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1.
J Histochem Cytochem ; 48(1): 147-51, 2000 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10653595

RESUMO

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.


Assuntos
Glândulas Suprarrenais/enzimologia , Monoaminoxidase/isolamento & purificação , Pâncreas/enzimologia , Glândula Tireoide/enzimologia , Idoso , Idoso de 80 Anos ou mais , Especificidade de Anticorpos , Feminino , Humanos , Imuno-Histoquímica , Ilhotas Pancreáticas/enzimologia , Isoenzimas/imunologia , Isoenzimas/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Monoaminoxidase/imunologia
2.
Placenta ; 23(2-3): 163-71, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-11945082

RESUMO

A feature of pre-eclampsia is that circulating levels of maternal serotonin (5-hydroxytryptamine) are elevated and placental monoamine oxidase-A (MAO-A) activity, the major factor in the regulation of serotonin levels in pregnancy, is reduced. It is not known whether this is due to a reduced MAO-A protein content or a reduced catalytic turnover of the serotonin by MAO-A; this question has been addressed in the present work. Term placentae from normotensive and pre-eclamptic women were analysed for MAO-A specific mRNA expression (by semi-quantitative RT-PCR), MAO-A protein (by immunohistochemistry and quantitative ELISA, using a MAO-A specific monoclonal antibody), together with MAO activity (using [(3)H] labelled 5-hydroxytryptamine as substrate). Immunohistochemical analysis of placentae from both normotensive and pre-eclamptic women demonstrated that MAO-A protein is located in the cytoplasm of the placental syncytiotrophoblast layer, consistent with a mitochondrial location; no MAO-A protein was found in the nucleus. No MAO-B protein was detected in this placental layer, despite the presence of MAO-B mRNA. The results indicate that both total protein/g fresh weight and MAO-A protein/g fresh weight were approximately 40 per cent lower in pre-eclamptic than in normotensive placentae, but that there was no statistical difference in the expression of MAO-A mRNA in relation to GAPDH or actin mRNA or in MAO-A protein/mg total protein. However, MAO-A activity/g fresh weight was significantly reduced in pre-eclamptic placentae, in agreement with previous findings. This was found to be due to a 60 per cent reduction (P< 0.05) in the catalytic turnover (activity/molecule) of the enzyme. This study has therefore clearly shown that the expression of placental MAO-A specific mRNA and MAO-A protein are not specifically affected in pre-eclampsia, but that the catalytic efficiency of the expressed MAO-A enzyme in pre-eclamptic placentae is greatly reduced.


Assuntos
Monoaminoxidase/metabolismo , Pré-Eclâmpsia/enzimologia , Gravidez/metabolismo , Trofoblastos/enzimologia , Actinas/genética , Actinas/metabolismo , Adulto , Ensaio de Imunoadsorção Enzimática , Feminino , Gliceraldeído-3-Fosfato Desidrogenases/genética , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Humanos , Técnicas Imunoenzimáticas , Monoaminoxidase/genética , Oligonucleotídeos Antissenso/química , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
4.
Cell Tissue Res ; 313(3): 291-300, 2003 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-12898212

RESUMO

The distributions of monoamine oxidase (MAO)-A and -B proteins and mRNAs in human heart, lung, liver, duodenum, kidney and vasculature were compared using immunohistochemistry and cRNA in situ hybridisation. MAO-A and -B mRNA were detected in all tissues, to differing extents, but particularly in glomeruli, hepatocytes, and the crypts, muscularis mucosa and muscularis externa of duodenum. Renal proximal and distal tubules and loops of Henle had more intense labelling for mRNA of MAO-B than MAO-A; this was reflected in MAO protein expression. Little immunoreactivity was detected in glomeruli. Hepatocytes expressed MAO-A moderately, but MAO-B strongly. In lungs, similar moderately intense labelling for both MAO mRNAs and immunoreactivities was evident in pneumocytes, and epithelial and smooth muscle cells. Cardiomyocytes contained both MAO isoforms, but with more, albeit moderate, labelling for MAO-A. Both isoforms were expressed equally in duodenal villi, crypts, muscularis externa and mucosa; lower level expression occurred in mucosal and submucosal cells. MAO-A and -B mRNA were detected in endothelia, adventitia and media of a renal interlobular artery, but protein immunoreactivities were chiefly in the adventitia. The data reveal widespread tissue distribution of MAO mRNAs and proteins, but indicate that presence of MAO mRNAs does not invariably reflect quantitatively its protein expression.


Assuntos
Monoaminoxidase/metabolismo , Proteínas/metabolismo , RNA Mensageiro/metabolismo , Idoso , Idoso de 80 Anos ou mais , Vasos Sanguíneos/citologia , Vasos Sanguíneos/enzimologia , Vasos Sanguíneos/metabolismo , Duodeno/citologia , Duodeno/enzimologia , Duodeno/metabolismo , Feminino , Hepatócitos/enzimologia , Hepatócitos/metabolismo , Humanos , Imuno-Histoquímica , Hibridização In Situ , Isoenzimas/genética , Isoenzimas/metabolismo , Rim/irrigação sanguínea , Rim/enzimologia , Rim/metabolismo , Fígado/citologia , Fígado/enzimologia , Fígado/metabolismo , Pulmão/citologia , Pulmão/enzimologia , Pulmão/metabolismo , Masculino , Monoaminoxidase/genética , Miocárdio/citologia , Miocárdio/enzimologia , Miocárdio/metabolismo , Artéria Renal/enzimologia , Artéria Renal/metabolismo , Distribuição Tecidual
5.
Cell Tissue Res ; 304(2): 215-20, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11396715

RESUMO

We studied the localization of monoamine oxidase (MAO) A and B in human heart, liver, duodenum, blood vessels and kidney 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. All cardiomyocytes and hepatocytes showed MAO-A and MAO-B immunoreactivity. In the duodenum, both immunoreactivities were present in all cells of the villi, Lieberkühn crypts, muscularis mucosae and muscular layers, whereas Brunner glands were devoid of MAO-A and MAO-B staining. Endothelial cells of lymphatic vessels showed MAO-A but no MAO-B immunoreactivity, whereas arteries and veins presented MAO-A and MAO-B staining in muscular layers and fibroblasts but not in endothelial cells. In the kidney, renal tubuli showed MAO-A and MAO-B immunoreactivities, whereas collecting ducts and the Bowman's capsule showed only MAO-A staining. These data represent the first study of the cellular distribution of MAO-A and MAO-B in these human tissues. They show that both enzymes have a widespread distribution in the human body with a matching pattern in many, but not all tissues, and with strong differences from the pattern of distribution in rodents.


Assuntos
Monoaminoxidase/análise , Idoso , Idoso de 80 Anos ou mais , Especificidade de Anticorpos , Vasos Sanguíneos/citologia , Vasos Sanguíneos/enzimologia , Duodeno/citologia , Duodeno/enzimologia , Feminino , Humanos , Imuno-Histoquímica , Rim/citologia , Rim/enzimologia , Fígado/citologia , Fígado/enzimologia , Masculino , Pessoa de Meia-Idade , Monoaminoxidase/imunologia , Miocárdio/citologia , Miocárdio/enzimologia , Distribuição Tecidual
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