Cell-permeant NAADP: A novel chemical tool enabling the study of Ca2+ signalling in intact cells

Parkesh, Raman; Lewis, Alexander M.; Aley, Parvinder K.; Arredouani, Abdelilah; Rossi, Sara; Tavares, Ricardo; Vasudevan, Sridhar R.; Rosen, Daniel; Galione, Antony; Dowden, James; Churchill, Grant C.

doi:10.1016/j.ceca.2007.08.006

Cited by 71 publications

(67 citation statements)

References 22 publications

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“…In addition to possible extracellular effects of NAADP [49], we have shown intracellular delivery of NAADP in to cortical neurons and PC12 cells mediates Ca 2+ signals that are inhibited by acid store blockers but readily demonstrable following depletion of endoplasmic reticulum Ca 2+ stores with thapsigargin [50;51]. We also confirmed the presence of NAADP receptors in the former preparation using a novel cell permeable acetoxymethyl ester of NAADP [52]. Importantly, NAADP was shown to potentiate neurite extension in primary neuronal cultures by a mechanism involving IP 3 and ryanodine receptors [50].…”

Section: +supporting

confidence: 59%

Recruitment of NAADP-sensitive acidic Ca2+ stores by glutamate

Pandey

Chuang

Lewis

et al. 2009

Biochemical Journal

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NAADP (nicotinic acid–adenine dinucleotide phosphate) is an unusual second messenger thought to mobilize acidic Ca2+ stores, such as lysosomes or lysosome-like organelles, that are functionally coupled to the ER (endoplasmic reticulum). Although NAADP-sensitive Ca2+ stores have been described in neurons, the physiological cues that recruit them are not known. Here we show that in both hippocampal neurons and glia, extracellular application of glutamate, in the absence of external Ca2+, evoked cytosolic Ca2+ signals that were inhibited by preventing organelle acidification or following osmotic bursting of lysosomes. The sensitivity of both cell types to glutamate correlated well with lysosomal Ca2+ content. However, interfering with acidic compartments was largely without effect on the Ca2+ content of the ER or Ca2+ signals in response to ATP. Glutamate but not ATP elevated cellular NAADP levels. Our results provide evidence for the agonist-specific recruitment of NAADP-sensitive Ca2+ stores by glutamate. This links the actions of NAADP to a major neurotransmitter in the brain.

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Section: +supporting

confidence: 59%

Recruitment of NAADP-sensitive acidic Ca2+ stores by glutamate

Pandey

Chuang

Lewis

et al. 2009

Biochemical Journal

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“…Differentiation was induced with 2% horse serum. NAADP-AM was synthesized in house (18). Ned-19 was obtained from IBScreen (19).…”

Section: Methodsmentioning

confidence: 99%

“…NAADP-acetoxymethylester (NAADP-AM), a recently developed cell-permeant form of NAADP (18), caused a calcium release in these cells and stimulated early and late events of terminal differentiation. Ned-19, a specific inhibitor of the NAADP signaling pathway (19), prevented skeletal muscle differentiation.…”

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confidence: 99%

Nicotinic acid adenine dinucleotide phosphate regulates skeletal muscle differentiation via action at two-pore channels

Aley

Mikołajczyk

Munz

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Calcium signaling is essential for the differentiation of many cell types, including skeletal muscle cells, but its mechanisms remain elusive. Here we demonstrate a crucial role for nicotinic acid adenine dinucleotide phosphate (NAADP) signaling in skeletal muscle differentiation. Although the inositol trisphosphate pathway may have a partial role to play in this process, the ryanodine signaling cascade is not involved. In both skeletal muscle precursors and C2C12, cells interfering with NAADP signaling prevented differentiation, whereas promoting NAADP signaling potentiated differentiation. Moreover, siRNA knockdown of two-pore channels, the target of NAADP, attenuated differentiation. The data presented here strongly suggest that in myoblasts, NAADP acts at acidic organelles on the recently discovered two-pore channels to promote differentiation.

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“…To meet the need for selective and useful chemical probes for studying NAADP signaling, we have synthesized a cell-permeant form of NAADP, NAADP-acetoxymethyl ester, which is cleaved by esterases intracellularly to become biologically active (28), and identified a drug-like, cell-permeant, and selective NAADP antagonist, Ned-19 (see Fig. 1A) (29).…”

Section: Camentioning

confidence: 99%

Analogues of the Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) Antagonist Ned-19 Indicate Two Binding Sites on the NAADP Receptor

Rosen

Lewis

Mizote

et al. 2009

Journal of Biological Chemistry

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Nicotinic acid adenine dinucleotide phosphate (NAADP) is a Ca2؉ -releasing messenger. Biological data suggest that its receptor has two binding sites: one high-affinity locking site and one low-affinity opening site. To directly address the presence and function of these putative binding sites, we synthesized and tested analogues of the NAADP antagonist Ned-19. Ned-19 itself inhibits both NAADP-mediated Ca 2؉ release and NAADP binding. A fluorometry bioassay was used to assess NAADPmediated Ca 2؉ release, whereas a radioreceptor assay was used to assess binding to the NAADP receptor (only at the high-affinity site). In Ned-20, the fluorine is para rather than ortho as in Ned-19. Ned-20 does not inhibit NAADP-mediated Ca 2؉ release but inhibits NAADP binding. Conversely, Ned-19.4 (a methyl ester of Ned-19) inhibits NAADP-mediated Ca 2؉ release but cannot inhibit NAADP binding. Furthermore, Ned-20 prevents the self-desensitization response characteristic of NAADP in sea urchin eggs, confirming that this response is mediated by a high-affinity allosteric site to which NAADP binds in the radioreceptor assay. Collectively, these data provide the first direct evidence for two binding sites (one high-and one lowaffinity) on the NAADP receptor. Ca2ϩ is an extremely versatile and powerful second messenger (1). Three accepted Ca 2ϩ -releasing second messengers that cells utilize to control the spatiotemporal properties of these Ca 2ϩ signals are inositol 1,4,5-trisphosphate (2), cyclic ADPribose (3), and nicotinic acid adenine dinucleotide phosphate (NAADP) 2 (4). Of these three messengers, little is known about the mechanism of action of NAADP. NAADP is the most potent of the three Ca 2ϩ -releasing messengers (5), and its actions are pharmacologically distinct from inositol 1,4,5-trisphosphate and cyclic ADP-ribose (6 -9). Controversy surrounds both the identity of the NAADP-activated channel and its organellar location. There is evidence for the NAADP receptor being the transient receptor potential mucolipin 1 channel (10), the ryanodine receptor (11, 12), or the two-pore channel (Refs. 13-15; for review, see Ref. 16). There is evidence for the Ca 2ϩ store targeted by NAADP being the endoplasmic reticulum (11) or physically distinct (17), acidic, lysosome-related organelles (18).One of the most intriguing features of NAADP is the relationship between concentration, binding, and response (5,9,19). In sea urchin eggs, a subthreshold concentration of NAADP that does not itself release Ca 2ϩ is able to prevent Ca 2ϩ release in response to a second addition of NAADP that would normally be able to release Ca 2ϩ (20,21). This unusual desensitization may relate to the formation of a spatial and temporal "memory" (22,23). This effect is thought to be due to a second high-affinity inhibitory site present on the receptor (24); thus, at low concentrations, NAADP only binds to this site and is able to prevent Ca 2ϩ release via this receptor. In contrast to sea urchin eggs, in mammalian tissue, NAADP has a bell-shaped concentration-r...

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Cell-permeant NAADP: A novel chemical tool enabling the study of Ca2+ signalling in intact cells

Cited by 71 publications

References 22 publications

Recruitment of NAADP-sensitive acidic Ca2+ stores by glutamate

Recruitment of NAADP-sensitive acidic Ca2+ stores by glutamate

Nicotinic acid adenine dinucleotide phosphate regulates skeletal muscle differentiation via action at two-pore channels

Analogues of the Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) Antagonist Ned-19 Indicate Two Binding Sites on the NAADP Receptor

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