Biochemical characterization of nerve growth cones isolated from both fetal and neonatal rat forebrains: the growth cone particle fraction mainly consists of axonal growth cones in both stages.
Nerve growth cones are responsible for the exact pathway finding, and for the establishment of neurocytoarchitecture. To elucidate the developmental changes of biochemical characteristics of nerve growth cones, growth cone particle (GCP) fractions were isolated biochemically from embryonal day 17 (E17) rat forebrain and from postnatal day 5 (P5). There were no significant differences in protein phosphorylation pattern in a Ca(2+)-dependent manner between E17-GCP fraction and that of P5. As for the membrane lipid composition, molar ratios of cholesterol to total phospholipids were well conserved during these ages. The immunoreactivity to anti-synaptophysin monoclonal antibody as a marker of mature synaptic elements could not be detected either in E17-GCP or P5-GCP fractions. To exclude the possibility of the contamination of dendritic elements, RNA contents and immunoreactivity to anti-high molecular weight microtubule-associated protein 2 (MAP2) monoclonal antibody were examined. RNA contents of the GCP fractions were extremely low compared to those of other subcellular fractions both in E17 and P5. No immunoreactivities to anti-MAP2 antibody were observed in either GCP fraction. Our results suggest that the GCP fractions, isolated from forebrains of E17 to P5 rat, are free from the contamination of the synaptic elements, and that the GCP fractions are mainly composed of axonal growth cones.
Saito
S
S
Department of Molecular and Cellular Neurobiology, Gunma University School of Medicine, Japan.
Fujita
T
T
Komiya
Y
Y
Igarashi
M
M
D000375:Aging / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn / Q000502:physiology; D000911:Antibodies, Monoclonal / Q000276:immunology; D001369:Axons / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D002784:Cholesterol / Q000378:metabolism; D003712:Dendrites / Q000378:metabolism; D005260:Female; D005333:Fetus / Q000502:physiology; D008297:Male; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009474:Neurons / Q000378:metabolism / Q000502:physiology*; D010743:Phospholipids / Q000378:metabolism; D011247:Pregnancy; D016548:Prosencephalon / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000378:metabolism; D013569:Synapses / Q000502:physiology; D016708:Synaptophysin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D008869:Microtubule-Associated Proteins; D010743:Phospholipids; D016708:Synaptophysin; D012313:RNA; D002784:Cholesterol