On using cis-Pt (II)-uracil as a stain for nucleic acids in brain slices and subcellular fractions

1976 ◽  
Vol 49 (3) ◽  
pp. 253-261 ◽  
Author(s):  
Joseph A. Babitch ◽  
Donald L. Helseth ◽  
Tien-Cheng Chiu
Keyword(s):  
Nucleic Acids ◽  
Brain Slices ◽  
Subcellular Fractions

scholarly journals Choice of precursors for the measurement of protein turnover by the double-isotope method. Application to the study of mitochondrial proteins

1978 ◽  
Vol 176 (3) ◽  
pp. 919-926 ◽  
Author(s):  
R J Burgess ◽  
J H Walker ◽  
R J Mayer
Keyword(s):  
Solvent Extraction ◽  
Nucleic Acids ◽  
Protein Turnover ◽  
Isotope Ratio ◽  
Subcellular Fractions ◽  
Turnover Rates ◽  
Extraction Procedures ◽  
Mitochondrial Fractions ◽  
Double Isotope ◽  
Submitochondrial Fractions

1. The double-isotope concept [Arias, Doyle & Schimke (1969) J. Biol. Chem. 224, 3303–3315] for the measurement of protein turnover was used to estimate the turnover of proteins in subcellular and submitochondrial fractions prepared from rat liver. 2. Double-isotope experiments with [3H]leucine as first precursor and [14C]leucine as second precursor were used to measure the turnover rates of proteins in subcellular and submitochondrial fractions. Solvent extraction procedures designed to remove lipids and nucleic acids from trichloroacetic acid precipitates only changed the isotope ratio of the microsomal fraction. It was not possible to measure turnover of proteins in mitochondrial and submitochondrial fractions with these precursors. 3. Double-isotope experiments were designed to minimize first-precursor reutilization by employing NaH14CO3. [3H]Arginine was used as second precursor. The turnover rates of protein in subcellular and submitochondrial fractions was measured. Solvent extraction procedures designed to remove lipids and nucleic acids showed changes in the isotope ratio for all subcellular fractions, especially in microsomal and detergent-soluble mitochondrial fractions. Isotope ratios of precipitates after solvent extraction indicate that, whereas considerable heterogeneity exists for the average rates of protein turnover in subcellular fractions, little heterogeneity is observed in the average rates of protein turnover in submitochondrial fractions.

scholarly journals The metabolism of phospholipids in mouse brain slices

1966 ◽  
Vol 101 (3) ◽  
pp. 674-679 ◽  
Author(s):  
PA Clayton ◽  
CE Rowe
Keyword(s):  
Phosphatidic Acid ◽  
Mouse Brain ◽  
Grey Matter ◽  
Microsomal Fraction ◽  
Brain Slices ◽  
Mitochondrial Fraction ◽  
Subcellular Fractions ◽  
The Other ◽  
Metabolic Inhibitors ◽  
Radioactive Labelling

1. Slices of mouse brain grey matter were incubated with [(32)P]phosphate and [1-(14)C]acetate. Doubly labelled phospholipids were extracted from subcellular fractions prepared from the slices in a mixture of metabolic inhibitors, under conditions where there was negligible change in radioactive labelling during the preparation. Two tissue fractions were studied in detail; one contained a high proportion of mitochondria and the other was mainly microsomal. 2. In all tissue fractions the highest incorporations of both [(32)P]phosphate and [1-(14)C]acetate occurred into phosphatidylcholine. 3. After incubation for 1hr., the (32)P/(14)C ratios for phosphatidylcholine, phosphatidylethanolamine and phosphatidic acid in the mitochondrial fraction were similar to those in the microsomal fraction. 4. The (32)P/(14)C ratios were similar in phosphatidylcholine and phosphatidylethanolamine and much lower than those in phosphatidic acid and phosphatidylinositol.

scholarly journals Incorporation of radioactive phosphate into the nucleic acids of brain slices

1953 ◽  
Vol 55 (2) ◽  
pp. 193-200 ◽  
Author(s):  
H. A. Deluca ◽  
R. J. Rossiter ◽  
K. P. Strickland
Keyword(s):  
Nucleic Acids ◽  
Brain Slices

Nucleic Acids From Subcellular Fractions of N-Nitrosodiethylamine-Induced Hepatoma in Mice

1976 ◽  
Vol 57 (1) ◽  
pp. 23-31 ◽  
Author(s):  
Jacob D. Shaposhnikov ◽  
Wolf N. Shalumovich ◽  
Oleg I. Kisselev ◽  
Vladimir S. Gaitskhoki ◽  
Kazymir M. Pozharisski
Keyword(s):  
Nucleic Acids ◽  
Subcellular Fractions

Electron Microscopy of Nucleic Acids

1974 ◽  
Vol 32 ◽  
pp. 302-303
Author(s):  
Norman Davidson
Keyword(s):  
Electron Microscopy ◽  
Nucleic Acids ◽  
Basic Protein ◽  
Molecular Biologist ◽  
Topological Properties ◽  
Protein Film ◽  
Polynucleotide Chain

The basic protein film technique for mounting nucleic acids for electron microscopy has proven to be a general and powerful tool for the working molecular biologist in characterizing different nucleic acids. It i s possible to measure molecular lengths of duplex and single-stranded DNAs and RNAs. In particular, it is thus possible to as certain whether or not the nucleic acids extracted from a particular source are or are not homogeneous in length. The topological properties of the polynucleotide chain (linear or circular, relaxed or supercoiled circles, interlocked circles, etc. ) can also be as certained.

mRNA localization by Electron microscopic in situ hybridization

1991 ◽  
Vol 49 ◽  
pp. 430-431
Author(s):  
J. A. Pollock ◽  
M. Martone ◽  
T. Deerinck ◽  
M. H. Ellisman
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Nucleic Acids ◽  
Recombinant Dna ◽  
Light And Electron Microscopy ◽  
Electron Microscopic ◽  
High Voltage Electron Microscopy ◽  
Functional Sites ◽  
Voltage Electron

Localization of specific proteins in cells by both light and electron microscopy has been facilitate by the availability of antibodies that recognize unique features of these proteins. High resolution localization studies conducted over the last 25 years have allowed biologists to study the synthesis, translocation and ultimate functional sites for many important classes of proteins. Recently, recombinant DNA techniques in molecular biology have allowed the production of specific probes for localization of nucleic acids by “in situ” hybridization. The availability of these probes potentially opens a new set of questions to experimental investigation regarding the subcellular distribution of specific DNA's and RNA's. Nucleic acids have a much lower “copy number” per cell than a typical protein, ranging from one copy to perhaps several thousand. Therefore, sensitive, high resolution techniques are required. There are several reasons why Intermediate Voltage Electron Microscopy (IVEM) and High Voltage Electron Microscopy (HVEM) are most useful for localization of nucleic acids in situ.

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