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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Calcium Imaging in Brain Tissue Slicescium ions in tissues. This widely used method has provided significant advances to our understanding of cellular signal transduction, including the discovery of subcellular compartmentalization of neurons and astrocytes, the identification of multiple signaling pathways, and mapping the functional c

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Two-Photon Microscopy to Measure Calcium Signaling in the Living Brainhanges in the brightness of fluorescent calcium indicators provide a readout of calcium levels over time, affording information about neuronal activity and/or calcium-dependent subcellular signaling. Here, we describe a protocol for repeated two-photon imaging of calcium signals in mice expressing a

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Measuring Calcium-Sensing Receptor Agonist-Driven Insertional Signaling (ADIS) and Trafficking by TIicking mechanisms to respond to constant exposure to its orthosteric ligand calcium. CaSR rapidly responds to fluctuations in serum calcium by driving forward trafficking of the receptor to cell surfaces in a mechanism known as agonist-driven insertional signaling (ADIS). This increase in CaSR at ce

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Measuring Calcium Signaling at the Primary Ciliacritical role in cellular functions, the intricate machinery of molecular signaling has been intensively studied. A diverse arsenal of techniques exists to quantify the molecules involved in these processes. Among them, calcium stands out as a ubiquitous signaling molecule with roles in countless bi

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Monitoring ER Ca2+ by Luminescence with Low Affinity GFP-Aequorin Protein (GAP)release it upon cell activation. Monitoring of Ca. dynamics within the ER lumen is best achieved using genetically encoded and targeted reporters. Luminescent probes based on the photoprotein aequorin have provided significant insight to measure subcellular Ca.. Here we describe a robust and quantit

DEMUR

Analyzing Mitochondrial Calcium Influx in Isolated Mitochondriaety of pathways including energy production and cell death. Dysregulation of mitochondrial Ca. homeostasis promotes pathologies including neurodegenerative diseases, cardiovascular disorders, and metabolic syndromes. The significance of mitochondria to Ca. signaling and regulation underscores the va

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Methods for Imaging Intracellular Calcium Signals in the Mouse Mammary Epithelium in Two and Three Din the functionally mature mammary gland in response to oxytocin are essential for milk ejection and are tightly regulated by intracellular calcium (Ca.). Using mice expressing a genetically encoded Ca. indicator (GCaMP6f), we present in this chapter a method to visualize at high spatiotemporal reso

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Monitoring Calcium-Sensing Receptor (CaSR)-Induced Intracellular Calcium Flux Using an Indo-1 Flow Cum (Ca.) signaling is the gold standard method to investigate the pathogenicity of CaSR genetic variants. This protocol describes a method to assess CaSR-induced Ca. signaling using the Indo-1 calcium indicator dye and flow cytometry. This method has been used to assess multiple genetic variants in