Localization Of Monoamine Oxidase Research Articles

Localization of monoamine oxidase A and B and semicarbazide-sensitive amine oxidase in human peripheral tissues.

Monoamine oxidase (MAO) A and B and semicarbazide-sensitive amine oxidase (SSAO) localizations in peripheral human tissues were compared by immunohistochemistry. The primary antibodies used were mouse monoclonal anti-human MAO-A (6G11/E1) and anti-human MAO-B (3F12/G10/2E3) and a rabbit polyclonal anti-bovine SSAO antibody. Immunoreactivities of the samples, obtained from 6 routine autopsy cases, showed different distributions in the tissues studied (heart, lung, duodenum, liver, pancreas, spleen, thyroid gland, adrenal gland and kidney). The relative MAO-A, MAO-B and SSAO distributions indicated a widespread distribution of these enzymes in the human body that is characterized by a matching cellular pattern in only few tissues. These differences suggest that each amine oxidase may play a specific function in, at least some, peripheral tissues.

Localization of monoamine oxidase A and B in human pancreas, thyroid, and adrenal glands.

We studied monoamine oxidase (MAO) A and B localization in human pancreas, thyroid gland, and adrenal gland by immunohistochemistry. The primary antibodies used were mouse monoclonal anti-human MAO-A (6G11/E1) and anti-human MAO-B (3F12/G10/2E3). Samples were obtained from six routine autopsy cases and fixed in 2% paraformaldehyde. Exocrine pancreas showed a widespread distribution of MAO-A, whereas MAO-B was present only in centroacinar cells and epithelial cells of pancreatic ducts. In endocrine pancreas, MAO-A was observed in around 50% of islet cells, whereas MAO-B was less abundant and was restricted to the periphery of islets. Thyroid gland showed strong MAO-A immunoreactivity in all cell types and was MAO-B-negative. In adrenal gland, the capsule displayed MAO-A but not MAO-B immunoreactivity, whereas the cortex showed widespread MAO-A staining but was MAO-B-negative in interstitial cells. Finally, in the medulla only a few scattered cells showed either MAO-A or MAO-B immunoreactivity. To our knowledge, these data represent the first study of the cellular distribution of MAO-A and MAO-B in the three human tissues included.

Localization of monoamine oxidase A and B mRNA in the rat brain by in situ hybridization

Monoamine oxidases A and B (MAOA and MAOB) are the major catabolic isoenzymes of catecholamines and serotonin in the mammalian brain. Although the distribution of the monoamine oxidase protein has been mapped by ligand binding and immunohistochemistry, the sites of MAOA and MAOB synthesis have not been precisely determined. In this study, we used in situ hybridization to visualize MAOA and MAOB mRNAs in the rat brain by using specific cDNA and oligonucleotide probes. MAOA mRNA was localized in major monoaminergic cell groups, such as the dorsal vagal complex, the C1/A1 groups, the locus ceruleus, the raphe nuclei, the substantia nigra, and the ventral tegmental area. MAOA mRNA was also found in forebrain structures, such as the cortex, the hippocampus, the thalamus, and the hypothalamus. In contrast to the distribution of MAOA mRNA, high levels of MAOB mRNA were present in only three brain regions: the area postrema, the subfornical organ, and the dorsal raphe. The in situ visualization of MAO mRNA demonstrates that MAOA mRNA synthesis is wide spread in many catecholaminergic and serotonergic cell groups, whereas MAOB mRNA synthesis is far more discrete and limited. The different expression patterns of MAOA and MAOB suggests that may also have different physiological functions. Synapse 25:30–36, 1997. © 1997 Wiley-Liss, Inc.

Localization of monoamine oxidase A and B mRNA in the rat brain by in situ hybridization.

Monoamine oxidases A and B (MAOA and MAOB) are the major catabolic isoenzymes of catecholamines and serotonin in the mammalian brain. Although the distribution of the monoamine oxidase protein has been mapped by ligand binding and immunohistochemistry, the sites of MAOA and MAOB synthesis have not been precisely determined. In this study, we used in situ hybridization to visualize MAOA and MAOB mRNA in the rat brain by using specific cDNA and oligonucleotide probes. MAOA mRNA was localized in major monoaminergic cell groups, such as the dorsal vagal complex, the C1/A1 groups, the locus ceruleus, the raphe nuclei, the substantia nigra, and the ventral tegmental area. MAOA mRNA was also found in forebrain structures, such as the cortex, the hippocampus, the thalamus, and the hypothalamus. In contrast to the distribution of MAOA mRNA, high levels of MAOB mRNA were present in only three brain regions: the area postrema, the subfornical organ, and the dorsal raphe. The in situ visualization of MAO mRNA demonstrates that MAOA mRNA synthesis is wide spread in many catecholaminergic and serotonergic cell groups, whereas MAOB mRNA synthesis is far more discrete and limited. The different expression patterns of MAOA and MAOB suggests that may also have different physiological functions.

Type‐specific localization of monoamine oxidase in the enteric nervous system: Relationship to 5‐hydroxytryptamine, neuropeptides, and sympathetic nerves

The localization in the guinea pig enteric nervous system (ENS) of monoamine oxidase (MAO) types A and B was investigated at the light and electron microscopic levels. Immunocytochemistry was used to visualize the enzyme protein and histochemistry was employed to study catalytic activity. Type specificity was achieved in histochemical studies by using deprenyl (0.5 microM) to inhibit MAO-B or clorgyline (0.1 microM) to inhibit MAO-A. The distribution of MAO-B immunoreactivity in the ENS corresponded to that of the sites of MAO activity found histochemically to be inhibited by deprenyl, but not clorgyline. MAO-B was observed to be the primary type of MAO found in the intrinsic elements of the ENS and was located in subsets of neurons in both submucosal and myenteric plexuses. MAO-B was not demonstrated immunocytochemically or histochemically in enteric glia, nor, at the light microscopic level, was there significant MAO-B activity or immunoreactivity in serotonin (5-HT)-immunoreactive neuronal cell bodies. In the submucosal plexus about 50% of the neurons expressed MAO-B; these neurons also contained neuropeptide y (NPY) and/or calcitonin gene related peptide (CGRP), but not substance P or vasoactive intestinal polypeptide (VIP). About 10% of myenteric neurons were intensely reactive for MAO-B; again MAO-B was co-localized with NPY and/or CGRP. In contrast to intrinsic neurons, extrinsic CGRP-immunoreactive nerve fibers contained no demonstrable MAO activity or immunoreactivity. Moreover, the sympathetic innervation, identified as varicose axons that degenerated after administration of 6-hydroxydopamine, contained abundant MAO-A, but no MAO-B activity or immunoreactivity. It is concluded that MAO-B is characteristic of a subset of intrinsic enteric neurons, while MAO-A is confined to the sympathetic innervation, which is extrinsic. At the electron microscopic level individual cells varied greatly in their degree of immuno- or cytochemically demonstrable MAO-B, which was most concentrated on the outer membranes of mitochondria. MAO-B immunoreactivity (but not cytochemical activity) was found on mitochondria in some serotoninergic perikarya identified by the simultaneous radioautographic detection of the uptake of 3H-5-HT. Mitochondria in most serotoninergic axon terminals displayed both MAO-B activity and immunoreactivity. Neurons receiving serotoninergic synapses often, but not invariably, contained MAO-B. Inhibition of neither MAO-B nor MAO-A appeared to slow the disappearance of 3H-5-HT loaded into enteric neurons significantly, even when intraneuronal storage of 5-HT was inhibited with tetrabenazine.(ABSTRACT TRUNCATED AT 400 WORDS)

Monoamine oxidase-containing nerve fibers in the major cerebral arteries of rats

The localization of monoamine oxidase (MAO) in nerve fibers associated with the major cerebral arteries in rats was studied using a new coupled peroxidation method modified by adding nickel ammonium sulfate at the electron microscopic level. MAO was localized in some unmyelinated axons, both in the adventitia and in the periadventitial nerve bundles. Schwann cell cytoplasm encircling myelinated axons in the periadventitial nerve bundles also contained a MAO-reactive substance. The incidences of MAO-containing axons in the adventitial layer of the anterior cerebral, middle cerebral, internal carotid and basilar arteries were 32.3%, 29.5%, 29.6% and 21.1%, respectively. In the periadventitial nerve bundles, MAO activity was also demonstrated in 10.8% among unmyelinated axons. Preincubation with clorgyline, a specific inhibitor of MAO A, suppressed staining in the axons, both in adventitia and in periadventitial nerve bundles, but not in the Schwann cell cytoplasm. Conversely, preincubation with deprenyl, is regarded as MAO A and MAO in the Schwann cell cytoplasm as MAO B. In immunosympathectomized rats (anti-NGF-treated rats), MAO reactivity was suppressed in axons associated with cerebral arteries, but was retained in some Schwann cell cytoplasm. The results indicate that the MAO-containing unmyelinated axons coincide with the postganglionic noradrenergic ones. Thus, histochemical MAO staining may be utilized to study postganglionic sympathetic nerve fibers presumably innervating the major cerebral arteries at the electron microscopic level.

Localization of monoamine oxidase B in human brain by autoradiographical use of11C-labelled L-deprenyl

11C-labelled L-deprenyl in vitro autoradiography was used to study the regional distribution of MAO-B in human brain. 80 microns thick cryosections from two human brains, a 67 years old female and a 58 years old male, were taken on tape/paper and transferred on to a gelatinized glass plate. The sections were then incubated with 34 and 54 nM 11C-L-deprenyl for 15 min and exposed to a film sensitive to high energy radiation for 2 hours. The autoradiograms obtained were analyzed by computerized densiotometry. High 11C-deprenyl binding was found in the caudate nucleus, putamen, thalamus, substantia nigra, medial and lateral geniculate bodies, hippocampus and periaqueductal gray. Moderate to low binding was observed in cerebral cortex. Cerebral cortex and white matter showed the lowest binding. The autoradiographic technique described proved to be a fast and reliable method to investigate the topographic localization of MAO-B in large cryosections of human brain.

Localization of monoamine oxidase A and B genes on the mouse X chromosome

Localization of monoamine oxidase A and B genes on the mouse X chromosome

Intra- and extra-dopamine-synaptosomal localization of monoamine oxidase in striatal homogenates from four species

MAO-A and MAO-B activities within and outside dopaminergic synaptosomes in homogenates of striatal tissue from pig, cat, rat and human brains have been studied by using a specific “low substrate concentration technique” with dopamine. It was found that within the synaptosomes, MAO-A activity predominated in all species. Outside the synaptosomes there were more pronounced differences and only in the rat did MAO-A predominate, while in the other species MAO-B predominated. When estimating MAO-A and -B activities with a conventional method the activity of MAO-B predominated in man, cat and pig. Thus, also in species where the MAO-B activity (as estimated in a conventional way) was dominating, the intrasynaptosomal deamination of dopamine was brought about mainly by MAO-A. The “low substrate concentration technique”, more adequately reflects physiological conditions by taking into account the higher concentration of monoamine transmitter substrates within the monoamine neurons. With this technique it was found that in all species (with the possible exception of man) the oxidation rate was higher within than that oustide the DA-synaptosomes. In man the unavoidable longer time between death and estimation of the enzyme activity may be the cause of the deviating result.

The vectorial orientation of human monoamine oxidase in the mitochondrial outer membrane.

1. The localization of monoamine oxidase in the mitochondrial outer membrane was studied in preparations of human liver mitochondrial and brain-cortex non-synaptosomal and synaptosomal mitochondria. 2. Immunochemical accessibility in iso-osmotic and hypo-osmotic mitochondrial preparations was used to localize the enzyme. 3. It was shown that the immunochemically accessible tyramine-oxidizing activity was distributed approximately equally on both surfaces of the membrane in human liver and brain-cortex non-synaptosomal mitochondria. However, the immunochemically accessible beta-phenethylamine-oxidizing activity was situated predominantly on the outer surface, and the immunochemically accessible 5-hydroxytryptamine-oxidizing activity was situated predominantly on the inner surface of the mitochondrial outer membrane in liver and brain-cortex non-synaptosomal mitochondrial preparations. 4. Considerable variation in the distribution of the enzyme in preparations of synaptosomal mitochondria was seen. 5. The simplest model consistent with our observations is that, in liver and brain-cortex non-synaptosomal mitochondria, the tyramine-oxidizing activity is distributed on both sides of the mitochondrial outer membrane, the beta-phenethylamine-oxidizing activity is located on the outer surface of the outer membrane and the 5-hydroxytryptamine-oxidizing activity is located on the inner surface of the mitochondria outer membrane.

A coupled peroxidatic oxidation technique for the histochemical localization of monoamine oxidase A and B and benzylamine oxidase.

A coupled peroxidatic oxidation technique is presented which employs benzylamine and tyramine as substrates and clorgyline, deprenyl, phenelzine and pargyline as specific inhibitors. Using this technique with frozen sections of human term placenta and rat liver, the histochemical localization of monocamine oxidase A and B and bnezylamine oxidase has been demonstrated.

Subcellular localization of monoamine oxidase in bovine thalamus tissue using immunoferritin conjugates.

A combination of discontinuous sucrose gradient analysis and polyacrylamide electrophoresis was used to isolate one of the multiple forms of monoamine oxidase (MAO) from bovine thalamus. This substance (the principle form and the most anodic of the five MAO forms observed) was used as a basis for an immunoferritin-electron microscope approach to determine the subcellular localization of MAO in thalamus. This form of MAO, as well as antigenically related forms, was found to reside mainly on the outer mitochondrial membrane. In addition, the action of SDS on solubilization and interconversion of MAO forms was studied and found to be dependent on the concentration and time of reaction of SDS with thalamus tissue.

Electron microscopic localization of monoamine oxidase in the rat liver.

Two methods have been employed to localize monoamine oxidase activity in the cells of rat liver, using either 2-(2'-benzothiazolyl)-5-stryl-3-(4'-phtalhydrazidyl) tetrazolium chloride (BSPT) or ferricyanide as electron acceptor. With both methods monoamine oxidase activity was found both in the inner and the outer mitochondral membrane, although the outer membrane appeared the most probable location. In addition the BSPT method but not the ferricyanide method, revealed monoamine oxidase activity in the endoplasmatic reticulum. The results obtained by the two methods have been compared and are discussed in view of available biochemical data on monoamine oxidase.

Investigation of localization of monoamine oxidase (EC 1.4.3.4) in subfraction of rat liver mitochondria.

Subfractionation of the purified preparations of mitochondria was performed according to Schnaitman's digitonin method. In some experiments, polyvinyl sulphate (PVS) was added to the medium during the preparation and subfractionation of mitochondria. The formation of the "fluffy layer" was not observed in the presence of PVS. The "fluffy layer" was either removed or left within the pellet during the separation of the subfraction of mitoplasts from the supernate containing the outer membrane as well as the inter-membrane space. The monoamine oxidase (MAO) activity was determined by means of our own modification of Tabor's method. In this way the influence of aldehyde oxidase upon the obtained results could be eliminated. A part of MAO activity was found in the subfraction of mitoplasts both in the presence and absence of PVS in the medium. The obtained results suggest double localization of MAO both in the outer and inner membrane. The influence of the method of determination of MAO activity on the evaluation of its intra-mitochondrial activity has been discussed.

Histochemical demonstration of monoamine oxidase in the hypothalamo-hypophysial system of the tree sparrow and the rat.

Localization of monoamine oxidase (MAO) was investigated essentially according to the method of Glenneret al. (1957) in the hypothalamo-hypophysial system of the tree sparrow and the rat. The hypothalamic neurosecretory cells of both species showed relatively weak MAO activity. A similar localization of MAO activity was observed in the median eminence of both species: (1) slight or no MAO activity was observed in the ependymal layer, (2) relatively strong activity was revealed in the tissue just beneath the ependymal layer, (3) strong activity was revealed in the outer layer, particularly in the tissues surrounding capillary loops of the primary plexus. It is suggested that an adrenergic mechanism functions in the median eminence. In the pars nervosa, strong reaction was observed in the rat, while a weak reaction occurred in the tree sparrow. However, the color and the size of formazan crystals deposited in the rat pars nervosa differed from those in the hypothalamus. As a whole, the distribution of the neurosecretory material differed from the localization of MAO activity in the hypothalamo-hypophysial system. It is discussed that the neurosecretory neuron is not adrenergic but cholinergic.

COMPARATIVE STUDIES IN THE CHARACTERIZATION OF MONOAMINE OXIDASES.

SummaryFractionation of rat liver homogenates suggests a dual intracellular localization of monoamine oxidase, with the major portion in the mitochondria and a smaller but significant portion sedimenting with the microsomal fraction. Comparative studies on substrate specificities and Michaelis constants of monoamine oxidase preparations from liver, kidney and heart of rat and guinea pig indicate marked tissue differences in monoamine oxidases. Multiple substrate studies and other data support the view that each tissue contains a single, distinctive major monoamine oxidase.

Histology and cytochemistry of human skin. 20. The distribution of monoamine oxidase.

1. Using the method of Glenner et al. (8), we have demonstrate abundant monoamine oxidase in human skin. 2. In the epidermis, the reaction occurs in the cells of the malpighian layer. In all hair follicles, regardless of their states of activity, the wall of the pilosebaceous canal is intensely reactive. In growing follicles, the cells of the outer root sheath contain moderate enzyme activity particularly concentrated in the cells at the periphery, in the region above the bulb; small reactive granules are sparsely distributed in the cells of the dermal papilla. In quiescent follicles, there is moderate activity in the cells of the epidermal sac; the hair germ and the dermal papilla have a scant reaction. In the sebaceous glands, the duct and the undifferentiated cells at the periphery of the acini have a very strong reaction. Both apocrine and eccrine glands show strong enzyme activity. 3. Many cutaneous nerves contain monoamine oxidase. Reactive nerves form plexuses just beneath the epidermis, at the level of the sebaceous gland, and midway along the depth of the hair follicle; reactive nerves are also found around both eccrine and apocrine glands. Eccrine glands are surrounded by a richer bed of nerves than the apocrine glands are. The nerves to the arteries also contain the enzyme. 4. The localization of monoamine oxidase in human skin resembles that of succinic dehydrogenase; the cutaneous nerve fibers and the myoepithelial cells, which contain no succinic dehydrogenase, do, however, contain monoamine oxidase. 5. Many cutaneous nerve fibers, sensory and motor, seem to be reactive for both monoamine oxidase and cholinesterase, We do not know what this means in terms of physiology.

Physiological implications of the histochemical localization of monoamine oxidase.

Physiological implications of the histochemical localization of monoamine oxidase.