Pei G, Ling K, Pu L, Cunningham M D, Ma L
Shanghai Institute of Cell Biology, Chinese Academy of Sciences, People's Republic of China.
FEBS Lett. 1997 Jul 21;412(1):253-6. doi: 10.1016/s0014-5793(97)00790-4.
A Chinese hamster ovary (CHO) cell line, CHO-ORL1, stably expressing human opioid receptor-like receptor 1 (ORL1) has been used to determine ORL1-mediated signaling events using microphysiometry. Nociceptin/orphanin FQ (N/OFQ), a specific endogenous agonist of ORL1, induced an increase in extracellular acidification rate (ECAR) in CHO-ORL1 cells. The ECAR response stimulated by N/OFQ was concentration-dependent and pertussis toxin-sensitive. Repeated exposures of the cells to N/OFQ caused desensitization of ORL1. The ECAR response was recovered at the half-life of approximately 12 min after the initial challenge. Pretreatment with inhibitor of cAMP-dependent kinase did not affect desensitization of ORL1. However, specific inhibitors for protein kinase C almost abolished N/OFQ-induced desensitization of extracellular acidification responsiveness, indicating the involvement of protein kinase C in the process.
一种稳定表达人阿片样受体1（ORL1）的中国仓鼠卵巢（CHO）细胞系CHO-ORL1，已被用于通过微流控技术确定ORL1介导的信号转导事件。痛敏肽/孤啡肽（N/OFQ）是ORL1的一种特异性内源性激动剂，可诱导CHO-ORL1细胞的细胞外酸化率（ECAR）增加。N/OFQ刺激的ECAR反应具有浓度依赖性且对百日咳毒素敏感。细胞反复暴露于N/OFQ会导致ORL1脱敏。在初始刺激后约12分钟的半衰期时，ECAR反应恢复。用环磷酸腺苷依赖性激酶抑制剂预处理不影响ORL1的脱敏。然而，蛋白激酶C的特异性抑制剂几乎完全消除了N/OFQ诱导的细胞外酸化反应性脱敏，表明蛋白激酶C参与了这一过程。
