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β2 肾上腺素能受体β-arrestin2 募集:一种适应于药理学本科培训的荧光素酶互补测定法。

β-arrestin2 recruitment at the β2 adrenergic receptor: A luciferase complementation assay adapted for undergraduate training in pharmacology.

机构信息

Neuropharmacology Laboratory, Institute of Neuroscience, UCLouvain, Brussels, Belgium.

Faculty of Pharmacy and Biomedical Sciences, UCLouvain, Brussels, Belgium.

出版信息

Pharmacol Res Perspect. 2021 Feb;9(1):e00706. doi: 10.1002/prp2.706.

Abstract

In the context of pharmacology teaching, hands-on activities constitute an essential complement to theoretical lectures. Frequently, these activities consist in exposing fresh animal tissues or even living animals to selected drugs and qualitatively or quantitatively evaluating functional responses. However, technological advancements in pharmacological research and the growing concerns for animal experimentation support the need for innovative and flexible in vitro assays adapted for teaching purposes. We herein report the implementation of a luciferase complementation assay (LCA) enabling to dynamically monitor β-arrestin2 recruitment at the β adrenergic receptor in the framework of pharmacological training at the faculty of Pharmacy and Biomedical Sciences. The assay allowed students to quantitatively characterize the competitive antagonism of propranolol, and to calculate pEC , pK , and pA values after a guided data analysis session. Moreover, the newly implemented workshop delivered highly reproducible results and were generally appreciated by students. As such, we report that the luciferase complementation-based assay proved to be a straightforward, robust, and cost-effective alternative to experiments performed on animal tissues, constituting a useful and flexible tool to enhance and update current hands-on training in the context of pharmacological teaching.

摘要

在药理学教学中,实践活动是理论讲座的重要补充。这些活动通常包括将新鲜的动物组织甚至活体动物暴露于选定的药物中,并定性或定量地评估功能反应。然而,药理学研究的技术进步和对动物实验的日益关注支持了创新和灵活的体外检测的需求,这些检测适用于教学目的。我们在此报告了一种荧光素酶互补测定法(LCA)的实施,该方法能够在药学院和生物医学科学系的药理学培训框架内动态监测β肾上腺素能受体的β-arrestin2 募集。该测定法允许学生定量表征普萘洛尔的竞争性拮抗作用,并在经过引导的数据分析会议后计算 pEC 、pK 和 pA 值。此外,新实施的研讨会提供了高度可重复的结果,并受到学生的普遍赞赏。因此,我们报告说,基于荧光素酶互补的测定法是一种简单、稳健且具有成本效益的替代动物组织实验的方法,是增强和更新药理学教学中当前实践培训的有用且灵活的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cff0/7842874/dfbd0510425f/PRP2-9-e00706-g001.jpg

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