Liver Monoamine Oxidase Research Articles

Acute effects of organotins on brain, liver and kidney in rats.

Effects of dioctyltin oxide (DOTO) tricyclohexyltin hydroxide (TCHTOH) and tributyltin oxide (TBTO) were examined on some enzymic activities in liver and kidney and biogenic amines level in brain of rats at 24 hours after single subcutaneous administration (25 μmole/100 g B. Wt.). All the organotin compounds produced a significant increase in the activity of alkaline phosphatase and adenosin triphosphatase and decrease in monoamine oxidase in both liver and kidney. DOTO and TCHTOH were more effective in impairing the activity of succinate dehydro-genase in liver. Concentrations of γ-aminobutyric acid (GABA) and dopamine were found to be significantly decreased in brain however, acetylcholine concentration remained unaltered. These results suggest that organotin compounds DOTO and TCHTOH are more toxic to rats than TBTO.

P3B0110) - Electrophoretic properties of mitochondrial monoamine oxidase in monkey liver

Monkey liver monoamine oxidase (MAO) was predominantly the B-form enzyme from the observed differences in substrate specificities and differences in sensitivities to MAO inhibitors. It is known that a MAO inhibitor, pargyline, binds to MAO irreversibly in the molar ratio of 1:1. 3H-pargyline was used as a marker to determine the existence of MAO. The molecular weight of MAO in monkey liver mitochondria was investigated by SDS-polyacrylamide gel electrophoresis after solubilization of 3H-pargyline binding mitochondria with 6% sodium dodecyl sulphate (SDS). The subunit molecular weight was found to be 60,000. The molecular weight determined from the electrophoretic mobility on several concentrations of gels by disc gel electrophoresis in the absence of SDS was found to be 120,000. These results indicate that monkey liver mitochondrial MAO exists as a dimer. Isoelectric focusing of the enzyme after solubilization with 0.1% Triton X-100 and 0.75% Triton X-100 and 0.75% Lubrol showed that it had a pI value near 6.5. Similar pI values were obtained for enzyme preparations solubilized with 0.75% Triton X-100 after treatment with phospholipase A or methylethylketone. These results suggest that the pI value of MAO in monkey liver mitochondria does not depend on the properties of detergents used to solubilize the enzyme preparation.

Serotonin and the Control of Salivation in the Blowfly Calliphora

ABSTRACT The possibility that serotonin acts as a neurohormone stimulating salivation in the blowfly Calliphora vicina was studied by investigation of salivation induced by injections of high-potassium saline. Induced salivation is rapid and appears to be mediated by an active factor released into the haemolymph (High Potassium Salivary Gland Factor: HKSGF) since it is antagonized by cadmium (a calcium channel blocker) and by gramine (a serotonin-receptor blocker). The action of HKSGF on salivary glands in vitro is indistinguishable from that of serotonin: (a) it generates serotonin-like transepithelial potential changes, (b) its effect on salivation is antagonized by gramine, (c) it is as heat stable as serotonin, (d) it has the same solubility in a variety of organic solvents, (e) it is unaffected by incubation with leucine aminopeptidase or trypsin and (d) it is inactivated by rat liver monoamine oxidase type A (a serotonin deaminating enzyme). Radioenzyme assay of haemolymph from high-potassium injected flies shows that the amount of serotonin present could account for all of the retrievable bioactivity. Significant amounts of serotonin were found in the cerebral ganglion, the thoracic ganglion and nerves attached to the thoracic ganglion. Nerve sectioning experiments demonstrated that the abdominal nerves and the anterior nerves supplying the neck muscles are not involved in the normal salivatory response. However the cerebral-thoracic connective must be intact and it is suggested that release of serotonin is effected close to the main body of the thoracic ganglion. Some of the implications of the neurohormonal role of serotonin are discussed.

Brain and liver monoamine oxidase type A and type B activity in alcoholics and controls.

Monoamine oxidase activity in human postmortem brain and liver samples was measured in a group of patients with a prior history of alcoholism and compared to a control group with no prior history of alcoholism. Liver samples from patients with a prior history of alcoholism showed significantly lower monoamine oxidase activity with both phenylethylamine and serotonin as substrates. Postmortem brain samples, however, were not different in the two groups.

Saturated amines and diamines as substrates which inhibit beef liver mitochondrial monoamine oxidase.

Monoamines and diamines of 8-12 carbon atoms initially serve as substrates for purified beef liver monoamine oxidase but then lead to inhibition. The inhibition is not solely the result of aldehyde formation as addition of decylaldehyde does not inhibit benzylamine oxidation. Furthermore, neither the addition of alcohol dehydrogenase and NADH nor of semicarbazide prevent the inhibition of diaminodecane oxidation. The formation of a Schiff base on the enzyme surface resulting in aggregation or occlusion of the enzyme may be a cause of the inhibition. When concentrated enzyme solutions (greater than or equal to 1 mg/ml) are reduced by long-chain amines, 100% O2 causes only partial return of the flavin peak at 450 nm while enzyme activity continues to decrease. Substantial recovery of activity occurs (over a 3-4 week period) when inhibited enzyme is sedimented and resuspended in fresh buffer. These observations are discussed and compared with inhibition observed by other investigators with the substrate phenylethylamine.

Isolation of pure, catalytically active human liver monoamine oxidase B: Antibody complex

Monoamine oxidase B was purified from human liver mitochondria using a monoclonal antibody, MAO B-1C2, which recognizes monoamine oxidase B but not A. Triton X-100 extracts of mitochondria were incubated with purified MAO B-1C2 (IgG 1), and the catalytically active enzyme:antibody complex was isolated by affinity chromatography on Protein A-Sepharose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the complex revealed the presence of four polypeptide bands (monoamine oxidase B, 57,900 dalton; antibody heavy chain, 52,200 dalton; and two light chains, 29,400 and 27,700 dalton), and indicated a 1:1 stoichiometric ratio of enzyme to antibody. This method gave 154-fold purification of the enzyme from mitochondria.

Alterations in monoamine oxidase activity after single and repeated exposure of rats to chlorphentermine

Monoamine oxidase (MAO) activity was determined in rat tissues following in vivo treatments with chlorphentermine (CP). Oxidation of seven amine substrates by liver, lung or brain mitochondrial MAO was investigated at 2 h after a single i.p. injection (60 mg/kg) or after repeated injection (once daily for 3 days, 20 mg/kg, i.p.). Deamination of the type A substrates, norepinephrine, serotonin and octopamine, was decreased significantly in liver, lung and brain after both single and repeated injections. Oxidative deamination of tyramine and dopamine (type A + B substrates) was also lowered in all organs after single and repeated exposure to CP, but to a lesser degree than the type A substrates. However, oxidation of the type B substrates, benzylamine and tryptamine, was unaffected by CP administration in comparison to control. These data indicate that CP is a specific inhibitor of mitochondrial MAO, form A.

Cannabis extract, but not Δ1-tetrahydrocannabinol, inhibits human brain and liver monoamine oxidase

1. 1. Mitochondrial monoamine oxidase (MAO) of human brain and liver was inhibited by low concentrations of cannabis extract (CE) and a cannabinoid fraction isolated from it. 2. 2. Δ 1-Tetrahydrocannabinol (THC) did not elicit any inhibitory effect on the enzyme. 3. 3. The inhibition of MAO activity by CE and by its active fraction was more pronounced when the monoamine substrates 2-phenylethylamine (PEA) and benzylamine (BA) were used, as compared to the inhibition of the enzyme activity when 5-hydroxytryptamine was the substrate. 4. 4. The active cannabinoid fraction was found to be more potent than CE in inhibiting the activity of MAO with either substrate. 5. 5. The isolated fraction contains at least two cannabinoids with R f values of 0.67 and 0.71 on silica gel thin layer chromatography (TLC), as determined with toluene/chloroform/methanol (100:10:1, by volume) as the solvent system. 6. 6. The findings of this study emphasize the need for further exploration of the potential of cannabis as a source for therapeutic agents.

Enzymic and Molecular Characteristics of a New Form of Monoamine Oxidase, Distinct from Form-A and Form-B

The present study was undertaken to clarify the enzymic and molecular properties of monoamine oxidase (MAO) In carp brain. In particular, its sensitivities to selective MAO inhibitors, kinetic properties and molecular weight were compared with those of the enzyme in carp liver. The selective and potent MAO-A and MAO-B inhibitors FLA 788( + ), FLA 336( + ), MD 780236 and benzylcyanide caused dose-dependent inhibitions of MAO activity in both carp brain and liver; the inhibition curves were all single-sigmoidal, and the degrees of Inhibition of the activities towards 5-hydroxytryptamine (5-HT, selective MAO-A substrate), tyramine (substrate for both forms of MAO) and β-phenylethylamine (PEA, selective MAO-B substrate) were similar. This was also the case for inhibition of activity in carp brain by the irreversible and selective MAO-A and MAO-B inhibitors clorgyline and l-deprenyl, indicating the presence in both preparations of a single MAO which differs from either form of MAO. Studies on the substrate specificities and Km values for these three substrates and the inhibitory effects of some compounds suggested that the enzymic characters of MAO in carp preparations were similar and that these enzymes might be FAD-containing enzymes, like MAO in various mammals. By labelling the preparations with radioactive pargyline and then subjecting them to sodium dodecyl sulfate electrophoresis, the apparent molecular weights of carp brain and liver MAO were estimated as 60,000 daltons. The same value was also obtained for rat brain and liver mitochondrial MAO-B. These results indicate that by the present definitions of MAO-A and MAO-B, MAO in carp brain and liver is similar to, but distinct from, both these forms of MAO.

Enzymic and molecular characteristics of a new form of monoamine oxidase, distinct from form-A and form-B.

The present study was undertaken to clarify the enzymic and molecular properties of monoamine oxidase (MAO) in carp brain. In particular, its sensitivities to selective MAO inhibitors, kinetic properties and molecular weight were compared with those of the enzyme in carp liver. The selective and potent MAO-A and MAO-B inhibitors FLA 788(+), FLA 336(+), MD 780236 and benzylcyanide caused dose-dependent inhibitions of MAO activity in both carp brain and liver; the inhibition curves were all single-sigmoidal, and the degrees of inhibition of the activities towards 5-hydroxytryptamine (5-HT, selective MAO-A substrate), tyramine (substrate for both forms of MAO) and beta-phenylethylamine (PEA, selective MAO-B substrate) were similar. This was also the case for inhibition of activity in carp brain by the irreversible and selective MAO-A and MAO-B inhibitors clorgyline and I-deprenyl, indicating the presence in both preparations of a single MAO which differs from either form of MAO. Studies on the substrate specificities and Km values for these three substrates and the inhibitory effects of some compounds suggested that the enzymic characters of MAO in carp preparations were similar and that these enzymes might be FAD-containing enzymes, like MAO in various mammals. By labelling the preparations with radioactive pargyline and then subjecting them to sodium dodecyl sulfate electrophoresis, the apparent molecular weights of carp brain and liver MAO were estimated as 60,000 daltons. The same value was also obtained for rat brain and liver mitochondrial MAO-B. These results indicate that by the present definitions of MAO-A and MAO-B, MAO in carp brain and liver is similar to, but distinct from, both these forms of MAO.

ESR analysis of the FAD in bovine liver monoamine oxidase

Two hydrazine spin labels, 1-oxyl-2,2,5,5-tetramethylpyrroline-3-carbonyl ethyl hydrazine and 1-oxyl-2,2,6,6-tetramethylpiperidino-4-hydrazine, were synthesized as probes of the FAD binding site of monoamine oxidase. The reporter nitroxide moiety showed an ESR spectrum classified as partially immobilized which is indicative of FAD near the surface of the enzyme. Attempts to pick up flavin semiquinone or free radical intermediates during substrate oxidation with the spin traps 5,5-dimethyl-1-pyrroline-1-oxidase and phenyl-t-butylnitrone were not successful.

Comparative aspects of monoamine oxidase.

This short review attempts to summarize a small fraction of the comparative aspects of the flavoprotein enzyme, monoamine oxidase (MAO), which is found mainly in the outer mitochondrial membranes of a wide variety of animal species. It deaminates both aromatic and aliphatic monoamines in the form Open image in new window to their corresponding aldehydes and hydrogen peroxide. Some of the evidence is described which shows that MAO can be divided into two catalytically different forms, called MAO-A and MAO-B. This evidence derives from the fact that the substrate specificity of MAO depends not only on the species but also on the particular tissue studied. Some inhibitors of MAO including the irreversible inhibitors, clorgyline and deprenyl show considerable selectivity towards one or other form of the enzyme. Under appropriate conditions clorgyline is about 1000 times more active against MAO-A while deprenyl is more active against MAO-B. Use of these inhibitors will measure the proportions of the two forms of activity as they deaminate a fixed concentration of a chosen substrate. Such experiments may also reveal the presence of another deaminating enzyme that is not FAD-dependent and is not inhibited by clorgyline. With appropriate radioactive ligands it is possible to measure the number of active centres of each form and thus allow the relative turnover rate constants for MAO-A and MAO-B to be calculated for each substrate. As an example, the determination of MAO-A:B ratio is illustrated for rat and giraffe liver MAO. In addition a summary is given of the type of MAO activity responsible for the deamination of 5-hydroxytryptamine, tyramine, β-phenethylamine and benzylamine in heart, liver and brain of a few animal species.

Investigations of the mechanism of selective inhibition of type B mitochondrial monoamine oxidase by phosphatidylserine

Liposomes of phosphatidylserine (PS) were found to inhibit strongly the B-form of membrane bound monoamine oxidase (MAO) isolated from rat and bovine liver, while having no effect on the rat liver A-form. Use of 14C-liposomes demonstrated high levels of PS association with the membrane, which could not be removed by extensive washing with high ionic strength buffers. The inhibition of MAO-B was not reversed on further perturbation of the membrane by chaotropic agents, sonication, or treatment with additional liposome preparations of phosphatidylcholine or phosphatidylinositol. Partial reversal of the inhibition was found when the PS-treated bovine liver membrane was solubilizcd with the detergent octyl glucoside. PS, however, had no effect on a solubilized preparation of bovine liver MAO. These results suggest a specific interaction between MAO and PS rather than an indirect effect of bulk changes in membrane properties, but an intact membrane was. nevertheless, required to mediate the inhibition. Comparison of the decreases in apparent levels of MAO-B in rat liver mitochondrial membranes that were calculated from changes in relative catalytic activities with A and B specific substrates or changes in sensitivity to A-form specific reversible and irreversible inhibitors. all showed good quantitative correlation. Lineweaver-Burk plots of the effect of PS incorporation into bovine liver mitochondrial membranes on MAO oxidation of phenylethylamine exhibited the expected pattern for a noncompetitive inhibitor acting on a ping-pong mechanism bireactant enzyme. On the basis of these results, a possible in vivo role for the acidic phospholipids in regulating apparent levels of MAO from one tissue to another and/or in response to environmental effects is proposed.

Monoamine oxidase in pancreatic islets, exocrine pancreas, and liver from rats. Characterization with clorgyline, deprenyl, pargyline, tranylcypromine, and amezinium.

Monoamine oxidase (MAO) was characterized in tissue homogenates from pancreatic islets, exocrine pancreas, and liver from rats. Phenylethylamine was preferentially deaminated by pancreatic islet MAO while 5-hydroxytryptamine was preferentially deaminated by MAO from exocrine pancreas, and tyramine was a good substrate for both tissues. All three substrates were well deaminated by liver tissue. Clorgyline, a selective inhibitor of MAO-A, preferentially inhibited deamination of 5-hydroxytryptamine by all three tissue homogenates, while deprenyl, a selective inhibitor of MAO-B, preferentially inhibited deamination of phenylethylamine. In the case of pargyline, a less selective MAO-B inhibitor, the preference in favour of phenylethylamine was less pronounced. According to these results, MAO in pancreatic islets can be classified as predominantly type B enzyme species and MAO in exocrine pancreas as predominantly type A enzyme species while both types of the enzyme are present in the liver. Using the same three MAO substrates and compared with the effects of the selective enzyme inhibitors, clorgyline and deprenyl, tranylcypromine can be classified as a potent nonselective inhibitor of MAO in homogenates of all three tissues investigated with a slight preference in favour of the inhibition of the B-form of the enzyme, while in contrast amezinium can be classified as a weak nonselective inhibitor of MAO with a slight preference in favour of the inhibition of the A-form of the enzyme. All MAO inhibitors tested also inhibited insulin secretion by isolated incubated rat pancreatic islets, however only at IC50 which were two to three decimal powers higher than those necessary for the inhibition of the MAO activity, thus indicating that inhibition of MAO activity and inhibition of insulin secretion are apparently not closely related.

Effect of prenatal exposure to phenobarbital on the development of monoamine oxidase and glucocorticoids

1. 1. Fetal exposure to phenobarbital (PB, 30 mg/kg) 2–5 days prior to parturation was conducted in Sprague-Dawley rats. The fetal exposure to PB resulted in: (a) Significant increase ( P < 0.01) in body wt at 23 and 42 days of age; (b) Significant increase ( P < 0.01) in brain wt at all ages studied; (c) Significant decline ( P < 0.05) in liver monoamine oxidase at day 13 of age; (d) Significant ( P < 0.05) increase in plasma corticosterone levels at days 23 and 42.

A new type of mitochondrial monamine oxidase distinct from type-A and type-B

The characteristics of mitochondrial monoamine oxidase (MAO) in carp liver were studied with MAO inhibitors and substrates. This enzyme was thermolabile, but was stabilized in the presence of bovine serum albumin. With clorgyline and deprenyl, single-sigmoidal curves for inhibition of the activity towards tyramine or 5-hydroxytryptamine were obtained; the sensitivities to the two inhibitors were identical. The activity towards β-phenylethylamine was not completely inhibited by clorgyline or deprenyl, but the remaining activity was inhibited by semicarbazide and the inhibition curves by either clorgyline or deprenyl and semicarbazide were also identical to the curves with the other two substrates. These results suggest that carp liver mitochondria contain “classical” MAO and a clorgyline- and deprenyl-resistant amine oxidase and that the classical MAO does not seem to be MAO-A or MAO-B, which are present in mitochondria of most mammalian tissues.

Effects of aldehyde dehydrogenase inhibitors on enzymes involved in the metabolism of biogenic aldehydes in rat liver and brain

The effects of the aldehyde dehydrogenase inhibitors disulfiram, coprine and cyanamide on enzymes involved in the metabolism of biogenic aldehydes in rat liver and brain were studied. Both liver and brain aldehyde dehydrogenase activities were significantly decreased in rats pretreated with these drugs. In the liver, the low- K m aldehyde dehydrogenase activity was markedly decreased by all three drugs after 2 and 24 hr whereas only cyanamide inhibited the high- K m enzymes. The brain ALDH-activity with a low acetaldehyde concentration was significantly decreased by coprine and cyanamide at both times tested, whereas disulfiram caused no change after 2 hr but an inhibition of 38% after 24 hr. The brain ALDH-activity with a high acetaldehyde concentration was significantly decreased by coprine and cyanamide but not by disulfiram. The activity of the substrate specific enzyme succinate semialdehyde dehydrogenase in brain was slightly but significantly decreased in rats pretreated with cyanamide but not in rats pretreated with disulfiram or coprine. None of the drugs caused any changes in the activities of aldehyde reductase and monoamine oxidase in brains in vivo. The activity of monoamine oxidase in liver was significantly decreased by coprine after 24 hr. In contrast to the effects obtained in vivo, disulfiram was found to be an inhibitor in vitro of brain succinate semialdehyde dehydrogenase and liver monoamine oxidase. Aldehyde reductase was slightly inhibited by both disulfiram and 1-aminocyclopropanol in vitro.

The nature of the inhibition of rat liver monoamine oxidase types A and B by the acetylenic inhibitors clorgyline, l-deprenyl and pargyline

The kinetics of inhibition of rat liver mitochondrial monoamine oxidase by clorgyline, l-deprenyl and pargyline are consistent with a mechanism whereby a reversible interaction between the inhibitor and the enzyme active site under conditions of thermodynamic equilibrium is followed by a time-dependent formation of the covalently-bound enzyme-inhibitor adduct. The K i value for the reversible interaction between clorgyline and monoamine oxidase A is about 1000 times lower than that towards the B-form of the enzyme, and this difference is sufficient to account for most, but not all, of the selectivity of the inhibition caused by this compound. The K i value of the monoamine oxidase B selective inhibitor l-deprenyl towards that form of the enzyme is only about 40-fold lower than that towards the A-form. However, in this case, the rate of formation of the irreversible adduct is considerably faster for the B-form than for the A-form and this makes a major contribution to the selectivity of this compound. Pargyline shows a K i value towards monoamine oxidase B that is only 8 times lower than that towards the A-form and in this case the rates of formation of the enzyme-inhibitor adducts are similar. The significance of these results are discussed in terms of the selective inhibition of monoamine oxidase.

Glutamate — oxaloacetate transmainase, monoamine oxidase, diamine oxidase and mixed function oxidase activities in rat liver after portacaval shunt

1 month after portacaval shunt, some enzymatic activities of rat liver were tested and a significant increase of diamine oxidase was observed.

Inhibition of rabbit liver monoamine oxidase by nitro aromatic compounds

Nitrobenzenoid, nitroheterocyclic and cyanobenzoid compounds inhibit type B monoamine oxidase. A partially purified enzyme preparation from rabbit liver mitochondria, oxidizing p-dimethyl-aminobenzylamine as the substrate, was competitively inhibited by nitrobenzoid compounds with K i values in the range of 0.28 μM for p-dinitrobenzene to 0.56 mM for p-nitrobenzoic acid. The potencies of nitrobenzoid compounds were positively correlated with the Hammett sigma value for each substituent on nitrobenzene. Dinitro derivatives were slightly more potent than the corresponding mononitro compounds but not as potent as would be expected from their sigma values. For the nitroheterocyclic compounds, inhibition was also competitive; the lowest K i was 1.3 μM for 5-nitrofurfural semicarbazone (nitrofurazone). Compounds with cyano groups in place of nitro groups were also inhibitory; the most potent was p-acetobenzonitrile with a K i of 1.3 μM. The results of this study indicate that, in addition to nitrobenzoid compounds, other compounds with planar, electron-deficient nuclei are effective inhibitors of type B monoamine oxidase. Although hydrophobic and steric parameters may play some role in inhibition, the predominant factor is the electron-withdrawing power of the ring substituents.