Identification of an opioid peptide secreted by rat embryonic mixed brain cells as a promoter of macrophage migration
Identification of an opioid peptide secreted by rat embryonic mixed brain cells as a promoter of macrophage migration
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DOI：
10.1046/j.1460-9568.2000.00145.x
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发表时间：
2000-08-01
影响因子：
3.4
通讯作者：
Glowinski, J
中科院分区：
文献类型：
--
作者：
Calvo, CF;Cesselin, F;Glowinski, J
来自大鼠大脑的胚胎混合细胞的条件培养基被用于趋化性测定，以寻找潜在的趋化活性，这可以解释巨噬细胞前体对发育中的中枢神经系统（CNS）的浸润。在 E17 混合脑细胞 (E17-CM) 的条件培养基中发现了最有效的趋化活性。根据棋盘分析，该活性被证明是趋化性的而不是趋化性的。这种化学吸引力并不局限于脑巨噬细胞（BM），因为它在骨髓来源的巨噬细胞上也很明显。肽化合物对热和酸处理的耐受性以及对氨肽酶 M 消化的敏感性表明了肽化合物在此活性中的作用。与肽的阿片性质一致，不仅纳洛酮，而且 δ 阿片受体拮抗剂 ICI-174 也将响应 E17-CM 的 BM 迁移减少了 60%。当使用百日咳毒素处理的BM时，这种迁移活动不再有效。当将选择性阿片类激动剂的趋化作用与 E17-CM 的趋化作用进行比较时，δ 激动剂 DPDPE 吸引 BM 的效率最高。逆转录聚合酶链反应 (RT-PCR) 分析表明，δ 以及其他已知的阿片受体在 BM 和 E17 混合脑细胞中都有表达。最后，通过 RIA 在 E17-CM 中发现了甲硫氨酸脑啡肽样反应性。总而言之，这些观察结果表明，从胚胎混合脑细胞中释放的 δ 样阿片肽可能是巨噬细胞前体对发育中中枢神经系统的浸润的原因。
Conditioned media from embryonic mixed cells from the rat brain were used in a chemotaxis assay to look for potential chemotactic activity which could account for the infiltration of the developing central nervous system (CNS) by macrophage precursors. The most potent chemotactic activity was found in the conditioned medium from E17 mixed brain cells (E17-CM). Based upon checkerboard analysis, this activity was shown to be chemotactic rather than chemokinetic. This chemoattraction was not restricted to brain macrophages (BM) because it was as pronounced on bone marrow-derived macrophages. The implication of a peptide compound in this activity was suggested by its resistance to heat as well as acid treatments, and by its sensitivity to aminopeptidase M digestion. In agreement with the opioid nature of the peptide, not only naloxone, but also the delta opioid receptor antagonist ICI-174 reduced the migration of BM in response to E17-CM by 60%. This migratory activity was no longer effective when pertussis toxin-treated BM were used. When the chemotactic effects of selective opioid agonists were compared to that of E17-CM, DPDPE, the delta agonist, was the most efficient in attracting BM. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis indicated that delta as well as other known opioid receptors were expressed in both BM and E17 mixed brain cells. Finally, a Met-enkephalin-like reactivity was found by RIA in the E17-CM. Altogether, these observations suggest that a delta-like opioid peptide released from embryonic mixed brain cells could be responsible for the infiltration of the developing CNS by macrophages precursors.