Quantitation of mRNAs for α1-acid glycoprotein and for serum albumin in livers of normal, stressed, fasted, and refed rats. [125I or 131I radioimmunoassay for protein products of specific mRNA activity]
- Univ. of Rochester, NY (United States)
A new procedure for determining the relative levels of a specific mRNA species was developed and applied to mRNA for rat serum albumin (RSA) and α1-AGP) in rat liver. The method is a radioimmunoassay (125In or 131I) for the completed protein, but which also detects antigenic determinants in nascent polypeptide chains on plysomes synthesizing the specific protein. Results show that 24 hs after stressing the rat by turpentine injection the total number of polysomes per mg DNA has increased by 20 to 25%; however, the number of RSA synthesizing polysomes per mg DNA has decreased slightly. In rats fasted for 6 days, the number of RSA synthesizing polysomes per mg polysomal RNA is only slightly below normal, but the total number of RSA synthesizing polysomes per mg DNA has decreased by 40%. Again, it is seen that RSA mRNA levels do not decrease as sharply as the rate of RSA synthesis. Twelve hours after refeeding the rats, the number of RSA synthesizing polysomes begins to increase, reaching a peak two to three times normal levels 24 to 48 hours after commencement of refeeding. During the first 24 hs after turpentine injection, there is a linear increase in the number of α1-AGP synthesizing polysomes. The increase is smaller during the next 24 hs and there is a small decrease between 48 and 72 hs. The serum concentrations of α1-AGP following turpentine treatment reflect these changes in polysome levels. It was not possible to compare the number of α1-AGP synthesizing polysomes in livers of normal, fasted, and refed rats because the levels detected were only slightly higher than those seen in rat and rat kidney polysome controls. This background activity must be eliminated before the technique can be applied to quantitating mRNA for proteins synthesized in very small quantities. This technique offers several advantages over other procedures commonly used to quantitate mRNA. (ERB)
- Research Organization:
- Univ. of Rochester, NY (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- EY-76-C-02-3490
- OSTI ID:
- 6453913
- Report Number(s):
- UR-3490-1495
- Resource Relation:
- Other Information: Thesis. Portions of document are illegible
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ALBUMINS
RADIOIMMUNOASSAY
BIOLOGICAL STRESS
BIOLOGICAL EFFECTS
FASTING
GLUCOPROTEINS
LIVER
MESSENGER-RNA
MEASURING METHODS
EXPERIMENTAL DATA
IODINE 125
IODINE 131
ISOLATED VALUES
QUANTITY RATIO
RATS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOHYDRATES
DATA
DATA FORMS
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
GLANDS
INFORMATION
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
NUCLEI
NUCLEIC ACIDS
NUMERICAL DATA
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RADIOASSAY
RADIOISOTOPES
RNA
RODENTS
SACCHARIDES
TRACER TECHNIQUES
VERTEBRATES
550601* - Medicine- Unsealed Radionuclides in Diagnostics
550201 - Biochemistry- Tracer Techniques
550901 - Pathology- Tracer Techniques