Determination of DNA and RNA in homogenized cells and tissues by surface fluorometry

Karsten, Uwe; Wollenberger, A

doi:10.1016/0003-2697(72)90405-8

Cited by 171 publications

(39 citation statements)

References 20 publications

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“…This index would reach a value of 1 if all the protein kinase present were activated. The cAMP content of adrenal medullae was assayed as previously described (10), the monoamine oxidase (EC 1.4.3.4) activity according to Goridis and Neff (14), the DNA and RNA content according to Karsten and Wollenberger (15), and the protein content according to Lowry et al (16).…”

Section: Methodsmentioning

confidence: 99%

Activation and nuclear translocation of protein kinase during transsynaptic induction of tyrosine 3-monooxygenase.

Costa¹,

Kurosawa²,

Guidotti³

1976

Proc. Natl. Acad. Sci. U.S.A.

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The tyrosine-3-monooxygenase activity [Ltyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating); EC 1.14.16.2] of rat adrenal medulla is induced 20-24 hr after the injection of reserpine (16 tmol/kg intraperitoneally). This and other inducing stimuli increase the 3':5'-cyclic AMP (cAMP) content in the medulla for longer than 60 min and activate the cAMP-dependent protein kinase (ATP:protein phosphotransferase; EC 2.7.1.37) for several hours. Corticotropin (ACTH), dopamine, and propranolol do not induce the monooxygenase, but elicit an increase in the cAMP content of the medulla which fails to activate protein kinase and lasts less than 1 hr. A high-and low-molecular-weight protein kinase are separated by gel filtration from the 20,000 X g pellet extract of adrenal medulla homogenate. The activity of the low-molecular-weight enzyme 'is expressed as its ability to phosphorylate histone. The protein kinase activity of the pellet is increased between 3 and 17 hr after reserpine injection. Our evidence indicates that this increase is due to a translocation from cytosol to subcellular structures of a kinase that utilizes lysine-rich histone as phosphate acceptor. The protein kinase activity that is extracted from a purified nuclear fraction prepared from the adrenal medulla of rats injected 7 hr previously with reserpine is greater than that extracted from medulla of saline-treated rats.In neural tissue, intracellular communication is mediated by hormones and neurotransmitters. The latter not only regulate the polarity of the neuronal membranes, but they also influence the expression of the metabolic code in postsynaptic cells (1). In the rat adrenal medulla, the biosynthesis of tyrosine-3-monooxygenase [EC 1.14.16.2; L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating)] is regulated transsynaptically (2)(3)(4). In chromaffin cells that are presistently (1 hr or more) stimulated by the acetylcholine released by nerve impulses from presynaptic terminals, the monooxygenase is induced 18-24 hr later (2-7). We have adopted this transsynaptic induction of tyrosine monooxygenase as a model to study the molecular mechanisms whereby the release of acetylcholine regulates the expression of the metabolic code in postsynaptic cells. The first event in this induction is an increase in the 3':5'-cyclic AMP (cAMP) content, presumably by the activation of adenylate cyclase (4,7,8). When the synthesis rate of cAMP is enhanced, the number of cytosol protein kinase (EC 2.7.1.37; ATP:protein phosphotransferase) molecules that are activated is also increased (3). In the present paper, we report that in the adrenal medulla the transsynaptic regulation of the expression of the metabolic code is critically dependent on the translocation from cytosol into the nucleus of a low-molecular-weight catalytic subunit of protein kinase. Moreover, we show that this translocation, which begins a few hours after the increase in the release of acetylcholine elicited by reserpine, Abbreviation: cAMP, 3':5'-cyclic...

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Section: Methodsmentioning

confidence: 99%

Activation and nuclear translocation of protein kinase during transsynaptic induction of tyrosine 3-monooxygenase.

Costa¹,

Kurosawa²,

Guidotti³

1976

Proc. Natl. Acad. Sci. U.S.A.

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“…Previous studies with Ganaraska rainbow trout have shown that RNA concentration is a more sensitive indicator of recent growth (measured by changes in body size) than RNA:DNA ratio (Ferguson & Danzmann, 1990). The concentrations of RNA and DNA were measured in white muscle according to the flourometric method of Karsten & Wollenberger (1972) as modified by Ferguson & Drahushchak (1989).…”

Section: Electrophoresismentioning

confidence: 99%

Enzyme heterozygosity and growth in rainbow trout: genetic and physiological explanations

Ferguson

1992

Heredity

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The possibility of whether the association between enzyme heterozygosity and body size (fork length) is consistent among rainbow trout ( Oncorhynchus mykiss) with different degrees of relationship and is affected by fish age was determined. Six-month-old full-sibs and progeny groups from five and 13 parents of each sex do not show the positive associations between multilocus heterozygosity and fork length which are detectable in larger pooled gamete matings (259 x 25 ci). Moreover, the relationship between heterozygosity at single loci and fork length is inconsistent among families. These data suggest that chromosomal segments, marked by the enzyme loci, are responsible for the phenotypic effects (associative overdominance). Fish age affects both the strength and direction of the association between multilocus heterozygosity and fork length; the association is positive in 6-month-old rainbow trout but is weaker, negative, or differs between the sexes in fish at 1 year. Moreover, the strength of the relationship changes over time in repeated measurements on the same fish. A decline in growth of larger and more heterozygous fish, because of precocial sexual maturation, may partially explain the changing relationship between multilocus heterozygosity and fork length. Sexually mature males are significantly more heterozygous than immature males and show significantly reduced recent growth rates as measured by white muscle RNA concentration. However, females, who rarely mature at 1 year, show significantly higher recent growth rates. Thus, heterozygosity is differentially associated with the allocation of energy resources into somatic and reproductive tissue in male and female rainbow trout.

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“…The final preparation contained 2 mg calf thymus ssDNA/ml. DNA was determined by the double-wavelength method of Spirin [ll] as well as by a fluorescence method using ethidium bromide (to characterize the yield of cell nuclei [12]). The proteins were determined by the modified Lowry method, with crystalline albumin as a standard [13], and at 280 nm.…”

Section: Reagents and Determinationsmentioning

confidence: 99%

Exodeoxyribonuclease from rat brain specific for single‐stranded DNA

Иванов¹,

Gaziev²,

Tretyak³

1983

European Journal of Biochemistry

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An exodeoxynuclease with single-stranded DNA specificity has been isolated from rat brain and purified to electrophoretic homogeneity. Approximately 1 100-fold purification with a yield of 16 % has been achieved by chromatography on DNA-agarose, hydroxyapatite and Sephadex G-200. The enzyme has a pH optimum at 8.4, requires Mgz+ or MnZf as a cofactor and has a molecular weight of about 60000. It hydrolyzes homologous, heterologous, synthetic and depurinated substrates at the same rate liberating nucleoside 5'-monophosphates but does not attack ultraviolet-irradiated polydeoxyribonucleotides. This DNase is localized predominantly in neuronal cell nuclei and appears to be lacking in rat liver tissue.

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Determination of DNA and RNA in homogenized cells and tissues by surface fluorometry

Cited by 171 publications

References 20 publications

Activation and nuclear translocation of protein kinase during transsynaptic induction of tyrosine 3-monooxygenase.

Activation and nuclear translocation of protein kinase during transsynaptic induction of tyrosine 3-monooxygenase.

Enzyme heterozygosity and growth in rainbow trout: genetic and physiological explanations

Exodeoxyribonuclease from rat brain specific for single‐stranded DNA

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