Novel Cell Line Selectively Expressing Neuropeptide Y‐Y2 Receptors

Novel Cell Line Selectively Expressing Neuropeptide Y‐Y2 Receptors
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选择性表达神经肽 Y-Y2 受体的新型细胞系

DOI:
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发表时间:
2003
期刊:
Journal of Receptor and Signal Transduction Research
影响因子:
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通讯作者:
Annette G Beck Sickinger
Annette G Beck Sickinger
中科院分区:
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文献类型:
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作者:
M. Höfliger;Gloria López Castejón;W. Kiess;Annette G Beck Sickinger

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利用氚标记的神经肽Y(NPY),在放射性结合试验中分析了人神经母细胞瘤细胞系SiMa、Kelly、SH-SY 5 Y、CHP-234和MHH-NB-11对神经肽Y(NPY)的识别。对于细胞系CHP-234和MHH-NB-11,观察到[3 H]丙酰基-NPY结合,通过非线性回归饱和分析测定的Kd值分别为0.64 ± 0.07 nM和0.53 ± 0.12 nM。使用亚型选择性NPY类似物[Leu 31,Pro34]-NPY(NPY-Y1,NPY-Y 5)、[Ahx 5 - 24]-NPY(NPY-Y2)、[Ala 31,Aib 32]-NPY(NPY-Y 5)、NPY [3 - 36](NPY-Y2,NPY-Y 5)和NPY [13 - 36](NPY-Y2),通过竞争分析确定受体亚型。两种细胞系CHP-234和MHH-NB-11(后者首次表征NPY受体)均显示出NPY-Y2受体的排他性表达。在两种细胞系中,NPY的结合诱导了信号转导,这在ELISA中被监测为毛喉素诱导的cAMP产生的减少。
Abstract Neuropeptide Y (NPY) recognition by the human neuroblastoma cell lines SiMa, Kelly, SH‐SY5Y, CHP‐234, and MHH‐NB‐11 was analyzed in radioactive binding assays using tritiated NPY. For the cell lines CHP‐234 and MHH‐NB‐11 binding of [3H]propionyl‐NPY was observed with Kd‐values of 0.64 ± 0.07 nM and 0.53 ± 0.12 nM, respectively, determined by saturation analysis with non‐linear regression. The receptor subtype was determined by competition analysis using the subtype selective NPY analogues [Leu31, Pro34]‐NPY (NPY‐Y1, NPY‐Y5), [Ahx5‐24]‐NPY (NPY‐Y2), [Ala31, Aib32]‐NPY (NPY‐Y5), NPY [3‐36] (NPY‐Y2, NPY‐Y5), and NPY [13‐36] (NPY‐Y2). Both cell lines, CHP‐234 and MHH‐NB‐11, the latter one being characterized for NPY receptors for the first time, showed exclusive expression of NPY‐Y2 receptors. In both cell lines binding of NPY induced signal transduction, which was monitored as reduction of forskolin‐induced cAMP production in an ELISA.
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