Identification of functional receptors for ciliary neurotrophic factor on chick ciliary ganglion neurons.
Identification of functional receptors for ciliary neurotrophic factor on chick ciliary ganglion neurons.
复制标题
鉴定鸡睫状神经节神经元上睫状神经营养因子的功能受体。
DOI：
10.1016/0306-4522(95)00560-9
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发表时间：
1996
期刊：
影响因子：
3.3
通讯作者：
Halvorsen,SW
中科院分区：
文献类型：
--
作者：
Koshlukova,S;Finn,TP;Nishi,R;Halvorsen,SW
睫状神经营养因子和禽类同源物，生长促进活性，是营养因子的细胞因子/神经因子家族的成员，并且已经被提出作为睫状神经节神经元在体内的存活和发育因子起作用。在这里，我们确定了第一次睫状神经营养因子和生长促进活性的功能受体培养睫状神经节神经元。[125 I]大鼠睫状神经营养因子结合研究表明，大鼠睫状神经营养因子和生长促进活性以单一亲和力结合这些受体，而人睫状神经营养因子识别高亲和力和低亲和力位点。在生物测定中，人睫状神经营养因子和禽类生长促进活性的相对效力的比较表明，生长促进活性在促进存活和调节乙酰胆碱受体方面的活性高3至5倍。睫状神经营养因子的结合是特异性的，对磷脂酰肌醇特异性磷脂酶C敏感，并被白血病抑制因子部分抑制，但不被人类神经因子家族的其他成员抑制，包括白细胞介素-6、白细胞介素-11和制瘤素M。[125 I]大鼠睫状神经营养因子与睫状神经元的交联导致估计分子量为153，000、81，000和72，000的三种蛋白质的特异性标记。在用磷脂酰肌醇特异性磷脂酶C处理后，仅81，000分子量组分从细胞中释放，表明通过糖基磷脂酰肌醇连接的膜附着。用睫状神经营养因子或生长促进活性刺激，而不是用其他神经因子刺激，导致90，000分子量蛋白的快速酪氨酸磷酸化，其被磷脂酰肌醇特异性磷脂酶C预处理抑制。总之，我们在这里报告的药理学和功能特性的睫状神经营养因子受体对胚胎睫状神经节神经元。这些结果为阐明睫状神经营养因子作用的分子机制和理解其在特定神经元群体中的生理作用提供了手段。
Ciliary neurotrophic factor and an avian homolog, growth promoting activity, are members of the cytokine/neurokine family of trophic factors and have been proposed to function as survival and developmental factors for ciliary ganglion neurons in vivo. Here we identify for the first time functional receptors for ciliary neurotrophic factor and growth promoting activity on cultured ciliary ganglion neurons. [125I]Rat ciliary neurotrophic factor binding studies indicate that rat ciliary neurotrophic factor and growth promoting activity bind to these receptors with a single affinity, while human ciliary neurotrophic factor recognizes both a high- and low-affinity site. Comparison of the relative potency of human ciliary neurotrophic factor and avian growth promoting activity in biological assays indicates that growth promoting activity is three to five times more active in promoting survival and in regulating acetylcholine receptors. The binding of ciliary neurotrophic factor is specific, sensitive to phosphatidylinositol-specific phospholipase C and partially inhibited by leukemia inhibitory factor, but not inhibited by other members of the human neurokine family, including interleukin-6, interleukin-11 and oncostatin M. Cross-linking of [125I]rat ciliary neurotrophic factor to ciliary neurons results in the specific labeling of three proteins with estimated molecular masses of 153,000, 81,000 and 72,000. Only the 81,000 molecular weight component is released from the cells after treatment with phosphatidylinositol-specific phospholipase C, suggesting a membrane attachment via a glycosylphosphatidylinositol linkage. Stimulation with ciliary neurotrophic factor or growth promoting activity, but not by other neurokines, results in the rapid tyrosine phosphorylation of a 90,000 molecular weight protein that is inhibited by pretreatment with phosphatidylinositol-specific phospholipase C. In conclusion, we report here the pharmacological and functional properties of ciliary neurotrophic factor receptors on embryonic ciliary ganglion neurons. These results provide the means for elaborating the molecular mechanisms of ciliary neurotrophic factor action and understanding its physiological role in a defined neuronal population.