scholarly journals Influence of steroid-hormone-receptor-protein complexes on initiation of ribonucleic acid synthesis in liver nuclei isolated from rats of various ages

1981 ◽  
Vol 196 (1) ◽  
pp. 373-375 ◽  
Author(s):  
J K Miller ◽  
R Bolla

The effect of age on the induction of the initiation of RNA synthesis was investigated in liver nuclei isolated from adrenalectomized rats of various ages after binding of a dexamethasone-receptor-protein complex. Binding of this complex to nuclear chromatin resulted in increased initiation of nuclear RNA synthesis at all ages; however, an age-associated decline in the extent of this induction was observed. This suggests an age-related decrease of total rat liver nuclear RNA synthesis with a decreased response to glucocorticoid hormones.

1971 ◽  
Vol 49 (7) ◽  
pp. 853-862 ◽  
Author(s):  
Peter M. K. Ip ◽  
Maurice Brossard

RNA has been isolated and fractionated from rat liver nuclei by a sequence of salt extractions into four subfractions: namely nucleoplasmic, deoxyribonucleoprotein-associated, ribonucleoprotein-associated, and nucleolar fractions.Study of 14C-methyl-methionine labeling indicates that the nucleolar RNA fraction isolated by the present procedure is, in fact, of nucleolar origin while the other three fractions are essentially of extranucleolar origin.The effects of growth hormone and hydrocortisone on the nuclear RNA synthesis have been further studied using the present isolation and double-labeling techniques. Both hormones cause an increase in the incorporation of labeled orotic acid into all types of RNA in all four nuclear subfractions. However, the stimulatory effect of growth hormone is found mainly in the 18–28 S and 45–60 S regions of the nucleolar fraction and in regions of the entire gradient of the nucleoplasmic fraction, whereas the stimulatory effect of hydrocortisone is localized mainly in the 10–28 S regions of the nucleolar fraction and in the 4 S and 10–18 S regions of the ribonucleoprotein fraction.The present report suggests that there are qualitative as well as quantitative differences in the action of growth hormone and hydrocortisone on the synthesis of RNA in liver nucleus. The mode of action of the two hormones is discussed.


1979 ◽  
Vol 184 (3) ◽  
pp. 669-674 ◽  
Author(s):  
R Bolla ◽  
W D Denckla

Changes in RNA synthesis in liver nuclei were observed at different ages and after hypophysectomy and hormone replacement in female Sprague-Dawley rats. As determined by the incorporation of [3H]UMP into an acid-insoluble product, RNA synthesis decreased by about 75% in intact rats from 6 months to 24 months of age. This decline with age was not observed in liver nuclei from 24-month-old rats that had been hypophysectomized at 12 months and maintained on a minimal hormone-replacement therapy. Thyroid hormones and somatotropin (growth hormone) had an additive effect on RNA synthesis in liver nuclei from these hypophysectomized rats. The same hormones had no significant effect on intact, age-matched rats. With advancing age, nuclei of intact rats had an increase in the pool of free RNA polymerase and an apparent decrease in the enzyme activity bound to nuclear chromatin. There was no change in total enzyme with age. In hypophysectomized, hormone-treated rats, free RNA polymerase activity decreased and chromatin-bound activity increased. There was no difference in total nuclear RNA polymerase activity between operated or intact rats. However, the ratio of the bound to the free activity was different. These results suggest that the ability of RNA polymerase to bind to chromatin may be involved in the age-related decrease in liver nuclear RNA synthesis of intact rats.


1980 ◽  
Vol 188 (1) ◽  
pp. 55-60 ◽  
Author(s):  
J K Miller ◽  
R Bolla ◽  
W D Denckla

Initiation of RNA synthesis was studied in an attempt to determine a possible molecular mechanism for age-related biochemical and physiological changes. Initiation of RNA synthesis was determined by incorporation of [gamma-32 P]ATP and of [gamma-32P]GTP into an acid-insoluble product by intact nuclei isolated from livers of Sprague-Dawley CD-strain rats of various ages. When the rats were grouped into young (0.75-9 months) and old (12-30 months) rats, a significant decrease (P less than or equal to 0.001) in incorporation of initiating nucleotides was observed. The rat population was divided into five age groups (0.75-3 months, 4-9 months, 12-18 months, 19-23 months and 30 months) for further analysis of the effect of age on the initiation of RNA synthesis. Analysis of data from these groups indicated a significant trend for an age-related decrease in RNA-synthesis initiation (correlation coefficient = 0.94). Long-term hypophysectomy coupled with minimal hormone-replacement therapy was shown to have a significant effect on the reversal of the age-related decrease in initiation of RNA synthesis. It was observed that initiation of RNA synthesis in nuclei from 19-month-old rats, hypophysectomized at 12 months of age, was closest to that in 3-month-old intact rats and was not significantly different from that in liver nuclei of 0.75-9-month-old intact rats.


1965 ◽  
Vol 49 (3_Suppl) ◽  
pp. S160 ◽  
Author(s):  
E. Stöcker ◽  
G. Dhom

1965 ◽  
Vol 27 (3) ◽  
pp. 545-550 ◽  
Author(s):  
A. Di Marco ◽  
R. Silvestrini ◽  
S. Di Marco ◽  
T. Dasdia

The effect has been studied of Actinomycin D, Daunomycin (Da.), and Da. N acetyl derivative on mitotic activity and on the nucleic acid synthesis of in vitro HeLa cell cultures. The experiments were carried out by means of the radioautographic technique using stripping films. The relative uptake of thymidine-H3 and uridine-H3 was determined by means of the reduced silver grain count present in the nuclei of controls and treated cells. The mitotic activity and thymidine incorporation were noticeably reduced by Daunomycin and Actinomycin, whereas both processes appeared less affected by Da. N acetyl derivative. As regards nuclear RNA synthesis, all three antibiotics at low doses chiefly inhibit nucleolar RNA synthesis. On the other hand, whilst Actinomycin at higher doses causes an almost total inhibition of the synthesis of the whole nuclear RNA, in Daunomycin- and Da. N acetyl derivative-treated cells extranucleolar RNA synthesis is less susceptible to inhibition.


1965 ◽  
Vol 26 (3) ◽  
pp. 937-958 ◽  
Author(s):  
Shuichi Karasaki

The site of H3-uridine incorporation and the fate of labeled RNA during early embryo-genesis of the newt Triturus pyrrhogaster were studied with electron microscopic autoradiography. Isolated ectodermal and mesodermal tissues from the embryos were treated in H3-uridine for 3 hours and cultured in cold solution for various periods before fixation with OsO4 and embedding in Epon. At the blastula stage, the only structural component of the nucleus seen in electron micrographs is a mass of chromatin fibrils. At the early gastrula stage, the primary nucleoli originate as small dense fibrous bodies within the chromatin material. These dense fibrous nucleoli enlarge during successive developmental stages by the acquisition of granular components 150 A in diameter, which form a layer around them. Simultaneously larger granules (300 to 500 A) appear in the chromatin, and they fill the interchromatin spaces by the tail bud stage. Autoradiographic examination has demonstrated that nuclear RNA synthesis takes place in both the nucleolus and the chromatin, with the former consistently showing more label per unit area than the latter. When changes in the distribution pattern of radioactivity were studied 3 to 24 hours after immersion in isotope at each developmental stage, the following results were obtained. Labeled RNA is first localized in the fibrous region of the nucleolus and in the peripheral region of chromatin material. After longer culture in non-radioactive medium, labeled materials also appear in the granular region of the nucleolus and in the interchromatin areas. Further incubation gives labeling in cytoplasm.


Nature ◽  
1972 ◽  
Vol 238 (5365) ◽  
pp. 457-459 ◽  
Author(s):  
R. M. BERNSTEIN ◽  
B. B. MUKHERJEE

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