A method for isolating pure and unaltered nuclei from rat brain by means of differential centrifugation is described. The isolated nuclei are further separated into discrete fractions of neuronal, astrocytic, and glial nuclei, with a yield amounting to 20 to 25 % of the DNA of the original homogenate. Both the morphology and size of the nuclei remained unchanged. Problems concerning the composition of the isolation media, the use of detergents, as well as those raised by density gradient centrifugation in sucrose, Ficoll, and Dextran are discussed. Some values for the density of each type of brain nuclei are suggested.

No abstract

Xenopus embryos at various developmental stages were incubated in the presence of labelled substrates and the ¹⁴CO₂ production determined. From the rates of oxidation of glucose labelled in positions 1 and 6 and from that of radioactive acetate, pyruvate and glutamate, it was concluded that the Embden-Meyerhof pathway and the Krebs cycle are functional during early embryogenesis, but that their relative participation in the metabolic processes is limited and increases from gastrulation onwards. Early development is characterized by the predominance of the pentose cycle and the glutamate-aspartate cycle. Furthermore, it was shown that glutamate may be the main energy source up to gastrulation.

In this study we investigated the variations of the maximal activities of the rate-controlling glycolytic enzymes (i.e., hexokinase, HK; phosphofructokinase, PFK; pyruvate kinase, PK) and of the pyruvate-dehydrogenase complex (PDHc) during the early embryogenesis of Xenopus laevis (from cleavage through hatching). All the enzymatic assays, using different coupled reactions, were performed spectrophotometrically on cytosolic and mitochondrial fractions. The maximal HK activity increases markedly from neurulation onwards, PFK activity presents a peak around gastrulation, PK activity remains relatively constant throughout the period studied and the highest PDHc activity is observed during cleavage. The specific activities display the same temporal pattern. Furthermore, in the sequence of reactions by which glucose is degraded to form acetyl-CoA, the maximal activities of PFK and PK are not limiting while those of HK and PDHc could be rate-limiting at relatively late developmental stages (hatching).

No abstract