Isolation and preservation of cell nuclei for studies on RNA polymerase activity

1970 ◽  
Vol 48 (5) ◽  
pp. 559-565 ◽  
Author(s):  
R. S. D. Read ◽  
C. M. Mauritzen
Keyword(s):  
Rna Polymerase ◽  
Mammalian Cell ◽  
Cell Types ◽  
Polymerase Activity ◽  
Cell Nuclei ◽  
Low Concentrations ◽  
Nonionic Detergent ◽  
Rna Polymerase Activity ◽  
Liver Nuclei ◽  
Nonionic Detergents

The suitability of saponin for the isolation and of glycerol for the preservation of mammalian cell nuclei has been investigated. The nonionic detergent saponin was found to be a useful cell lytic agent in a procedure for the isolation of nuclei from several mammalian cell types. The RNA polymerase activity of rat liver nuclei was not affected by treatment with saponin or with some other nonionic detergents that were tested. Low concentrations of ionic agents also did not affect the activity of the enzyme though higher concentrations caused lysis of the nuclei. The preservation of structure and enzyme activity in the isolated nuclei was achieved by storage at low temperature in a medium containing 70% glycerol together with a suitable concentration of a divalent metal and phosphate buffer.

Regulation of RNA polymerase activity in liver and brain cell nuclei by a cytoplasmic thyroxine modulator in rats of various ages

1993 ◽  
Vol 116 (1) ◽  
pp. 820-822
Author(s):  
Ya. Kh. Turakulov ◽  
S. N. Dalimova ◽  
I. R. Kamalieva ◽  
G. D. Umarova ◽  
B. A. Atakhanova
Keyword(s):  
Rna Polymerase ◽  
Polymerase Activity ◽  
Brain Cell ◽  
Cell Nuclei ◽  
Rna Polymerase Activity

scholarly journals Studies on ribonucleic acid and homopolyribonucleotide formation in neuronal, glial and liver nuclei

1970 ◽  
Vol 116 (4) ◽  
pp. 599-609 ◽  
Author(s):  
Takahiko Kato ◽  
Masanori Kurokawa
Keyword(s):  
White Matter ◽  
Guinea Pig ◽  
Rna Polymerase ◽  
Grey Matter ◽  
Polymerase Activity ◽  
Physical Relationship ◽  
Neuronal Nuclei ◽  
Cerebral Grey Matter ◽  
Rna Polymerase Activity ◽  
Liver Nuclei

1. Various types of nuclear preparations, with different ratios of neuronal to glial nuclei, were isolated from guinea-pig cerebral grey matter and ox cerebral grey matter and white matter. Conditions appropriate for the separate assay of RNA and poly A formation were described. Comparative rates of RNA and poly A formation were studied in cerebral and liver nuclei. 2. RNA polymerase activity per nucleus is higher in neuronal nuclei than in glial nuclei. In liver nuclei, the activity is much lower than in cerebral nuclei. The physical relationship between RNA polymerase and deoxyribonucleoprotein seems to differ in neuronal, glial and liver nuclei. 3. Poly A polymerase activity in liver nuclei is selectively activated by Mn2+ and inhibited by GTP, CTP and UTP. On a DNA basis, the activity in an aggregate enzyme is the same as in intact nuclei. Poly A polymerase activity per nucleus is much higher in liver nuclei than in neuronal nuclei. Glial nuclei show an intermediate activity. 4. It is suggested that, in neuronal nuclei, the synthesis of RNA is more prominent than that of poly A under conditions where both polymers are formed simultaneously. This contrasts with liver nuclei, where more poly A is made than RNA. 5. In neuronal nuclei, the rate of CTP incorporation is much higher than in glial and liver nuclei. This incorporation is most probably due to poly C synthesis.

scholarly journals Effect of hypophysectomy on liver nuclear ribonucleic acid synthesis in aging rats

1979 ◽  
Vol 184 (3) ◽  
pp. 669-674 ◽  
Author(s):  
R Bolla ◽  
W D Denckla
Keyword(s):  
Rna Polymerase ◽  
Rna Synthesis ◽  
Hormone Replacement ◽  
Polymerase Activity ◽  
Sprague Dawley ◽  
Age Related ◽  
Nuclear Rna ◽  
Rna Polymerase Activity ◽  
Liver Nuclei ◽  
Intact Rats

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.

The effect of malathion on RNA polymerase activity of cell nuclei and transcription products in lymphocyte culture

1986 ◽  
Vol 41 (2) ◽  
pp. 372-377 ◽  
Author(s):  
Hanna Wiszkowska ◽  
Irena Kulamowicz ◽  
Alicja Malinowska ◽  
Zofia Walter
Keyword(s):  
Rna Polymerase ◽  
Polymerase Activity ◽  
Lymphocyte Culture ◽  
Cell Nuclei ◽  
Rna Polymerase Activity
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