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Identification of endogenous morphine and a mu3-like opiate alkaloid receptor in human brain tissue taken from a patient with intractable complex partial epilepsy

机译:难治性复杂性部分癫痫患者的人脑组织中内源性吗啡和mu3样鸦片生物碱受体的鉴定

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Background We set out to detect whether morphine is present in tissue taken from a patient with intractable temporal lobe epilepsy and to characterize the presence and nature of mu opiate receptor subtypes in this tissue. Material and Method In temporal lobe tissue, resected during anteromedial temporal lobectomy for intractable focal epilepsy, morphine was identified by quantitative radioimmunoassay (RIA) coupled to electrochemical detection via high-pressure liquid chromatography (HPLC). In addition, RNA isolated from the medial and lateral temporal lobe specimens was analyzed by conventional and real time reverse transcriptase-polymerase chain reaction (RT-PCR) for the expression of different human receptor gene transcripts. RIA revealed the presence of morphine at 3.4 nanograms per gram of tissue wet weight. Using RT-PCR and a primer specifically set for the mu3 (550 base pair fragment) and mu4 (880 base pair fragment) MOR splice variants, a mu4 splice variant was identified in both brain sections. Results Conclusions This human brain tissue study of a subject with temporal lobe epilepsy documents the presence of endogenous morphine and of a mu4 splice variant. These findings may have implications for our understanding of the mechanism of temporal lobe epilepsy.
机译:背景技术我们着手检测在患有顽固性颞叶癫痫患者的组织中是否存在吗啡,并表征该组织中鸦片受体亚型的存在和性质。材料和方法在颞叶前叶切除术中切除顽固性局灶性癫痫的颞叶组织中,通过定量放射免疫测定(RIA)与高压液相色谱(HPLC)电化学检测相结合,鉴定出吗啡。此外,通过常规和实时逆转录酶-聚合酶链反应(RT-PCR)分析了从颞叶内侧和外侧颞叶标本中分离出的RNA,以表达不同的人类受体基因转录本。 RIA显示吗啡的存在量为每克组织湿重3.4纳克。使用RT-PCR和专门为mu3(550个碱基对的片段)和mu4(880个碱基对的片段)MOR剪接变体设置的引物,在两个脑部均鉴定出一个mu4剪接变体。结果结论这项对颞叶癫痫患者的人脑组织研究表明存在内源性吗啡和mu4剪接变体。这些发现可能对我们对颞叶癫痫机制的理解有影响。

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