The Relation Between the Intracellular Ribonucleic Acid Distribution and Amino Acid Incorporation in the Liver of the Developing Chick Embryo

Duck-Chong, C G; Pollak, John K.; North, Robert J.

doi:10.1083/jcb.20.1.25

Cited by 16 publications

(6 citation statements)

References 17 publications

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“…To isolate the DNA component we measured RNase treated cells, and determined a G1 DNA mass of 7.4 pg. These results are consistent with the published numbers for the genomic DNA and the RNA/DNA ratio measured in similar CHO cells using flow cytometry and Feulgen densitometry (16, 21 – 23, 31, 42 – 43). …”

Section: Discussionsupporting

confidence: 91%

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Deep ultraviolet mapping of intracellular protein and nucleic acid in femtograms per pixel

et al. 2011

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By using imaging spectrophotometry with paired images in the 200- to 280-nm wavelength range, we have directly mapped intracellular nucleic acid and protein distributions across a population of Chinese hamster ovary (CHO-K1) cells. A broadband 100× objective with a numerical aperture of 1.2NA (glycerin immersion) and a novel laser-induced-plasma point source generated high-contrast images with short (~100 ms) exposures and a lateral resolution nearing 200 nm that easily resolves internal organelles. In a population of 420 CHO-K1 cells and 477 nuclei, we found a G1 whole-cell nucleic acid peak at 26.6 pg, a nuclear-isolated total nucleic acid peak at 11.4 pg, and, as inferred by RNase treatment, a G1 total DNA mass of 7.4 pg. At the G1 peak we found a whole-cell protein mass of 95.6 pg, and a nuclear-isolated protein mass of 39.3 pg. An algorithm for protein quantification that senses peptide-bond (220-nm) absorbance was found to have a higher signal-to-noise ratio and to provide more reliable nucleic acid and protein determinations when compared to more classical 280-nm/260-nm algorithms when used for intracellular mass mapping. Using simultaneous imaging with common nuclear stains (Hoechst 33342, Syto-14, and Sytox Orange), we have compared staining patterns to deep-UV images of condensed chromatin and have confirmed bias of these common nuclear stains related to nuclear packaging. The approach allows absolute mass measurements with no special sample preparation or staining. It can be used in conjunction with normal fluorescence microscopy and with relatively modest modification of the microscope.

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Section: Discussionsupporting

confidence: 91%

“…While the qualitative features of the histograms are quite similar to cytometry data for both algorithms (20 – 21, 34), some problems in terms of statistical broadening and absolute calibration appear for the classical 260/280-nm data reduction. For the nucleic acid by this classical algorithm, there is a well-shaped histogram (Fig.…”

Section: Resultsmentioning

confidence: 68%

Deep ultraviolet mapping of intracellular protein and nucleic acid in femtograms per pixel

et al. 2011

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“…Nuclei (1) and mitochondria (55, 61) possess recognized mechanisms for production of proteins . When expressed as grain concentration per unit area (Table I), the 4 min values for nuclei and mitochondria are below the average for liver, in accord with the lower incorporation of labeled amino acids into nuclei and mitochondria found when these components were isolated after labeling (14,36,54) .…”

Section: Discussionmentioning

confidence: 61%

Electron Microscopic Radioautographic Detection of Sites of Protein Synthesis and Migration in Liver

Ashley

Peters

1969

The Journal of Cell Biology

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The sites of synthesis of proteins and their subsequent migration in rat liver have been studied during a 75 min period after labeling of liver-slice proteins by exposure to leucine-H3 for 2 min. Incorporation of the label into protein began after 1 min and was maximal by 4 min. Electron microscopic radioautography showed that synthesis of proteins in hepatocytes occurs mainly on ribosomes, particularly those in rough endoplasmic reticulum and, to some extent, in nuclei and mitochondria. Most of the newly formed proteins leave the endoplasmic reticulum in the course of 40 min, and concurrently labeled proteins appear in Golgi bodies, smooth membranes, microbodies, and lysosomes. A likely pathway for the secretion of some or all plasma proteins is from typical rough endoplasmic reticulum to a zone of reticulum which is partially coated with ribosomes, to the Golgi apparatus, and thence to the cell periphery. The formation of protein by reticuloendothelial cells was measured and found to be about 5% of the total protein formed by the liver.

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“…Protein synthesis was 20 times faster in the fetal kidney than in the adult kidney ( Fig. 4 a); Duck-Chong et al (28) found a difference of a similar magnitude between fetal and adult liver in the chick. The apparent 100-fold difference between fetal and adult kidney shown in Fig.…”

Section: Protein Synthesismentioning

confidence: 82%

Development of the Metanephric Kidney

Priestley

Malt

1968

The Journal of Cell Biology

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The metanephric kidney was studied in fetal and older mice beginning at 16 days after mating of the parents. Polyribosomes from fetal kidneys labeled in vitro with 4C-labeled amino acids had 10-20 times more acid-precipitable radioactivity associated with them than polysomes from adult kidneys similarly labeled. Between 3 and 6 days after birth the rate incorporation of labeled amino acids by polyribosomes from neonatal kidneys declined sharply to only twice the value found for adult kidneys. There was no change in the shape of the polyribosome profile with increasing age, but before birth few, if any, ribosomes were bound to membranes compared with 20% 2 days after birth and between 20 and 30% in the adult. Total protein represented less than 10% of the wet weight in the fetal kidney but increased to 17 % of the wet weight in the adult kidney. There was a steady decline in the concentration of RNA and DNA with respect to dry weight throughout kidney development. DNA concentration declined more rapidly than RNA concentration, so that the milligram to milligram ratio of RNA to DNA increased. In males the RNA/DNA ratio was stable at 1.3 at 40 days after birth; but in females the decline in DNA concentration was more protracted, and at 200 days after birth the RNA/DNA ratio was only 0.99. Thus, total nucleic acids show only gradual changes in concentration throughout development of the kidney, but a sharp change in the synthetic activity of the ribosomes and in their binding to membranes occurs in kidneys scon after birth.The biochemistry of development has been studied most extensively in echinoderm and amphibian eggs, in which changes in nucleic acid and protein metabolism can be related to distinct phases of morphogenesis. The heterogeneity and the complex stimuli governing the growth of most mammalian organs make them less convenient for biochemical investigation. Nonetheless, the synthesis of ribosomes, ribosomal RNA (rRNA), and messenger RNA (mRNA) during compensatory growth can be analyzed in an organ as heterogeneous as the kidney (1-3).In this paper we describe the patterns of protein and nucleic acid synthesis in the metanephros of the mouse during rapid normal growth from fetal to adult life. An intense synthesis of protein on free, i.e. not membrane-bound, polysomes during late fetal and early neonatal life ends abruptly between 3 and 6 days after birth; at the same time, membrane-bound ribosomes increase at the expense of free ribosomes. Thereafter, protein accumulates with little increase in DNA and with production of RNA proportionate only to maintaining a stable concentration of RNA in the organ. A preliminary statement of some of these findings has appeared (4). MATERIALS AND METHODS KidneysMice used in this study were Ham/ICR albinos (Charles River Breeding Laboratories, North Wilmington, Mass.). For experiments with fetal and juvenile kidneys, a single group of females were mated 42 days after their birth. The appearance of a vaginal plug in the mother was regarded as the first day of fet...

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The Relation Between the Intracellular Ribonucleic Acid Distribution and Amino Acid Incorporation in the Liver of the Developing Chick Embryo

Cited by 16 publications

References 17 publications

Deep ultraviolet mapping of intracellular protein and nucleic acid in femtograms per pixel

Deep ultraviolet mapping of intracellular protein and nucleic acid in femtograms per pixel

Electron Microscopic Radioautographic Detection of Sites of Protein Synthesis and Migration in Liver

Development of the Metanephric Kidney

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