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本研究利用结合到 BSA上的 deltorphin 肽 (DADT )免疫 Balb/c小鼠 ,成功地制备了特异性抗 DADT 单克隆抗体。交叉试验表明 ,所获抗体仅识别 DADT 和 DADT 而不识别其 L型同分异构体 L ADT ,也与其它 DADT 类似物无交叉反应。Epitope分析表明 ,该抗体识别 DADT 的 N末端特异氨基酸序列 ,其中包括对其活性有重要影响的第二位的 D-Ala-nine。利用这一抗体对成年大鼠脑组织和新生大鼠培养神经元进行了免疫组织化学研究。大鼠脑中有含量较低的 DADT 样免疫活性 ,主要分布在某些特定部位的神经纤维和神经元胞体 ,如终纹床核、杏仁核、海马、伏核、下丘脑外侧区、黑质、中央灰质等部位。然而在培养的大鼠脑神经元 ,DADT 样免疫活性的含量却很丰富 ,神经元的胞体和突起均呈较强的 DADT 样免疫活性。两项结果均提示哺乳动物脑神经元很可能产生一种与 DADT 在结构上极其类似的物质
In this study, Balb / c mice were immunized with deltorphin peptide (DADT) bound to BSA, and monoclonal antibodies specific for DADT were successfully prepared. Cross-tests showed that the antibodies obtained recognized only DADT and DADT without recognizing their L-isomer, LADT, and did not cross-react with other DADT analogues. Epitope analysis showed that the antibody recognizes the N-terminal specific amino acid sequence of DADT, including D-Ala-nine, which is second most important for its activity. The use of this antibody on adult rat brain and neonatal rat neurons were immunohistochemically studied. DADT-like immunocompetence was found in the rat brain, mainly distributed in nerve fibers and neuronal somatic cells in some specific sites, such as nucleus elaulentis, amygdala, hippocampus, nucleus accumbens, lateral hypothalamic area, substantia nigra , Central gray matter and other parts. However, the content of DADT-like immunocompetent neurons in cultured rat brain neurons is very rich. The neurons have strong DADT-like immunosuppressive activity in their somatic cells and neurites. Both results suggest that mammalian brain neurons are likely to produce a substance that is structurally very similar to DADT