PubMed HealthSearch

Biomedical subjects

A M Giuffrida

Publications and source records attributed to A M Giuffrida.

7 recordsLinked to original sources

Macromolecular synthesis in mitochondria isolated from different regions of developing rat brain.

DNA, RNA, and protein synthesis in mitochondria isolated from cerebral hemispheres, brain stem, and cerebellum of 10- and 30-day-old rats was measured. Synthesis of different macromolecules was affected by the respective mitochondrial specific inhibitors, showing a good level of purity of mitochondrial preparations. DNA and protein synthesis in 10-day-old rats was about 70% higher than in 30-day-old animals. In contrast, RNA synthesis did not decrease with age in all the regions examined.

Aging

Mitochondrial DNA, RNA, and protein synthesis in different regions of developing rat brain.

In vivo and in vitro (tissue slices) incorporation of labeled precursors into DNA, RNA, and proteins was measured in mitochondria obtained from cerebral hemispheres, cerebellum, and brain stem of rats at different days of postnatal development. To compare the synthesis of macromolecules in mitochondria with that in other subcellular fractions, the incorporation of labeled precursors into DNA, RNA, and proteins extracted from nuclei and into RNA and proteins extracted from microsomes and cytoplasmic soluble fractions was also measured. The results obtained showed that the incorporation of [3H]thymidine into DNA and of [14C]leucine into proteins of nuclei and mitochondria from the various brain regions examined decreased during postnatal development; however, at 30 days of age the specific radioactivity of mitochondrial DNA was higher than that of nuclear DNA. [3H]Uridine incorporation into RNA decreased from 10 to 30 days of age in nuclei while in mitochondria it was quite similar at both ages. This result may be due to a faster turnover of mitochondrial RNA compared to that of mitochondrial DNA and proteins. The results obtained suggest an active biosynthesis of macromolecules in brain mitochondria and might indicate an intense biogenesis of these organelles in rat brain during postnatal development.

Animals

Biosynthesis of DNA and RNA in neuronal and glial cells from various regions of developing rat brain.

Slices of cerebral hemispheres, brain stem, and cerebellum from rats 5-30 days old were used for in vitro incorporation of [methyl-3H]thymidine and [6-14C]orotic acid into DNA and RNA, respectively. The rates of DNA and RNA synthesis decreased markedly during development, with the most marked decrease observed for DNA. The different brain regions showed specific patterns of decline of DNA and RNA synthesis. Following incubation of slices, the tissues were fractionated to obtain fractions enriched in neuronal cells and in glial cells. In cerebellum, the granule neurons were separated from the Purkinje neurons. The glial:neuronal ratio of DNA specific activity was different in the three regions examined: in cortex it decreased from 6 at 10 days to 3 at 20-30 days; in brain stem it was 3 throughout 10-30 days; in the cerebellum (glia:granule neuron ratio) it was also 3 at 30 days but only 0.3 at 10 days. Concerning the RNA incorporation, small differences were found between neuronal and glial cells.

Age Factors

Nucleolar and nucleoplasmic RNA polymerase activity in different regions of rat brain during postnatal development.

RNA polymerase activities of whole nuclei, of isolated and purified nucleoli and of the nucleoplasmic fractions obtained from cerebral hemispheres, cerebellum and brain stem of rat at different days of postnatal development have been determined. In the whole nuclei the fraction of RNA polymerase which is sensitive to alpha-amanitin, is strongly affected by salt concentration; at low ionic strength most of the activity is resistant to the drug while at high ionic strength the enzymatic activity shows a greater sensitivity to the drug. In isolated nucleoli RNA synthesis is not inhibited at all by alpha-amanitin. The biosynthesis of RNA, at low ionic strength, is inhibited by low doses of actinomycin D, whereas at high ionic strength it is remarkably inhibited only by higher doses of the drug. The sensitivity of the reaction to alpha-amanitin and actinomycin D provide good evidence that UTP or GTP incorporation into RNA in purified nuclei and nucleoli, is dependent on RNA polymerases acting on DNA template and is not dependent on homopolymer formation. These results show that in the whole brain nuclei at low ionic strength there is a preferential synthesis of rRNA, whereas at high ionic strength the synthesis of heterogenous RNA predominates. In isolated nucleoli the synthesis of RNA is restricted to rRNA.

Aging