scholarly journals Role of haem in the induction of cytochrome P-450 by phenobarbitone. Studies in chick embryos in ovo and in cultured chick embryo hepatocytes

1981 ◽  
Vol 198 (2) ◽  
pp. 321-329 ◽  
Author(s):  
U Giger ◽  
U A Meyer
Keyword(s):  
Chick Embryo ◽  
Inhibitory Effect ◽  
Chick Embryos ◽  
In Ovo ◽  
Haem Biosynthesis ◽  
Rate Limiting ◽  
Cytochrome P 450 ◽  
Hepatic Cytochrome

The role of haem synthesis during induction of hepatic cytochrome P-450 haemoproteins was studied in chick embryo in ovo and in chick embryos hepatocytes cultured under chemically defined conditions. 1. Phenobarbitone caused a prompt increase in the activity of 5-aminolaevulinate synthase, the rate-limiting enzyme of haem biosynthesis, and in the concentration of cytochrome P-450. This induction response occurred without measurable initial destruction of the haem moiety of cytochrome P-450. 2. When intracellular haem availability was enhanced by exogenous haem or 5-aminolaevulinate, phenobarbitone-medicated induction of cytochrome P-450 was not affected in spite of the well known repression of 5-aminolaevulinate synthase by haem. These data are consistent with the concept that haem does not regulate the synthesis of cytochrome P-450 haemoproteins. 3. Acetate inhibited haem biosynthesis at the level of 5-aminolaevulinate formation. When intracellular haem availability was diminished by treatment with acetate, phenobarbitone-medicated induction was decreased. 4. This inhibitory effect of acetate on cytochrome P-450 induction was reversed by exogenous haem or its precursor 5-aminolaevulinate. These data suggest that inhibition of haem biosynthesis does not decrease synthesis of apo-cytochrome P-450. Moreover, they indicate that exogenous haem can be incorporated into newly formed aop-cytochrome P-450.

scholarly journals Decrease in hepatic cytochrome P-450 by cobalt. Evidence for a role of cobalt protoporphyrin

1982 ◽  
Vol 204 (1) ◽  
pp. 103-109 ◽  
Author(s):  
J F Sinclair ◽  
P R Sinclair ◽  
J F Healey ◽  
E L Smith ◽  
H L Bonkowsky
Keyword(s):  
Chick Embryo ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Cobalt Protoporphyrin ◽  
Cytochrome P 450 ◽  
Hepatic Cytochrome

Exposure of cultured chick-embryo hepatocytes to increasing concentrations of CoCl2 in the presence of allylisopropylacetamide results in formation of cobalt protoporphyrin, with a reciprocal decrease in haem and cytochrome P-450. Treatment of rats with CoCl2 (84 mumol/kg) and 5-aminolaevulinate (0.2 mmol/kg) also results in formation of cobalt protoporphyrin and a decrease in cytochrome P-450 in the liver. Hepatic microsomal fractions from rats treated with phenobarbital, CoCl2 and 5-aminolaevulinate were analysed by polyacrylamide gel electrophoresis. Cobalt protoporphyrin was associated mainly with proteins of 50000-53000 mol.wt. The results suggest that the formation of cobalt protoporphyrin occurred at the expense of the synthesis of haem, leading to a decrease in cytochrome P-450. Furthermore, the cobalt protoporphyrin that was formed may itself have been incorporated into apocytochrome P-450.

scholarly journals Drug-induced accumulation of uroporphyrin in chicken hepatocyte cultures. Structural requirements for the effect and role of exogenous iron

1984 ◽  
Vol 224 (3) ◽  
pp. 769-777 ◽  
Author(s):  
A Ferioli ◽  
C Harvey ◽  
F De Matteis
Keyword(s):  
Chick Embryo ◽  
Molecular Target ◽  
Chick Embryos ◽  
In Ovo ◽  
Drug Induced ◽  
Structural Requirements ◽  
Binding Characteristics ◽  
Alternative Hypotheses ◽  
Do So

The ability of drugs to cause uroporphyria in hepatocytes from 17-day-old chick embryos has been investigated and the response of the cells in culture compared with that of the intact liver of the embryos in ovo. In this chick-embryo system, drugs that cause accumulation of uroporphyrin within 19-24 h can only do so in culture; in contrast, 2-allyl-2-isopropylacetamide and 3,5-diethoxycarbonyl-1,4-dihydrocollidine, which stimulate production of protoporphyrin, are effective both in culture and in ovo. A role of exogenous iron in worsening drug-induced uroporphyria was demonstrated in cultures of hepatocytes; iron also caused preferential accumulation of uroporphyrin from added 5-aminolaevulinate in the absence of a porphyrogenic chemical. Uroporphyria was induced in cultures of hepatocytes by drugs of widely different structures, suggesting that the primary molecular target with which they interact may be relatively aspecific in its binding characteristics. These results are briefly discussed, and two alternative hypotheses for the drug-induced effect in uroporphyrinogen metabolism are considered.

scholarly journals Inhibition of uroporphyrinogen decarboxylase activity. The role of cytochrome P-450-mediated uroporphyrinogen oxidation

1990 ◽  
Vol 269 (2) ◽  
pp. 437-441 ◽  
Author(s):  
R W Lambrecht ◽  
J M Jacobs ◽  
P R Sinclair ◽  
J F Sinclair
Keyword(s):  
Chick Embryo ◽  
Fold Increase ◽  
Treated Mouse ◽  
Decarboxylase Activity ◽  
Chick Embryos ◽  
Uroporphyrinogen Decarboxylase ◽  
Chemically Induced ◽  
Cytochrome P 450 ◽  
Generating System

It was previously shown that uroporphyrinogen oxidation is catalysed by a form of cytochrome P-450 induced by 3-methylcholanthrene [Sinclair, Lambrecht & Sinclair (1987) Biochem. Biophys. Res. Commun. 146, 1324-1329]. We have now measured uroporphyrinogen oxidation and uroporphyrinogen decarboxylation simultaneously in 10,000 g supernatants from the livers of methylcholanthrene-treated mice and chick embryos incubated with an NADPH-generating system. We found that uroporphyrinogen oxidation is associated with inhibition of uroporphyrinogen decarboxylase activity. The decreased uroporphyrinogen decarboxylase activity was not due to depletion of substrate, since decarboxylase activity was not increased by a 2.6-fold increase in uroporphyrinogen. Uroporphyrinogen oxidation and the associated inhibition of decarboxylase activity were also observed with liver supernatant from methylcholanthrene-treated chick embryo; both actions required the addition of 3,3′,4,4′-tetrachlorobiphenyl. Uroporphyrinogen oxidation catalysed by microsomes from a methylcholanthrene-treated mouse inhibited the uroporphyrinogen decarboxylase activity in the 100,000 g supernatant. Ketoconazole, an inhibitor of cytochrome P-450, prevented both uroporphyrinogen oxidation and the inhibition of uroporphyrinogen decarboxylation. The addition of ketoconazole to mouse supernatant actively oxidizing uroporphyrinogen inhibited the oxidation and restored decarboxylation. The latter finding suggested that a labile inhibitor was formed during the oxidation. These results suggest uroporphyrinogen oxidation may be important in the mechanism of chemically induced uroporphyria.

scholarly journals Evidence that in chick embryos destruction of hepatic microsomal cytochrome P-450 haem is a general mechanism of induction of δ-aminolaevulinate synthase by porphyria-causing drugs

1980 ◽  
Vol 190 (3) ◽  
pp. 519-526 ◽  
Author(s):  
L K Lim ◽  
G Srivastava ◽  
J D Brooker ◽  
B K May ◽  
W H Elliott
Keyword(s):  
Chick Embryo ◽  
Enzyme Induction ◽  
General Mechanism ◽  
Chick Embryos ◽  
In Ovo ◽  
Microsomal Cytochrome ◽  
Primary Mechanism ◽  
Embryo Liver ◽  
The Relationship ◽  
Cytochrome P 450

A variety of prophyrinogenic compounds were tested for their effect in ovo on chick-embryo liver microsomal cytochrome P-450 haem concentration and mitochondrial delta-aminolaevulinate synthase activity. With all drugs tested, there was a 30—50% decrease in cytochrome P-450 haem concentration within 1 h of treatment, and this was closely followed by an increase in delta-aminolaevulinate synthase activity. The relationship was independent of the extent of enzyme induction and is consistent with the proposal that drug-mediated destruction of cytochrome P-450 haem is the primary mechanism of induction of delta-aminolaevulinate synthase. After induction, synthesis of delta-aminolaevulinate synthase could be maintained by inhibiting further haem synthesis. These studies suggest that induction of porphyria is a combination of two distinct processes: (a) induction of delta-aminolaevulinate synthase synthesis by destruction of cytochrome P-450 haem and consequent depletion of cellular free haem; (b) maintenance of continued delta-aminolaevulinate synthase synthesis by preventing replenishment of cellular haem either by inhibiting haem synthesis and/or by promoting continuous removal of newly synthesized haem.

The role of movement in the development of joints and related structures: the head and neck in the chick embryo

Development ◽  
1969 ◽  
Vol 22 (3) ◽  
pp. 349-371
Author(s):  
P. D. F. Murray ◽  
Daniel B. Drachman
Keyword(s):  
Skeletal Muscle ◽  
Chick Embryo ◽  
Normal Development ◽  
Neuromuscular Blocking ◽  
Neuromuscular Blocking Agents ◽  
Chick Embryos ◽  
In Ovo ◽  
Joint Development ◽  
Embryonic Life

Skeletal muscle contractions are necessary during embryonic life for the normal development of joints. The general features of joint development in immobile limbs were first studied with the techniques of grafting and organ culture. (Murray, 1926; Murray & Selby, 1930; Fell, 1925; Fell & Canti, 1934; Hamburger & Waugh, 1940; Lelkes, 1958). However, these methods of necessity entail a drastic alteration in the environment of the developing articulations, which may result in gross distortions of the skeletal structures themselves. More recently, neuromuscular blocking agents have been used to produce paralysis of chick embryos in ovo. When administered intravenously, these pharmacological substances produce profound paralysis, which may be continued for prolonged periods during embryonic development (Drachman & Coulombre, 1962a, b). Drachman & Sokoloff (1966) have analyzed the primary development of the knee, ankle and toe joints of the chick embryo by the use of these methods.

Essential role of the polyamines in early chick embryo development

Development ◽  
1980 ◽  
Vol 60 (1) ◽  
pp. 83-92
Author(s):  
B. Löwkvist ◽  
O. Heby ◽  
H. Emanuelsson
Keyword(s):  
Essential Role ◽  
Decisive Role ◽  
Chick Embryos ◽  
Irreversible Inhibitor ◽  
In Ovo ◽  
Polyamine Synthesis ◽  
Major Increase ◽  
Rate Limiting ◽  
Second Peak

The polyamines putrescine, spermidine and spermine were analyzed in chick embryos during the first 2 days of development. A rapid increase in the activity of ornithine decarboxylase (ODC), the initial and rate-limiting enzyme in polyamine synthesis, was observed immediately after onset of incubation. Peak activities were found at 15 and 23 h of incubation. The first peak coincides with gastrulation and the second peak with early neurulation in the embryo. All three polyamines varied in a similar manner as did ODC, with putrescine and spermidine at about the same level and spermine at a lower level. The ODC activity was blocked by α-difluoromethylornithine, an enzyme-activated irreversible inhibitor. The inhibitor was administered to embryos in ovo at 5 h of incubation, i.e. prior to the first major increase in ODC activity. This block prevented the accumulation of the polyamines and inhibited development at gastrulation, suggesting a decisive role for polyamines in this developmental event.

scholarly journals Haem synthesis from exogenous 5-aminolaevulinate in cultured chick-embryo hepatocytes. Effects of inducers of cytochromes P-450

1987 ◽  
Vol 248 (1) ◽  
pp. 229-236 ◽  
Author(s):  
S I Shedlofsky ◽  
P R Sinclair ◽  
H L Bonkovsky ◽  
J F Healey ◽  
A T Swim ◽  
...  
Keyword(s):  
Chick Embryo ◽  
Iron Chelator ◽  
Cellular Concentration ◽  
Haem Biosynthesis ◽  
Cytochrome P 450 ◽  
In Cells

The effects of inducers of cytochrome P-450 on haem biosynthesis from 5-aminolaevulinate were examined by using cultured chick-embryo hepatocytes. Cultures treated with either 2-propyl-2-isopropylacetamide or 3-methylcholanthrene contained increased amounts of cytochrome P-450 and haem. After treatment for 3 h with 5-amino[4-14C]laevulinate, the relative amounts of radioactivity accumulating as haem corresponded to the relative amounts of total cellular haem, but not to increases in the amounts of cytochrome P-450. Treatment with 5-aminolaevulinate did not alter cellular haem or cytochrome P-450 concentrations in either control or drug-treated cultures. The mechanism of the enhanced accumulation of radioactivity in haem was investigated. Although 2-propyl-2-isopropylacetamide enhanced the uptake of 5-aminolaevulinate and increased the cellular concentration of porphobilinogen 1.5-fold, these changes did not account for the increases in haem radioactivity. The inducing drugs had no effect on the rates of degradation of radioactive haem, but appeared to enhance conversion of protoporphyrin into haem. This latter effect was shown by: (1) a decreased accumulation of protoporphyrin from 5-aminolaevulinate in cells treated with inducers, and (2) complete prevention of this decrease if the iron chelator desferrioxamine was present. We conclude that inducers of cytochrome P-450 may increase haem synthesis not only by increasing activity of 5-aminolaevulinate synthase, but also by increasing conversion of protoporphyrin into haem.

scholarly journals Regulation of hepatic haem metabolism. Disparate mechanisms of induction of haem oxygenase by drugs and metals

1988 ◽  
Vol 250 (1) ◽  
pp. 189-196 ◽  
Author(s):  
B C Lincoln ◽  
J F Healey ◽  
H L Bonkovsky
Keyword(s):  
Chick Embryo ◽  
Primary Culture ◽  
In Ovo ◽  
Oxygenase Activity ◽  
Haem Oxygenase ◽  
Cytochrome P 450

We studied drug- and metal-mediated increases in activity of haem oxygenase, the rate-controlling enzyme for haem breakdown, in chick-embryo hepatocytes in ovo and in primary culture. Phenobarbitone and phenobarbitone-like drugs (glutethimide, mephenytoin), which are known to increase concentrations of an isoform of cytochrome P-450 in chick-embryo hepatocytes, were found to increase activities of haem oxygenase as well. In contrast, 20-methylcholanthrene, which increases the concentration of a different isoform of cytochrome P-450, had no effect on activity of haem oxygenase. Inhibitors of haem synthesis, 4,6-dioxoheptanoic acid or desferrioxamine, prevented drug-mediated induction of both cytochrome P-450 and haem oxygenase in embryo hepatocytes in ovo or in culture. Addition of haem restored induction of both enzymes. These results are interpreted to indicate that phenobarbitone and its congeners induce haem oxygenase by increasing hepatic haem formation. In contrast, increases in haem oxygenase activity by metals such as cobalt, cadmium and iron were not dependent on increased haem synthesis and were not inhibited by 4,6-dioxoheptanoic acid. We conclude that (1) induction of hepatic haem oxygenase activity by phenobarbitone-type drugs is due to increased haem formation, and (2) induction of haem oxygenase by drugs and metals occurs by different mechanisms.

Studies on regulatory mechanisms of heme biosynthesis in hepatocytes from normal and experimental-diabetic rats. Role of insulin

1990 ◽  
Vol 68 (6) ◽  
pp. 914-921 ◽  
Author(s):  
Eduardo T. Cánepa ◽  
Elena B. C. Llambías ◽  
Moisés Grinstein
Keyword(s):  
Aminolevulinic Acid ◽  
Rat Hepatocytes ◽  
Isolated Hepatocytes ◽  
Inhibitory Effect ◽  
Diabetic Rats ◽  
Significant Inhibition ◽  
Diabetic Rat ◽  
Experimental Conditions ◽  
Cytochrome P 450

In the present work we demonstrate that insulin decreases the phenobarbital-induced activities of δ-aminolevulinic acid synthase and ferrochelatase in isolated hepatocytes from normal and experimental-diabetic rats. Insulin concentrations required to produce significant inhibition in diabetic hepatocytes were higher than in normal cells. Under similar experimental conditions, insulin decreased the basal activities of δ-aminolevulinic acid synthase and ferrochelatase in hepatocytes from normal rats; no inhibitory effect was observed on the basal activity of δ-aminolevulinic acid synthase in hepatocytes from diabetic rats. Cytochrome P-450 content of both normal and diabetic cells was not affected by insulin in absence or presence of phenobarbital. The inhibitory action of insulin was exerted even when effective concentrations of glucagon, dexamethasone, or 8-(p-chlorophenylthio)-cAMP were present.Key words: δ-aminolevulinic acid synthase, ferrochelatase, cAMP, insulin, diabetic rat hepatocytes.

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