Department of Clinical Genetics, UMC Amsterdam, the Netherlands.
Department of Functional Genomics, Center for Neurogenomics and Cognitive Research (CNCR), Vrije Universiteit (VU) Amsterdam, de Boelelaan 1085, 1081 HV Amsterdam, the Netherlands.
STAR Protoc. 2021 Feb 3;2(1):100325. doi: 10.1016/j.xpro.2021.100325. eCollection 2021 Mar 19.
Neuropeptides are essential signaling molecules secreted by dense-core vesicles (DCVs). They contribute to information processing in the brain, controlling a variety of physiological conditions. Defective neuropeptide signaling is implicated in several psychiatric disorders. Here, we provide a protocol for the quantitative analysis of DCV fusion events in rodent neurons using pH-sensitive DCV fusion probes and custom-written analysis algorithms. This method can be used to study DCV fusion mechanisms and is easily adapted to investigate fusion principles of other secretory organelles. For complete details on the use and execution of this protocol, please refer to Persoon et al. (2019).
神经肽是由致密核心囊泡(DCV)分泌的重要信号分子。它们有助于大脑中的信息处理,控制各种生理状况。神经肽信号传递缺陷与多种精神疾病有关。在这里,我们提供了一种使用 pH 敏感的 DCV 融合探针和自定义分析算法定量分析啮齿动物神经元中 DCV 融合事件的方案。该方法可用于研究 DCV 融合机制,并且易于适应研究其他分泌细胞器的融合原理。有关该方案使用和执行的完整详细信息,请参阅 Persoon 等人。(2019 年)。