Auda G R, Kirk S H, Billett M A, Billett E E
Department of Life Sciences, Faculty of Science and Mathematics, Nottingham Trent University, Nottingham, United Kingdom.
J Histochem Cytochem. 1998 Dec;46(12):1393-400. doi: 10.1177/002215549804601208.
Monoamine oxidase (MAO) oxidatively deaminates vasoactive and biogenic amines and exists in two distinct forms (A and B), coded for by separate genes, which exhibit distinct substrate specificities and inhibitor sensitivities. Using specific primers for MAO-A and MAO-B mRNA in a reverse transcription-polymerase chain reaction (RT-PCR) on RNA from human liver, the predicted products for both enzymes were detected. Furthermore, RT-PCR on RNA from human placenta, believed to contain predominantly (or only) MAO-A protein, also indicated the presence of both A and B gene transcripts. The cellular distribution of MAO mRNA in placental tissue was analyzed by in situ hybridization of MAO-A and MAO-B mRNA-specific cRNA probes on paraffin sections. MAO-A mRNA was mainly evident in the syncytiotrophoblastic layer. None was detected in the vascular endothelium/smooth muscles. Significantly, MAO-B mRNA signal was also evident in the placental villi, notably in the syncytiotrophoblasts, intermediate trophoblasts, cytotrophoblasts, and the vascular endothelium. To our knowledge, this is the first demonstration of the cellular distribution of MAO mRNA in human placenta via in situ hybridization. The expression of MAO-B in placental tissue rather than in blood elements within placenta is also unequivocally demonstrated. These highly specific cRNA probes can now be used to study the distribution of MAO-A and MAO-B expression in other tissues.
单胺氧化酶(MAO)可氧化脱氨血管活性胺和生物胺,它以两种不同形式(A和B)存在,由不同基因编码,具有不同的底物特异性和抑制剂敏感性。使用针对MAO - A和MAO - B mRNA的特异性引物,对来自人肝脏的RNA进行逆转录 - 聚合酶链反应(RT - PCR),检测到了两种酶的预期产物。此外,对来自人胎盘的RNA进行RT - PCR,胎盘被认为主要(或仅)含有MAO - A蛋白,结果也表明存在A和B两种基因转录本。通过在石蜡切片上对MAO - A和MAO - B mRNA特异性cRNA探针进行原位杂交,分析了胎盘组织中MAO mRNA的细胞分布。MAO - A mRNA主要在合体滋养层中明显可见,在血管内皮/平滑肌中未检测到。值得注意的是,MAO - B mRNA信号在胎盘绒毛中也很明显,尤其是在合体滋养层细胞、中间滋养层细胞、细胞滋养层细胞和血管内皮中。据我们所知,这是首次通过原位杂交证明人胎盘中MAO mRNA的细胞分布。同时也明确证明了MAO - B在胎盘组织中的表达,而非在胎盘内的血液成分中。这些高度特异性的cRNA探针现在可用于研究MAO - A和MAO - B在其他组织中的表达分布。