Movement of [U-14C]glucose Carbon into and Subsequent Release from lipids and High-Molecular-Weight Constituents of Rat Brain, Liver, and Heart In Vivo

1972 ◽  
Vol 50 (1) ◽  
pp. 91-105 ◽  
Author(s):  
R. Vrba ◽  
Anna Winter

After subcutaneous injection of [U-14C]glucose into rats the amount of 14C incorporated in vivo into proteins was always higher than into lipids in brain, liver, and heart. The specific radioactivity of brain proteins was higher than those of liver and heart. Blood-brain comparisons show that protein carbon is derived continuously from glucose in the brain in situ and not as a result of deposition of amino acids or proteins from the circulation. Seventy-two percent of 14C in purified brain protein fractions was found in the amino acids of the hydrolysates of these fractions, mainly in alanine, glutamic, and aspartic acids. Maximum labelling was reached about 4 h after injection of [U-14C]glucose. Elimination of 14C from three classes of brain proteins (high-speed supernatant, particulate deoxycholate extractable, and residual) followed a biphasic time-course. The extent of labelling of, and the rate of elimination of 14C from, the three classes of rat brain proteins were very similar. The fate of 14C in the other investigated tissue fractions of brain, liver, and heart was compared with the fate of 14C in brain proteins.The results lend further support to the previously published suggestion that: (a) brain does not contain appreciable amounts of metabolically inert proteins or of proteins with turnover rates significantly higher than the mean for the bulk of brain proteins; (b) glucose carbon participates at a different rate and to a different extent in the metabolism of high-molecular-weight constituents of brain as compared to liver, heart, and plasma proteins; (c) the continuous conversion of glucose carbon into protein is an important part of the maintenance of the homeostasis of tissue proteins in vivo.

1970 ◽  
Vol 116 (3) ◽  
pp. 349-355 ◽  
Author(s):  
R. F. Peters ◽  
M. C. Richardson ◽  
Margaret Small ◽  
A. M. White

1. The powerful anti-inflammatory glucocorticoid triamcinolone acetonide, administered to rats at 20 and 2.5mg/kg, leads to a decrease in the incorporation in vivo of [3H]uridine and [32P]orthophosphate into hind-limb skeletal muscle. 2. At the higher dose, this decrease in the rate of incorporation of precursors into RNA precedes a decrease in the incorporating ability of muscle ribosomes, which commences about 4–5h after drug administration, but is unaccompanied by any changes in the concentration of tissue ATP or free amino acids. 3. The ribosomal dysfunction extends to polyribosomes, which can only be successfully isolated from the muscle of triamcinolone-treated animals after the addition of α-amylase to the tissue homogenate to remove glycogen. 4. The specific radioactivity of muscle protein labelled in vivo with 14C-labelled amino acids does not decrease progressively after triamcinolone administration. After 2h there is an apparent stimulation of incorporation which leads to an overall discrepancy between measurements of protein-synthetic activity made in vivo and in vitro. 5. There is a significant increase in muscle-glycogen concentration between 8 and 12h after the administration of triamcinolone acetonide (20mg/kg), although a significant decrease occurs after 4h. The fall in glycogen concentration may be due to a decrease in the rate of synthesis of protein essential for glucose uptake into the tissues. 6. As judged by (a) incorporation of 14C-labelled amino acids into protein, (b) [3H]uridine and [32P]-orthophosphate incorporation into RNA, (c) the rate of induction of tryptophan pyrrolase and (d) changes in the pool sizes of taurine and tryptophan, the responses in liver followed the same time-course as those in muscle after administration of the drug.


1978 ◽  
Vol 176 (2) ◽  
pp. 511-521 ◽  
Author(s):  
C Hall ◽  
L Lim

The regional metabolism of high-molecular-weight RNA in the developing female rat brain was investigated after the intracranial injection of [32P]P1. The synthesis of polyadenylated RNA relative to high-molecular-weight RNA was determined after oligo(dT)-cellulose chromatography of total cellular high-molecular-weight RNA labelled after 4h. In both hypothalamus and cortex this synthesis was significantly higher during the first 10 days post partum than at subsequent ages. In both regions apparently more mRNA is synthesized in the young. The ratio of the specific radioactivity of cytoplasmic high-molecular-weight RNA relative to that of the nucleus, measured after a 48 h period of labelling, was considered to be an index of the nucleocytoplasmic transport of newly synthesized RNA [Berthold & Lim (1976) Biochem. J. 154, 529–539]. In the cortex, nucleo-cytoplasmic RNA transport in rats aged up to 20 days was significantly higher than in older rats, with the maximal value being attained between 16 and 19 days post partum. In contrast, in the hypothalamus, nucleo-cytoplasmic transport of RNA was low during the neonatal period and comparable with that of the mature animal. However, there were two periods of increased transport at later stages of development, the first between 15 and 19 days post partum and the second between 25 and 29 days post partum. These prepubertal changes in the nucleo-cytoplasmic transport of RNA in the female hypothalamus during weeks 3 and 4 post partum are coincident with other reported changes occurring during sexual differentiation. Differences in the timing of the maturational changes of the two brain regions thus appear to be reflected in developmental changes in RNA transport.


1971 ◽  
Vol 122 (2) ◽  
pp. 135-138 ◽  
Author(s):  
Jill E. Cremer

The metabolism of ketone bodies by rat brain was studied in vivo. Rats starved for 48h were given either d-β-hydroxy[3-14C]butyrate or [3-14C]acetoacetate by intravenous injection and killed after 3 or 10min. Total radioactivity in the acid-soluble material of the brain and the specific radioactivities of the brain amino acids glutamate, glutamine, aspartate and γ-aminobutyrate were determined. A group of fed animals were also given d-β-hydroxy[3-14C]butyrate. In the brains of all animals 14C was present in the acid-soluble material and the specific radioactivity of glutamate was greater than that of glutamine.


2021 ◽  
Author(s):  
Nan Zheng ◽  
Xiahui Li ◽  
Shangwei Huangfu ◽  
Kangkai Xia ◽  
Ruofei Yue ◽  
...  

A linear poly-porphyrin with high Mw and conjugated by PEG and acetazolamide was developed with enhanced singlet oxygen quantum yield, improved photo-toxicity and excellent in vivo photodynamic therapy.


1984 ◽  
Vol 99 (4) ◽  
pp. 1372-1378 ◽  
Author(s):  
K A Resing ◽  
K A Walsh ◽  
B A Dale

A major event in the keratinization of epidermis is the production of the histidine-rich protein filaggrin (26,000 mol wt) from its high molecular weight (greater than 350,000) phosphorylated precursor (profilaggrin). We have identified two nonphosphorylated intermediates (60,000 and 90,000 mol wt) in NaSCN extracts of epidermis from C57/Bl6 mice by in vivo pulse-chase studies. Results of peptide mapping using a two-dimensional technique suggest that these intermediates consist of either two or three copies of filaggrin domains. Each of the intermediates has been purified. The ratios of amino acids in the purified components are unusual and essentially identical. The data are discussed in terms of a precursor containing tandem repeats of similar domains. In vivo pulse-chase experiments demonstrate that the processing of the high molecular weight phosphorylated precursor involves dephosphorylation and proteolytic steps through three-domain and two-domain intermediates to filaggrin. These processing steps appear to occur as the cell goes through the transition cell stage to form a cornified cell.


1985 ◽  
Vol 248 (4) ◽  
pp. R391-R399 ◽  
Author(s):  
J. Katz

A mitochondrial model of gluconeogenesis and the tricarboxylic acid cycle, where pyruvate is metabolized via pyruvate carboxylase and pyruvate dehydrogenase, and pyruvate kinase is examined. The effect of the rate of tricarboxylic acid flux and the rates of the three reactions of pyruvate metabolism on the labeling patterns from [14C]pyruvate and [24C]acetate are analyzed. Expressions describing the specific radioactivities and 14C distribution in glucose as a function of these rates are derived. Specific radioactivities and isotopic patterns depend markedly on the ratio of the rates of pyruvate carboxylation and decarboxylation to the rate of citrate synthesis, but the effect of phosphoenolpyruvate hydrolysis is minor. The effects of these rates on 1) specific radioactivity of phosphoenolpyruvate, 2) labeling pattern in glucose, and 3) contribution of pyruvate, acetyl-coenzyme A, and CO2 to glucose carbon are illustrated. To determine the contribution of lactate or alanine to gluconeogenesis, experiments with two compounds labeled in different carbons are required. Methods in current use to correct for the dilution of 14C in gluconeogenesis from [14C]pyruvate are shown to be erroneous. The experimental design and techniques to determine gluconeogenesis from 14C-labeled precursors are presented and illustrated with numerical examples.


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