Radioligand Binding to Quantify Adrenergic Receptor Expression in the Heart.
Radioligand Binding to Quantify Adrenergic Receptor Expression in the Heart.
复制标题
放射性配体结合量化心脏中肾上腺素受体的表达。
DOI：
10.1002/cpz1.649
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Grisanti,LaurelA
中科院分区：
文献类型：
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作者：
Grisanti,LaurelA
β-肾上腺素能受体调节健康和衰竭心脏的心脏功能。在心力衰竭中，由于交感神经系统的慢性过度激活，它们的表达减少，导致心功能下降和疾病进展。此外，防止β-肾上腺素能受体下调或恢复β-肾上腺素能受体水平的治疗是有益的，使心脏β-肾上腺素能受体表达的测定成为一个重要的考虑因素。虽然定量逆转录聚合酶链式反应可以提供β-肾上腺素能受体密度和亚型表达的指示，但基因水平并不总是与功能蛋白水平相关。此外，针对β-肾上腺素能受体的抗体缺乏特异性，使得免疫印迹和其他基于抗体的技术不可靠。放射配基结合分析是在50多年前发展起来的，至今仍是定量生物样品中β-肾上腺素能受体密度的金标准。这项技术利用了高亲和力、高特异性的配体与受体的结合，可以给出受体表达的可量化水平。此外，使用亚型选择性拮抗剂的竞争分析产生结合图谱，并可以区分心肌组织中β-肾上腺素能受体亚型的表达。本文利用饱和和竞争放射配基结合技术对心脏β受体进行定量，以定量心肌细胞膜上的β受体密度和配体亲和力。2023 Wiley期刊LLC基本方案：放射配基结合以量化心脏中肾上腺素能受体的表达
β‐adrenergic receptors regulate cardiac function in both the healthy and failing heart. Their expression is decreased in heart failure due to chronic overactivation of the sympathetic nervous system, contributing to declines in cardiac function and disease progression. Furthermore, therapies that prevent β‐adrenergic receptor downregulation or restore β‐adrenergic receptor levels are beneficial, making the determination of cardiac β‐adrenergic receptor expression in the heart an important consideration. Although quantitative RT‐PCR can provide an indication of β‐adrenergic receptor density and subtype expression, mRNA levels do not always correlate with functional protein levels. Additionally, antibodies to β‐adrenergic receptors lack specificity, making immunoblotting and other antibody‐based techniques unreliable. Radioligand binding assays were developed over 50 years ago and remain the gold standard for quantifying β‐adrenergic receptor densities in biological samples. This technique capitalizes on the binding of high‐affinity, highly specific ligands to receptors and can give quantifiable levels of receptor expression. Furthermore, competition assays using subtype‐selective antagonists generate binding profiles and can differentiate β‐adrenergic receptor subtype expression in cardiac tissue. This article focuses on the quantification of β‐adrenergic receptors in the heart using saturation and competition radioligand binding techniques to quantify β‐adrenergic receptor density and ligand affinities in cardiac membranes. © 2023 Wiley Periodicals LLC.Basic Protocol: Radioligand binding to quantify adrenergic receptor expression in the heart