Neuronal nuclei and glial nuclei from mammalian cerebral cortex. Nucleosome repeat lengths, DNA contents and H1 contents.
We have characterised the histone and DNA contents of neuronal and glial nuclei from ox cerebral cortex which have, respectively, repeat lengths of 162 base pairs and 201 base pairs. Although the neuronal population cannot be obtained completely free of glial nuclear contamination, the degree of contamination is easily determined by counting, and has been allowed for in all the methods used here. By diphenylamine assay and flow cytofluorometry we find that the DNA contents of both nuclear types are essentially equal, and equivalent to the diploid value, contrary to some reports. By quantification of the core histones in known numbers of nuclei with respect to an added external standard, we have shown that the ratio of core histone octamers in the two nuclear types, neuronal and glial, is the inverse of the ratio of repeat lengths. Thus the same proportion of DNA is associated with core histone octamers in the two nuclear types, most simply all of the DNA. By complete radiolabelling of the lysine side chains of the histones with methyl [1-3H]acetimidate we have determined the stoichiometry of H1 relative to the core histones. Neuronal nuclei have a low H1 content of 0.45 molecule H1/nucleosome on average; glial nuclei have the 'normal' 1 H1 molecule/nucleosome. In neuronal nuclei about half of the nucleosomes therefore probably lack H1. Whether there is any relation between the low H1 content and the short DNA repeat length of neuronal nuclei, on the one hand, and their high transcriptional capacity (at least when assayed in vitro), on the other, remains to be established.
D000818:Animals; D001482:Base Composition; D002417:Cattle; D002467:Cell Nucleus / Q000032:analysis*; D002540:Cerebral Cortex / Q000032:analysis* / Q000648:ultrastructure; D004247:DNA / Q000302:isolation & purification*; D004159:Diphenylamine; D005434:Flow Cytometry; D006657:Histones / Q000032:analysis*; D009457:Neuroglia / Q000032:analysis / Q000648:ultrastructure; D009474:Neurons / Q000032:analysis* / Q000648:ultrastructure; D009707:Nucleosomes / Q000032:analysis* / Q000648:ultrastructure; D051381:Rats; D012091:Repetitive Sequences, Nucleic Acid
D006657:Histones; D009707:Nucleosomes; D004247:DNA; D004159:Diphenylamine