DIFFERENTIAL APPEARANCE OF EXTENSIVELY PHOSPHORYLATED FORMS OF THE HIGH-MOLECULAR-WEIGHT NEUROFILAMENT PROTEIN IN REGIONS OF MOUSE-BRAIN DURING POSTNATAL-DEVELOPMENT

The appearance and accumulation of extensively phosphorylated forms of the high molecular weight neurofilament protein (H-phos) was studied in six regions of mouse brain during postnatal development by quantitative immunoblot analyses. H-phos (migrating at 200 kDa) was detected in brainstem, cerebellum, cortex and hippocampus as early as postnatal day 1. While NF-H levels increased dramatically during subsequent postnatal development in these regions, and reached levels similar to those observed in adult brain by postnatal day 14, quantitative differences were observed in both the rate and the extent of increase among individual regions. The most rapid accumulation of H-phos was observed in brainstem and cortex, where H-phos increased within the first postnatal week to levels comparable to those of adult brain. However, H-phos exhibited a slower developmental change in cerebellum, where the levels increased uniformly over the first two postnatal weeks. In hippocampus, the major increase in H-phos levels was delayed until the second postnatal week. In contrast to its early detection in the above regions, H-phos was not detected in immunoblot analyses of olfactory bulb or hypothalamus cytoskeletons at postnatal day 1, indicating that in these regions the accumulated levels of posttranslationally modified forms of this protein appeared relatively late. Furthermore, H-phos levels in hippocampus did not level off at postnatal day 14 and continued to increase until at least postnatal day 21. Immunoblot analyses of whole embryonic brain revealed the presence of H-phos as early as embryonic day 17, demonstrating that some mouse brain regions carry out extensive phosphorylation of NF-H during embryonic development. We conclude that extensive phosphorylation of NF-H is initiated relatively early during mouse brain development, but that the timing of expression of the NF-H polypeptide and extent of its phosphorylation varied significantly among brain regions.