School of Biological Sciences, University of Auckland, Auckland, New Zealand.
Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland, New Zealand.
Am J Physiol Regul Integr Comp Physiol. 2021 May 1;320(5):R697-R703. doi: 10.1152/ajpregu.00338.2020. Epub 2021 Feb 10.
Amylin is a 37-amino acid endocrine hormone secreted from the pancreas in response to nutrient intake, acting centrally to promote meal-ending satiation. With many studies linking amylin action to the nervous system, determining the distribution or expression of amylin in the nervous system is critical. However, amylin shares sequence identity and structural homology to the related neuropeptide calcitonin gene-related peptide (CGRP). This creates challenges in identifying selective amylin antibodies that do not cross-react with CGRP, especially in neural tissues, where CGRP is densely packed into secretory vesicles. Here, we characterized eight amylin antibodies to determine their ability to detect amylin and cross-react with rat or human αCGRP, using immunoblots and preabsorption controls in rat pancreas. We observed that amylin antibodies frequently cross-reacted with αCGRP and are therefore not suitable for use in tissues that highly express CGRP. Earlier work using these antibodies should be revisited in light of our findings.
胰岛淀粉样多肽是一种 37 个氨基酸的内分泌激素，在进食后由胰腺分泌，作用于中枢以促进进食结束时的饱腹感。由于许多研究将胰岛淀粉样多肽的作用与神经系统联系起来，因此确定其在神经系统中的分布或表达至关重要。然而，胰岛淀粉样多肽与相关神经肽降钙素基因相关肽 (CGRP) 具有序列同一性和结构同源性。这在识别不与 CGRP 发生交叉反应的选择性胰岛淀粉样多肽抗体时带来了挑战，尤其是在神经组织中，CGRP 被密集地包装到分泌小泡中。在这里，我们对八种胰岛淀粉样多肽抗体进行了表征，以确定它们在使用免疫印迹和大鼠胰腺的预吸收对照检测大鼠或人αCGRP 时检测胰岛淀粉样多肽的能力和交叉反应性。我们观察到胰岛淀粉样多肽抗体经常与αCGRP 发生交叉反应，因此不适合用于高度表达 CGRP 的组织。鉴于我们的发现，应该重新审视使用这些抗体的早期工作。
