Abstract
The molecular mechanisms that mediate and regulate calcium (Ca2+) fluxes through the membranes of intracellular organelles play a key role in the generation and sha** of the local and global cytosolic Ca2+ signals triggering the process of regulated exocytosis in chromaffin cells. Beyond that role, intraorganellar Ca2+ homeostasis also regulates organelle-specific processes such as oxidative phosphorylation in mitochondria, maturation of secretory granules, or stress in the endoplasmic reticulum. In this chapter, we describe current methods to study mitochondrial, endoplasmic reticulum, and secretory vesicle calcium homeostasis in living chromaffin cells using engineered targeted aequorins.
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References
Sparagna GC, Gunter KK, Sheu S-S et al (1995) Mitochondrial calcium uptake from physiological-type pulses of calcium. J Biol Chem 270:27510–27515
Zhao Y, Araki S, Wu J et al (2011) An expanded palette of genetically encoded Ca2+ indicators. Science 333:1888–1891
Suzuki J, Kanemaru K, Iino M (2016) Genetically encoded fluorescent indicators for organellar calcium imaging. Biophys J 111:1119–1131
Simpson USA, Brini AWM, Pozzan M et al (1995) Transfected aequorin in the measurement of cytosolic Ca 2 + concentration ([Ca 2 +]c). J Biol Chem 270:9896–9903
García AG, García-De-Diego AM, Gandía L et al (2006) Calcium signaling and exocytosis in adrenal chromaffin cells. Physiol Rev 86:1093–1131
Montero M, Alonso MT, Carnicero E et al (2000) Chromaffin-cell stimulation triggers fast millimolar mitochondrial Ca2+ transients that modulate secretion. Nat Cell Biol 2:57–61
Rigual R, Montero M, Rico AJ et al (2002) Modulation of secretion by the endoplasmic reticulum in mouse chromaffin cells. Eur J Neurosci 16:1690–1696
Alonso MT, Barrero MJ, Carnicero E et al (1999) Functional measurements of Ca2+ in the endoplasmic reticulum using a herpes virus to deliver targeted aequorin. Cell Calcium 24:87–96
Alonso MT, Barrero MJ, Michelena P et al (1999) Ca2+-induced Ca2+ release in chromaffin cells seen from inside the ER with targeted aequorin. J Cell Biol 144:241–254
Santo-Domingo J, Fonteriz RI, Lobatón CD et al (2010) Ca2+ dynamics in the secretory vesicles of neurosecretory PC12 and INS1 cells. Cell Mol Neurobiol 30:1197–1199
Santo-Domingo J, Vay L, Camacho M et al (2008) Calcium dynamics in bovine adrenal medulla chromaffin cell secretory granules. Eur J Neurosci 28:1265–1274
Inouye S, Noguchi M, Sakaki Y et al (1985) Cloning and sequence analysis of cDNA for the luminescent protein aequorin. Proc Natl Acad Sci USA 82:3154–3158
Cobbold PH, Lee JAC (1991) Aequorin measurements of cytoplasmic free calcium. In: Cellular calcium: a práctical approach. Oxford Press
Li X, Drakulich DA, Zhang P et al (2002) Transduction of bovine adrenal chromaffin cells using a recombinant adenovirus expressing GFP. J Neurosci Methods 122:91–96
Moro M, López M, Gandía L et al (1990) Separation and culture of living adrenaline- and noradrenaline-containing cells from bovine adrenal medullae. Anal Biochem 185:234–238
Montero M, Brini M, Marsault R et al (1995) Monitoring dynamic changes in free Ca2+ concentration in the endoplasmic reticulum of intact cells. EMBO J 14:5467–5475
De la Fuente S, Fonteriz RI, Montero M et al (2013) Ca2+ homeostasis in the endoplasmic reticulum measured with a new low-Ca2+−affinity targeted aequorin. Cell Calcium 54:37–45
De la Fuente S, Fonteriz RI, de La Cruz PJ et al (2012) Mitochondrial free [Ca2+] dynamics measured with a novel low-Ca2+ − affinity aequorin probe. Biochem J 445:371–376
Allen DG, Blinks JR, Prendergast FG (1977) Aequorin luminescence: relation of light emission to calcium concentration-a calcium-independent component. Science 195:996–998
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Santo-Domingo, J., Álvarez-Illera, P., Montenegro, P., Fonteriz, R.I., Montero, M., Álvarez, J. (2023). Measurements of Calcium in Chromaffin Cell Organelles Using Targeted Aequorins. In: Borges, R. (eds) Chromaffin Cells. Methods in Molecular Biology, vol 2565. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2671-9_11
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DOI: https://doi.org/10.1007/978-1-0716-2671-9_11
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