BREAKDOWN OF [3H]OXYTOCIN BY RAT TISSUES IN VITRO

1973 ◽  
Vol 59 (2) ◽  
pp. 305-312
Author(s):  
SANDRA M. EGAN ◽  
A. LIVINGSTON
Keyword(s):  
Skeletal Muscle ◽  
Mammary Gland ◽  
Incubation Medium ◽  
Rat Tissues ◽  
Tissue Uptake ◽  
Tissue Extracts ◽  
Rat Mammary Gland

SUMMARY Tissue uptake of oxytocin by rat mammary gland, uterus, heart and skeletal muscle was demonstrated by incubation of these tissues with 3H-labelled oxytocin. Chromatography of tissue extracts showed that some breakdown of oxytocin had occurred after only 30 s. However, in all cases there was no breakdown of oxytocin in the corresponding incubation medium. This indicated that the breakdown of oxytocin occurred within the tissues.

scholarly journals Expression level of Ubc9 protein in rat tissues.

2003 ◽  
Vol 50 (4) ◽  
pp. 1065-1073 ◽  
Author(s):  
Filip Gołebiowski ◽  
Aneta Szulc ◽  
Monika Sakowicz ◽  
Andrzej Szutowicz ◽  
Tadeusz Pawełczyk
Keyword(s):  
Skeletal Muscle ◽  
Mrna Level ◽  
Protein Band ◽  
Expression Level ◽  
Rat Tissues ◽  
Mrna Levels ◽  
Tissue Extracts ◽  
Ubiquitin Conjugating Enzyme ◽  
High Level

Ubc9 is a homologue of the E2 ubiquitin conjugating enzyme and participates in the covalent linking of SUMO-1 molecule to the target protein. In this report we describe a simple and efficient method for obtaining pure human recombinant Ubc9 protein. The purified Ubc9 retained its native structure and was fully active in an in vitro sumoylation assay with the promyelocytic leukaemia (PML) peptide as a substrate. In order to better understand the physiology of Ubc9 protein we examined its levels in several rat tissues. Immunoblot analyses performed on tissue extracts revealed quantitative and qualitative differences in the expression pattern of Ubc9. The Ubc9 protein was present at a high level in spleen and lung. Moderate level of Ubc9 was detected in kidney and liver. Low amount of Ubc9 was observed in brain, whereas the 18 kDa band of Ubc9 was barely visible or absent in heart and skeletal muscle. In heart and muscle extracts the Ubc9 antibodies recognized a 38 kDa protein band. This band was not visible in extracts of other rat tissues. A comparison of the relative levels of Ubc9 mRNA and protein indicated that the overall expression level of Ubc9 was the highest in spleen and lung. In spleen, lung, kidney, brain, liver and heart there was a good correlation between the 18 kDa protein and Ubc9 mRNA levels. In skeletal muscle the Ubc9 mRNA level was unproportionally high comparing to the level of the 18 kDa protein. The presented data indicate that in the rat the expression of the Ubc9 protein appears to have some degree of tissue specificity.

scholarly journals Identification in skeletal muscle of a distinct extracellular pool of amino acids, and its role in protein synthesis

1971 ◽  
Vol 121 (5) ◽  
pp. 817-827 ◽  
Author(s):  
R. C. Hider ◽  
E. B. Fern ◽  
D. R. London
Keyword(s):  
Skeletal Muscle ◽  
Amino Acids ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Incubation Medium ◽  
Extensor Digitorum Longus ◽  
Extensor Digitorum ◽  
Longus Muscle ◽  
Kinetics Of

1. The kinetics of radioactive labelling of extra- and intra-cellular amino acid pools and protein of the extensor digitorum longus muscle were studied after incubations with radioactive amino acids in vitro. 2. The results indicated that an extracellular pool could be defined, the contents of which were different from those of the incubation medium. 3. It was concluded that amino acids from the extracellular pool, as defined in this study, were incorporated directly into protein.

Properties of 5′-deiodinase of 3,3′,5′-triiodothyronine in rat skeletal muscle

1989 ◽  
Vol 120 (1) ◽  
pp. 69-74 ◽  
Author(s):  
Fujiko Tsukahara ◽  
Teruko Nomoto ◽  
Michiko Maeda
Keyword(s):  
Skeletal Muscle ◽  
Drinking Water ◽  
Incubation Medium ◽  
Marked Reduction ◽  
Daily Administration ◽  
Type I ◽  
Rat Skeletal Muscle ◽  
Thyroid Status ◽  
Altered Thyroid

Abstract. To characterize rT3 5′-deiodinase (5′D) in rat skeletal muscle, the effects of altered thyroid status and PTU on rT3 5′D were studied. rT3 5′D activity was measured by incubating homogenates of rat skeletal muscle with [125]rT3, iodine labelled in the outer ring, in the presence of 20 mmol/l DL-dithiothreitol. This activity was observed to increase significantly 24 h after a single sc injection of T3 (75 μg/kg). The increase following the daily administration of this drug (15 or 75 μg/kg) for 3 and 14 days was dependent on the dose and number of previous days of injection. A significant decrease in activity was observed 2 weeks after thyroidectomy. The addition of 0.1 mmol/l 6-n-propyl-2-thiouracil (PTU) to the incubation medium in vitro caused a marked reduction in the activity in homogenates of skeletal muscle from hypothyroid, euthyroid and hyperthyroid rats. PTU, present at 0.05% in the drinking water for 2 weeks virtually abolished it. The properties of rT3 5′D in rat skeletal muscle thus appear to be essentially the same as those of type I enzyme with respect to response toward altered thyroid status and PTU.

scholarly journals Phosphoribosyl pyrophosphate and phosphoribosyl pyrophosphate synthetase in rat mammary gland. Changes in the lactation cycle and effects of diabetes, insulin and phenazine methosulphate

1986 ◽  
Vol 238 (2) ◽  
pp. 553-559 ◽  
Author(s):  
S Kunjara ◽  
M Sochor ◽  
N Salih ◽  
P McLean ◽  
A L Greenbaum
Keyword(s):  
Mammary Gland ◽  
Fold Increase ◽  
Pentose Phosphate ◽  
Diabetic Rat ◽  
Synthetase Activity ◽  
Phosphoribosyl Pyrophosphate ◽  
Rat Mammary Gland ◽  
Lactation Cycle ◽  
Phenazine Methosulphate

Changes in the tissue content of phosphoribosyl pyrophosphate (PPRibP), glucose 6-phosphate, ribose 5-phosphate (Rib5P), RNA and DNA, of the activity of PPRibP synthetase (EC 2.7.6.1) and the conversion of [1-14C]- and [6-14C]-glucose into 14CO2 were measured at mid-lactation in the normal and diabetic rat and in pregnancy, lactation and mammary involution in the normal rat. The PPRibP, glucose 6-phosphate and Rib5P contents increase during pregnancy and early lactation to reach a plateau value at mid-lactation, before falling sharply during weaning. The PPRibP content, PPRibP synthetase activity and flux of glucose through the oxidative pentose phosphate pathway (PPP) all change in parallel during the lactation cycle. Similarly, after 3 and 5 days duration of streptozotocin-induced diabetes, ending on day 10 of lactation, there were parallel declines in PPRibP content, PPRibP synthetase and PPP activity. The effect of streptozotocin was prevented by pretreatment with nicotinamide and partially reversed by insulin administration. Addition of insulin to lactating rat mammary-gland slices incubated in vitro significantly raised the PPRibP content (+47%) and the activity of the PPP (+40%); phenazine methosulphate, which gives a 2-fold increase in PPP activity, raised the PPRibP content of lactating mammary gland slices by approx. 3-fold. It is concluded that Rib5P, generated in the oxidative segment of the PPP, is an important determinant of PPRibP synthesis in the lactating rat mammary gland and that insulin plays a central role in the regulation of the bioavailability of this precursor of nucleotide and nucleic acid synthesis.

THE IN VITRO METABOLISM OF MAMMARY GLAND SLICES IN THE PRESENCE OF POSTERIOR PITUITARY LOBE EXTRACTS RICH IN MELANOPHORE-DISPERSING ACTIVITY

1957 ◽  
Vol 15 (4) ◽  
pp. 366-373 ◽  
Author(s):  
T. R. BRADLEY ◽  
G. M. MITCHELL
Keyword(s):  
Mammary Gland ◽  
Guinea Pigs ◽  
Incubation Medium ◽  
Mammary Glands ◽  
Gas Evolution ◽  
Early Lactation ◽  
Posterior Pituitary ◽  
Rats And Mice ◽  
Posterior Pituitary Lobe

SUMMARY Slices cut from mammary glands of rats and mice during gestation and lactation were incubated in vitro in the presence of pig posterior pituitary lobe extracts rich in melanophore-dispersing ('B') activity. Slices taken in early lactation but not during gestation or late lactation showed increased net gas evolution compared with control slices. Similar tissue from rabbits and guinea-pigs did not give rise to this effect, nor did slices of other tissues taken from lactating rats. The increased net gas evolution was not observed in the absence of glucose from the incubation medium. Treatment of the 'B' extract with NaOH or hypophysectomy of the rats prior to use decreased the response.

scholarly journals HISTOCHEMICAL DEMONSTRATION OF PHOSPHOLIPASE B (LYSOLECITHINASE) ACTIVITY IN RAT TISSUES

1966 ◽  
Vol 14 (12) ◽  
pp. 907-914 ◽  
Author(s):  
ATHOS OTTOLENGHI ◽  
JOHN P. PICKETT ◽  
WILLIAM B. GREENE
Keyword(s):  
Fatty Acid ◽  
Incubation Medium ◽  
Copper Ions ◽  
Isotonic Saline ◽  
Histochemical Demonstration ◽  
Interstitial Tissue ◽  
Rat Tissues ◽  
Positive Staining ◽  
Phospholipase B

A method has been developed for the histochemical demonstration of phospholipase B (lysolecithinase) of rat tissues. The enzyme attacks lysolecithin with liberation of 1 mole of glycerylphosphorylcholine and 1 mole of fatty acid. The recommended procedure involves use of 6-10 µ frozen sections, fixed in cold calcium-formol and incubated at 37°C in Tris buffered medium at pH 6.6 containing 2.2 x 10–3 M lysolecithin and 1% cobalt acetate. The fatty acid liberated by enzymatic hydrolysis is trapped as a cobalt precipitate and is then converted to a blackbrown precipitate by treatment with dilute ammonium sulfide in cold isotonic saline. Equivalent amounts of fatty acid and glycerylphosphorylcholine are recovered by extraction and analysis of the incubated sections and of the incubation medium, thus proving that lysolecithin hydrolysis occurs under the proposed reaction conditions. Staining is reduced by treating the sections with copper ions, mercury compounds, alcohols, acetone and by heating at 60°C prior to incubation with substrate. Lowering of the pH of the incubation medium has similar effect. These findings are interpreted as evidence of the enzymatic nature of the reaction. Cells exhibiting a positive staining are found in the lamina propria of the intestinal villi and crypts, in the red pulp of the spleen and in the interstitial tissue of lung, liver and thymus. Similar elements are present in bone marrow smears and in leukocyte preparations obtained by peritoneal lavage. The morphologic and staining characteristics of these cells correspond to those of the eosinophilic leukocytes. Physical and chemical agents (x-irradiation corticosteroids) which sharply decrease the number of eosinophils also reduce the number of cells shown histochemically to hydrolyze lysolecithin. A correspondent. diminution of phospholipase B activity of homogenates of the same tissues can be shown in vitro. Differences in tissue distribution and chemical properties distinguish the phospholipase B from less specific esterases and lipases.

Release of enzymes from rat mammary gland, skeletal muscle, and liver

Life Sciences ◽  
1970 ◽  
Vol 9 (5) ◽  
pp. 291-300 ◽  
Author(s):  
Börje W. Karlsson ◽  
Enar I. Carlsson
Keyword(s):  
Skeletal Muscle ◽  
Mammary Gland ◽  
Rat Mammary Gland

Metabolism of n-hexane by rat liver and extrahepatic tissues and the effect of cytochrome P-450 inducers

1997 ◽  
Vol 16 (3) ◽  
pp. 131-137 ◽  
Author(s):  
Sarah J Crosbie ◽  
PG Blain ◽  
Faith M Williams
Keyword(s):  
Skeletal Muscle ◽  
Rat Liver ◽  
Fold Increase ◽  
Rat Tissues ◽  
Human Cytochrome ◽  
Extrahepatic Tissues ◽  
Mass Spectometry ◽  
Fast Twitch ◽  
Cytochrome P 450

1 The in vitro metabolism ofn-hexane was studied in rat liver, lung, brain and skeletal muscle microsomes and in microsomes prepared from cell lines expressing human cytochrome P-450 2E1 or 2B6. The hydro xylated metabolites ofn-hexane were quantified by gas chromatography-mass spectometry. 2 Rat liver and extensor digitorum longus (EDL, fast- twitch skeletal muscle) microsomes and the CYP 2B6 microsomes produced the pre-neurotoxic metabolite of n-hexane, 2-hexanol as a major metabolite in contrast to the other rat tissues examined. 3 Inhibition of 2- and 3-hexanol production from n- hexane by rat lung microsomes using metyrapone, an inhibitor of cytochrome P-450 2B1 activity, resulted in almost complete inhibition of lung microsomal activ ity. 4 Production of all three hexanols was significantly increased with phenobarbital-induced rat liver micro somes, with a 10-fold increase in 2- and 3-hexanol production. A slight increase in 2-hexanol production with phenobarbital-induced rat EDL and brain micro somes was observed. No increase in n-hexane meta bolism was noted following induction with β- naphthoflavone or with ethanol.

scholarly journals Rapid inhibition of lipogenesis in vivo in lactating rat mammary gland by medium- or long-chain triacylglycerols and partial reversal by insulin

1980 ◽  
Vol 192 (1) ◽  
pp. 361-364 ◽  
Author(s):  
L Agius ◽  
D H Williamson
Keyword(s):  
Mammary Gland ◽  
Glucose Utilization ◽  
Insulin Administration ◽  
Hepatic Lipogenesis ◽  
Medium Chain ◽  
Rat Mammary Gland ◽  
Long Chain ◽  
Partial Reversal

An intragastric load of medium- or long-chain triacylglycerols inhibited lipogenesis in lactating rat mammary gland in vivo by 82 or 89% respectively. This inhibition was reversed partially by insulin administration. Long-chain triacylglycerols inhibited hepatic lipogenesis in vivo but medium-chain triacylglycerols increased it 2-fold. Glucose utilization in vitro by mammary gland acini from triacylglycerol-fed rat was normal.

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