The effect of Triton X-100 on the integrity of nuclear RNA isolated from rat cerebral cortex

The effect of convulsions, induced by flurothyl, on RNA synthesis in purified unfractionated nuclei and the cytoplasm of rat cerebral cortex was studied by using a double-label technique involving injection of [3H]- and [14C]-orotate intracisternally. 2. Intact RNA was extracted in 80% yield by an enzymic method by using a proteinase in the presence of sodium dodecyl sulphate followed by deoxyribonuclease. Electrophoresis on 1.5% polyacrylamide-0.5% agarose gels revealed the presence of giant nuclear RNA of size up to approx. 300 × 10(6) daltons and mRNA of maximal mol.wt. 9 × 10(6)-16 × 10(6). 3. Nuclear RNA synthesis was decreased to 27% in the first 15 min after convulsions but rapidly increased, so that at 1 1/2 h it was 124% of the control, and at 6 h 147%. 4. Labelling of cytoplasmic RNA was decreased to 15% at 15 min after convulsions but had not recovered to control values by 6 h. 5. Analysis of radioactive gel patterns and the 3H/14C ratio at six time-points (15 min-6h) showed that the major effect was inhibition of the processing of heterogeneous nuclear RNA resulting in a sharp decline in the export of newly synthesized RNA from the nucleus. 6. Cytoplasmic RNA patterns indicated that specific messengers were synthesized at different times during the recovery of the cell after convulsions.

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Stimulation of benzodiazepine receptors in native and triton x-100-treated membranes of rat cerebral cortex by GABA and its analogs

Brain Research Bulletin ◽

10.1016/0361-9230(79)90267-3 ◽

1979 ◽

Vol 4 (5) ◽

pp. 710

Author(s):

David T. Wong ◽

Frank P. Bymaster ◽

Penny T. Lane

Keyword(s):

Cerebral Cortex ◽

Benzodiazepine Receptors ◽

Rat Cerebral Cortex ◽

Triton X ◽

Stimulation Of

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The diglyceride kinase of rat cerebral cortex

Biochemical Journal ◽

10.1042/bj1220171 ◽

1971 ◽

Vol 122 (2) ◽

pp. 171-179 ◽

Cited By ~ 100

Author(s):

E. G. Lapetina ◽

J. N. Hawthorne

Keyword(s):

Cerebral Cortex ◽

Phosphatidic Acid ◽

Nerve Ending ◽

Osmotic Shock ◽

Sodium Deoxycholate ◽

Acid Synthesis ◽

Optimum Concentration ◽

Subcellular Fractions ◽

Rat Cerebral Cortex ◽

Triton X

1. Formation of phosphatidic acid by diglyceride kinase (EC 2.7.1.-) in the presence of ATP and Mg2+ was shown in a homogenate and subcellular fractions of rat cerebral cortex. 2. The kinase was activated by Mg2+. Ca2+ activated to a smaller extent but was inhibitory in the presence of optimum concentration of Mg2+. Activity was greatly increased in the presence of added 1,2-diglyceride. 3. Sodium deoxycholate markedly stimulated the reaction, but other detergents (Cutscum and Triton X-100) did not. 4. Diglyceride kinase was concentrated in the supernatant and microsomal fractions from rat cerebral cortex. The distribution of the kinase in the particulate fractions resembled that of acetylcholinesterase and 5′-nucleotidase. 5. The rate of phosphatidic acid synthesis by the diglyceride kinase route was much greater than reported rates for acylation of 3-glycerophosphate and was also very rapid in comparison with the rates of other steps in the synthesis of phosphoinositides. 6. Acetylcholine had no stimulatory effect on diglyceride kinase of isolated intact nerve-ending particles or of nerve-ending membranes obtained after osmotic shock.

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Glutamate decar☐ylase solubilized from the rat cerebral cortex by two different concentrations of triton x-100: Effects of glutamate analogues and analysis by SDS-page/western blotting using GAD6 and K2 antibodies

Neurochemistry International ◽

10.1016/0197-0186(94)00111-7 ◽

1995 ◽

Vol 26 (2) ◽

pp. 179-185 ◽

Cited By ~ 1

Author(s):

Sven O. Johanson ◽

Yi li ◽

Vladimir J. Balcar

Keyword(s):

Cerebral Cortex ◽

Western Blotting ◽

Rat Cerebral Cortex ◽

Sds Page ◽

Glutamate Analogues ◽

Triton X

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Rat cerebral-cortex adenosine deaminase activity and its subcellular distribution

Biochemical Journal ◽

10.1042/bj1440037 ◽

1974 ◽

Vol 144 (1) ◽

pp. 37-41 ◽

Cited By ~ 36

Author(s):

Ian Pull ◽

Henry McIlwain

Keyword(s):

Cerebral Cortex ◽

Adenosine Deaminase ◽

Subcellular Distribution ◽

Rat Cerebral Cortex ◽

Adenosine Deaminase Activity ◽

Triton X

A radioisotopic assay for adenosine deaminase (EC 3.5.4.4) is described together with its application in investigating the activity of the enzyme in rat cerebral cortex. Activity of the adenosine deaminase was determined to be 115nmol/min per g of tissue, measured in isoosmotic sucrose dispersions of the neocortex, and to be 170nmol/min per g of tissue after treatment with Triton X-100. The enzyme was concluded to be largely cytoplasmic, with a Km of 54–57μm for adenosine. Action of the deaminase, and other aspects of the metabolism of adenosine in intact neocortical tissue, were quantitatively appraised on the basis of the newly determined characteristics.

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