Titlebook: Calcium Signaling; Methods and Protocol Caroline M. Gorvin Book 2025 The Editor(s) (if applicable) and The Author(s), under exclusive licen

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https://doi.org/10.1007/978-94-015-8348-0y and/or calcium-dependent subcellular signaling. Here, we describe a protocol for repeated two-photon imaging of calcium signals in mice expressing a genetically encoded calcium indicator that have been implanted with a chronic cranial window.

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A Translational 3-dof Parallel Manipulatora.). Using mice expressing a genetically encoded Ca. indicator (GCaMP6f), we present in this chapter a method to visualize at high spatiotemporal resolution changes in intracellular Ca. in mammary epithelial cells, both in vitro (2D) and ex vivo (3D). The procedure to optimally prepare mammary tissue and primary cells is presented in detail.

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Measuring IP3 Generation in Real-Time Using a NanoBiT Luminescence Biosensorssure regulation, and calcium homeostasis. Activation of Gq/11-coupled receptors stimulates the generation of inositol 1,4,5-trisphosphate (IP3), which mobilizes intracellular calcium release from the endoplasmic reticulum. This chapter describes an assay that uses a NanoBiT-IP3 luminescent biosenso

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Monitoring Calcium-Sensing Receptor (CaSR)-Induced Intracellular Calcium Flux Using an Indo-1 Flow Cns in the receptor, or components of its signaling and trafficking pathway, cause disorders of calcium homeostasis. Inactivating mutations cause neonatal severe hyperparathyroidism or familial hypocalciuric hypercalcemia (FHH), while gain-of-function mutations cause autosomal dominant hypocalcemia (

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Bioluminescence Resonance Energy Transfer (BRET) Assay to Measure Gq Recruitment to the Ghrelin ReceBRET). Focusing on the eBRET. and NanoBRET™ variants, the chapter covers steps from cell culture to transfection, ligand stimulation, and BRET measurements, offering a robust protocol to examine the temporal aspects of Gq signaling in live cells. This methodology facilitates a nuanced understanding

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Analysis of Calcium-Sensing Receptor Signaling Using Dual Luciferase Assays), a G protein-coupled receptor (GPCR), induces characteristic signaling pathways that stimulate extracellular signal-regulated kinase 1/2 (ERK1/2) and Ca. mobilization from the endoplasmic reticulum. ERK1/2 causes an activation of the serum response element (SRE), whereas Ca. causes an activation o