| Quantitative Determination of Ca2+-binding to Ca2+-sensor Proteins by Isothermal Titration Calorimetry
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Author:
Date:
2020-04-05
[Abstract] Diverse and complex molecular recognitions are central elements of signal transduction cascades. The strength and nature of these interaction modes can be determined by different experimental approaches. Among those, Isothermal titration calorimetry (ITC) offers certain advantages by providing binding constants and thermodynamic parameters from titration series without a need to label or immobilize one or more interaction partners. Furthermore, second messenger homeostasis involving Ca2+-ions requires in particular knowledge about stoichiometries and affinities of Ca2+-binding to Ca2+-sensor proteins or Ca2+-dependent regulators, which can be obtained by employing ITC. We used ITC to measure these parameters for a set of neuronal Ca2+-sensor ...
[摘要] [摘要] 多样而复杂的分子识别是信号转导级联的核心要素。这些相互作用模式的强度和性质可以通过不同的实验方法来确定。其中，等温滴定热量法（ITC）通过提供来自滴定系列的结合常数和热力学参数，而无需标记或固定一个或多个相互作用伙伴，从而提供了某些优势。此外，涉及钙第二信使动态平衡2+ -离子需要小号特别知识大约化学计量和Ca的亲和力2+ -结合对于Ca 2+ 多个传感器蛋白或钙2+ 依赖的调节器，可以通过使用ITC获得。我们使用ITC来测量在感光细胞中运行的一组神经元Ca 2+ 传感器蛋白的这些参数。在这里，我们提出了一个分步协议，以（a）测量与Ca 2+ 传感器鸟苷酸环化酶e激活蛋白1的Ca 2+ 相互作用，（b）设计ITC实验并准备样品，（c）去除Ca 2+ 几乎完全来自Ca 2+ 结合蛋白，而无需使用螯合剂（如EGTA）。
[背景 ] ...
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| Mouse Subependymal Zone Explants Cultured on Primary Astrocytes
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Author:
Date:
2016-07-20
[Abstract] Neuroblast migration is a major component in the process of neuronal circuit assembly. In the rodent postnatal brain, the subependymal zone (SEZ) is the largest neurogenic niche where new neurons are born throughout life. These cells migrate several millimeters following a specific pathway called the rostral migratory stream (RMS) towards the olfactory bulb. Once they reach their final destination, they differentiate mainly as interneurons and integrate into already established neuronal circuits. Along the RMS, neuroblasts use a particular mode of migration known as chain migration. They stay attached to each other while migrating and are intimately associated with adjacent astrocytic processes. To dissect the molecular and cellular signals involved in neuroblast migration, we developed ...
[摘要] 神经母细胞迁移是神经元电路组装过程中的主要组成部分。 在啮齿动物出生后的大脑中，子宫颈区（SEZ）是新生神经元生命中最大的神经源性生态位。 这些细胞沿着称为传播流（RMS）的特定途径朝向嗅球迁移几毫米。 一旦达到其最终目的地，它们主要区分为中间神经元并整合到已建立的神经元回路中。 沿着RMS，神经母细胞使用称为链转移的特定迁移模式。 它们在迁移时保持相互依赖，并与相邻的星形细胞过程密切相关。 为了解剖神经母细胞迁移中涉及的分子和细胞信号，我们开发了一种体外系统，其由在星形胶质细胞单层顶部共培养的经SEZ衍生的外植体组成。
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