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Monitoring calcium-sensing receptor (CaSR)-induced intracellular calcium flux using an Indo-1 flow cytometry assay
- Abstract:
- The calcium-sensing receptor (CaSR) has a critical role in maintaining serum calcium concentrations within the normal physiological range, and mutations 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 (ADH). Characterizing the functional impact of mutations of the CaSR, and components of the CaSR-signaling pathway, is clinically important to enable correct diagnoses of FHH and ADH, optimize management, and prevent inappropriate parathyroidectomy or vitamin D supplementation. CaSR signals predominantly by activating the G-alpha subunit-11 to mobilize calcium release from intracellular stores. Thus, measurement of CaSR-induced intracellular calcium (Ca2+i) signaling is the gold standard method to investigate the pathogenicity of CaSR genetic variants. This protocol describes a method to assess CaSR-induced Ca2+I signaling using the Indo-1 calcium indicator dye and flow cytometry. This method has been used to assess multiple genetic variants in CaSR and components of its signaling and trafficking pathway in HEK293 cells.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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- Files:
-
-
(Preview, Accepted manuscript, pdf, 227.6KB, Terms of use)
-
- Publisher copy:
- 10.1007/978-1-0716-4164-4_4
Authors
+ Wellcome Trust
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- Funder identifier:
- https://ror.org/029chgv08
- Grant:
- 220668/Z/20/Z
- Publisher:
- Springer
- Host title:
- Calcium Signaling
- Pages:
- 43-55
- Series:
- Methods in Molecular Biology
- Series number:
- 2861
- Place of publication:
- United States
- Publication date:
- 2024-10-13
- DOI:
- EISSN:
-
1940-6029
- ISSN:
-
1064-3745
- Pmid:
-
39395096
- EISBN:
- 9781071641644
- ISBN:
- 9781071641637
- Language:
-
English
- Keywords:
- Pubs id:
-
2045036
- Local pid:
-
pubs:2045036
- Deposit date:
-
2024-10-29
Terms of use
- Copyright holder:
- Gorvin and Howles
- Copyright date:
- 2025
- Rights statement:
- © 2025 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
- Notes:
- This is the accepted manuscript version of the chapter. The final version is available online from Springer at https://dx.doi.org/10.1007/978-1-0716-4164-4_4
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