Ca2+ Measurement with Luminescent Probes in the Endoplasmic Reticulum

  1. Alvarez, Javier
  2. Montero, Mayte
Libro:
Measuring Calcium and Calmodulin Inside and Outside Cells

ISBN: 9783642477379,  9783642568510

Año de publicación: 2001

Páginas: 147-163

Tipo: Capítulo de Libro

DOI: 10.1007/978-3-642-56851-0_7 GOOGLE SCHOLAR lock_openAcceso abierto editor

Resumen

The photoprotein aequorin can be used to measure specifically the [Ca2+] inside the lumen of the endoplasmic reticulum ([Ca2+ ER) of intact cells. The main advantage of this technique is the highly precise targeting of the probe. The main disadvantage relies in the relatively low amount of emitted light, which makes it difficult to perform single-cell imaging studies at the required time resolution. On the technical side, the equipment required for luminescence measurements in cell populations is simple and low-cost, but expression of ER-targeted aequorin requires previous transfection or infection to introduce the appropriate DNA plasmid, or alternatively the use of stable cell clones. Finally, calibration of the luminescent records to obtain [Ca2+] values is relatively easy, but interpretation of the results requires taking into account that aequorin is being rapidly consumed during the experiment due to the high levels of Ca2+ (hundreds of micromolar) present in the ER.

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