EXPRESSION OF TYPE-I AND TYPE-II INTERLEUKIN-1 RECEPTORS IN MOUSE-BRAIN
EXPRESSION OF TYPE-I AND TYPE-II INTERLEUKIN-1 RECEPTORS IN MOUSE-BRAIN
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DOI：
10.1016/0169-328x(94)90185-6
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发表时间：
1994-11-01
期刊：
影响因子：
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通讯作者：
KELLEY, KW
中科院分区：
文献类型：
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作者：
PARNET, P;AMINDARI, S;KELLEY, KW
尽管已在小鼠大脑中鉴定出 IL-1 的结合位点，但仍不清楚这些结合位点是否对应于 I 型或 II 型 IL-1 受体。使用定量放射自显影术确认 DBA/2 小鼠大脑中放射性标记的重组人 IL-1 α (I-125-HuIL-1 α) 特异性结合位点的存在。 IL-1 结合在齿状回中最高，由一类高亲和力结合位点组成，K-d 为 0.1 nM，B-max 为 57 fmol/mg 蛋白质。使用来自整个海马体的分离膜获得了类似的 0.2 nM K-d，尽管结合位点的数量较低（2 fmol/mg 蛋白质）。 I-125-HU-IL-1 α 与海马膜的亲和交联揭示了两种类型的 IL-1 受体蛋白的存在，与 I 型 (85 kD) 和 II 型 (60 kD) IL-1 受体的大小一致。然后合成寡核苷酸探针并用于 RT-PCR，随后进行 Southern blotting，以显示全脑表达 I 型和 II 型 IL-1 受体的转录本。小鼠神经母细胞瘤细胞系 C1300 表达 I 型而非 II 型 IL-1 受体 mRNA。 I 型受体蛋白可以通过流式细胞术在 C1300 神经元细胞系的膜上使用大鼠抗小鼠 I 型 IL-1 受体 MoAb 的间接免疫荧光进行鉴定。这些数据表明小鼠大脑表达 I 型和 II 型 IL-1 受体 mRNA 和蛋白质，并进一步支持 I 型 IL-1 受体由神经元合成和表达的观点。
Although binding sites for IL-1 have been identified in the mouse brain, it is still unknown whether these binding sites correspond to the type I or type II IL-1 receptor. Quantitative autoradiography was used to confirm the presence of specific binding sites for radiolabelled recombinant human IL-1 alpha (I-125-HuIL-1 alpha) in the brain of DBA/2 mice. IL-1 binding was highest in the dentate gyrus, consisting of a single class of high affinity binding sites with a K-d of 0.1 nM and a B-max of 57 fmol/mg protein. A similar K-d of 0.2 nM was obtained using isolated membranes from the whole hippocampus, although the number of binding sites was lower (2 fmol/mg protein). Affinity cross-linking of I-125-HU-IL-1 alpha to hippocampal membranes revealed the existence of two types of IL-1 receptor proteins, consistent with the sizes of the type I (85 kD) and type II (60 kD) IL-1 receptor. Oligonucleotide probes were then synthesized and used in RT-PCR followed by Southern blotting to show that the whole brain expresses transcripts for both the type I and type II IL-1 receptors. The murine neuroblastoma cell line, C1300, expresses type I rather than type II IL-1 receptor mRNA. The type I receptor protein can be identified by flow cytometry on the membrane of the C1300 neuronal cell line using indirect immunofluorescence with a rat anti-mouse type I IL-1 receptor MoAb. These data show that mouse brain expresses both type I and type II IL-1 receptor mRNA and proteins and offer further support to the idea that type I IL-1 receptors an synthesized and expressed by neurons.