis was found to occur primarily on gestation Days 12, 13, and early 14. Precursors of larger (A cells) and smaller (B cells) neurons are generated in sequence, suggesting the presence of phenotypic commitments during terminal division. When HU was administered during the period of final DNA synthesis, severe neuronal depletions and altered phenotypic proportions were observed postnatally. With HU on Day 13, total neuronal numbers were reduced by an average of 62% and deficiencies were confined primarily to neurons originating at or near the time of treatment. Given 12 hr later (Day 13.5), HU produced a 48% depletion involving neurons of smaller diameters. With treatment on Day 14, some ganglia appeared normal histologically but quantitation revealed an average 21% numerical deficiency involving the smallest neuronal phenotypes. Later treatments did not appear to affect ganglion morphology even though other defects (primarily growth retardation and gait abnormalities) continued to occur. Earlier treatments, given during terminal division of large neurons on Day 12, produced resorption or early postnatal death. The results suggest the emergence of phenotypic commitments in final cell cycles which restrict the probability of continued DNA synthesis and, thus, the probability of regeneration.