The uptake of 35S into rat tissues after injection of [35S]methionine

METHIONINE metabolism in oil;o, and its uptake into different organs, has received attention by a number of authors (GAITONDE and RICHTER, 1955, 1956) and the rnetabolic activity of this substance has been demonstrated (NIKLAS et a/., CLOUET and RICHTER, 1959). No attention has been paid, however, to methionine sulphoxide and methionine sulphone, the first and the second oxidation stages of methionine. The destribution of r5S]methionine sulphoxirnine in the rat has been investigated by ROTH et al. (1952) who in a later paper ( ROTH et al., 1953) suggested that the cause of the toxic effect of methionine sulphoxirnine could be the incorporation of this substance in place of methionine into the proteins. In connexion with our studies of the effect of methionine and methionine sulphoximine on epileptic fits in experimental animals ( KOLOUSEK, 1956,1958; KOLOUSEK and LODIN, 1959) we set ourselves the task of ascertaining whether or not methionine sulphoximine, and the sulphoxide and sulphone are incorporated into.proteins. M E T H O DAdult male white rats of 25C280 g were used. The [55S]-~~-methionine used was of Soviet origin and its specific activity was 800 mc/g. [35S]-~~-Methionine sulphoxide and its normer and [%]-DLmethionine sulphoximine and its normer were, in essence, prepared according to BENTLEY, MCDER- MOTT and WHITEHEAD (1951).[S5S]-~~-Methionine sulphone and its normer were prepared from [S5S]-~~-methionine by oxidation with performic acid and purification of the products by cation exchange.Methionine sulphoxide, methionine sulphone and methionine sulphoximine were injected intraperitoneally (400 mg/kg body weight), the substances containing "S being mixed with their normers so that the activity of the injected substances was about 1 mc/kg body weight. The dose of 400 mg/kg body weight was chosen because it corresponds to the paroxysmal dose of methionine sulphoximine which provokes in the rat a series of epileptic fits, the so-called sfatus epilepficus, 5-6 hr after i.p. application. Although the incorporation of [s5S]-methionine into brain has been studied before by other authors, we also applied [35S]-~~-methionine to the experimental rats (I mc/kg body weight, i.p.) for comparison with the above mentioned substances. All the applied preparations were chromatographically pure. The relatively low specific activity of the applied substances and the 24 hr duration of the experiment exclude any possibility of artefacts due to radioactive disintegration.After treatment, the experimental animals were kept in metabolic cages, according to COMMAR. receiving water ad libitum and the urine was collected in calibrated test-tubes. After 2. 6 and 24 hr the animals were killed by decapitation, the heads falling into liquid nitrogen. For each time-interval and for each substance 12 rats were used. The results are given as mean values. The frozen brain was taken from each head and ground to powder under liquid nitrogen. Samples of approximately I g in weight were further homogenized in glass homogenization test...

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“…1) suggesting lively metabolic activity of the brain. These results are similar to previous findings (GAITONDE and RICHTER, 1955).…”

Section: R E S U L T Ssupporting

confidence: 93%

“…The sulphur was oxidized to SO, and converted to benzidine sulphate which was quantitatively filtered off. The method used was essentially that of GAITONDE and RICHTER (1955). The precipitate, a disc 16 mm in diameter, was measured by a G-M tube-with a mica window of thickness 3.5 mg/cm*.…”

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confidence: 99%

THE METABOLISM OF METHIONINE DERIVATIVES CONTAINING ³⁵S IN THE RAT BRAIN IN VIVO

Kolousek

Babický

1961

Journal of Neurochemistry

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“…Twenty-four hours later the amount of the incorporated amino acid is decreased by about 20 per cent. GAITONDE and RICHTER (1955) found relatively little, if any, change within the first 24 hr. In our experiments the 20 per cent loss of 35S content of the brain in 24 hr was a constant finding.…”

Section: Resultsmentioning

confidence: 84%

QUANTITATIVE DETERMINATION OF THE UPTAKE OF [³⁵S]METHIONINE IN DIFFERENT REGIONS OF THE NORMAL RAT BRAIN

Merei¹,

Gallyas²

1964

Journal of Neurochemistry

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THE RATE of uptake of [35S]methionine in rat tissues has been extensively studied in the past ten years. FRIEDBERG, TARVER and GREENBERG (1948) found that after intraperitoneal or intravenous administration relatively large amounts of 35S appeared in the kidney, liver and other organs, while its concentration in brain remained comparatively low. Almost the same result was obtained by GAITONDE and RICHTER (1955, 1956), who developed a method for the isolation of sulphur after administration of r5S]methionine in vivo. They showed that the radioactivity of the examined tissue was due to [35S]methionine and a small amount of [35S]cysteine. On the basis of their experiments with rats the mean rate of methionine incorporation during the first 20-60 min after intracisternal injection was 0-4 ,uequiv./g proteinlhr. Their experiments also confirmed the findings of RUTMAN, DEMPSTER and TARVER (1949) who had reported that, although the rate of uptake of 35S by the brain was dependent on constitutional factors, consistent results could be obtained in animals of the same strain.The investigations of GAITONDE and RICHTER (1955, 1956) showed that measurement of the incorporation of r5S]methionine by the brain proteins was a convenient method for studying the protein metabolism of the CNS. Therefore, it seemed t o be worth while making a systematic quantitative study of the incorporation of [35S]methionine by the brain proteins in different regions of the normal CNS using the method already described by us (M~REI and GALLYAS, 1964). M E T H O D SNinety-seven albino rats of a home bred strain were divided into five groups: twenty-one were used to determine the rate of incorporation in the first 24 hr; twenty-four were used to measure the incorporation from 0-16 days after intraperitoneal administration of [S5S]methionine; twelve were used to study the effect of changes in the specific activity of the labelled amino acid upon the rate of incorporation; thirteen served to determine the pool of methionine in the whole body of the animal and twenty-seven were studied for the effect of starvation on the protein metabolism of the CNS.After intraperitoneal injection of 150 pc of [g5S]methionine (specific activity: 652 mc/g), three animals were killed at the times indicated in Figs. 1-3 to determine the rate of incorporation. The brain, the upper part of the spinal cord and the hypophysis were removed, fixed in acetone, embedded in paraffin and sectioned at 10 p. Embedding was accelerated by the use of vacuum throughout the procedure. After deparaffinization in chloroform the sections were covered with a 0.5 % solution of acrylic resin. Autoradiography and photometric evaluation were carried out as described in the previous paper.Histochemical treatment of sections with ribonuclease, hyaluronidase, trypsin and pepsin was carried out as described by PEARSE (1961).

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“…

THERE is abundant evidence that protein synthesis in mammalian cells as well as in microorganisms occurs predominantly on polyribosomes. However, in spite of numerous publications emphasizing the active synthesis of proteins in brain in vivo ( GAITONDE and RICHTER, 1955;WAELSCH and LAJTHA, 1961 ; ROBERTS and MORELOS, 1965) and in acellular systems (SATAKE, MASE, TAKAHASHI and OGATA, 1960; Acs, NEIDLE and WAELSCH, 1961 ; ZOMZELY, ROBERTS and RAPAPORT, 1964;SUZUKI, KOREY and TERRY, 1964; MURTHY and RAPPOPORT 1965a,b) only few attempts have been made to characterize polysomes in nervous tissue. HERRIMAN and HUNTER (1965) working with mouse brain and CAMPBELL, MAHLER, MOORE and TEWARI (1966) working with rat brain have shown the occurrence of particles heavier than monomeric ribosomes with ability to incorporate amino acids.

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confidence: 99%

Polysomes and Polysomal Rna of Rat Brain

Jacob

Samec

Stévenin

et al. 1967

Journal of Neurochemistry

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The uptake of 35S into rat tissues after injection of [35S]methionine

Cited by 78 publications

References 6 publications

THE METABOLISM OF METHIONINE DERIVATIVES CONTAINING ³⁵S IN THE RAT BRAIN IN VIVO

THE METABOLISM OF METHIONINE DERIVATIVES CONTAINING ³⁵S IN THE RAT BRAIN IN VIVO

QUANTITATIVE DETERMINATION OF THE UPTAKE OF [³⁵S]METHIONINE IN DIFFERENT REGIONS OF THE NORMAL RAT BRAIN

Polysomes and Polysomal Rna of Rat Brain

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The uptake of 35S into rat tissues after injection of [35S]methionine

Cited by 78 publications

References 6 publications

THE METABOLISM OF METHIONINE DERIVATIVES CONTAINING 35S IN THE RAT BRAIN IN VIVO

THE METABOLISM OF METHIONINE DERIVATIVES CONTAINING 35S IN THE RAT BRAIN IN VIVO

QUANTITATIVE DETERMINATION OF THE UPTAKE OF [35S]METHIONINE IN DIFFERENT REGIONS OF THE NORMAL RAT BRAIN

Polysomes and Polysomal Rna of Rat Brain

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THE METABOLISM OF METHIONINE DERIVATIVES CONTAINING ³⁵S IN THE RAT BRAIN IN VIVO

THE METABOLISM OF METHIONINE DERIVATIVES CONTAINING ³⁵S IN THE RAT BRAIN IN VIVO

QUANTITATIVE DETERMINATION OF THE UPTAKE OF [³⁵S]METHIONINE IN DIFFERENT REGIONS OF THE NORMAL RAT BRAIN