Ectonucleotide pyrophosphatase/phosphodiesterase activity in Neuro‐2a neuroblastoma cells: changes in expression associated with neuronal differentiation

Gómez‐Villafuertes, Rosa; Pintor, Jesús; Miras-Portugal, Marı́a Teresa; Gualix, Javier

doi:10.1111/jnc.12794

Cited by 15 publications

(19 citation statements)

References 40 publications

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“…This tight regulation mainly focuses on controlling the release and the removal of nucleotides from the extracellular space. In terms of extracellular nucleotide removal, these compounds can be rapidly hydrolyzed to the last product adenosine, subsequently transported and incorporated to the cellular nucleotide pool ( Rotllan and Miras Portugal, 1985 ; Miras-Portugal et al, 1986 ; Sen et al, 1998 ; Zimmermann et al, 2012 ; Gomez-Villafuertes et al, 2014 ; Pastor-Anglada and Perez-Torras, 2015 ).…”

Section: Discussionmentioning

confidence: 99%

Specific Temporal Distribution and Subcellular Localization of a Functional Vesicular Nucleotide Transporter (VNUT) in Cerebellar Granule Neurons

et al. 2017

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Adenosine triphosphate (ATP) is an important extracellular neurotransmitter that participates in several critical processes like cell differentiation, neuroprotection or axon guidance. Prior to its exocytosis, ATP must be stored in secretory vesicles, a process that is mediated by the Vesicular Nucleotide Transporter (VNUT). This transporter has been identified as the product of the SLC17A9 gene and it is prominently expressed in discrete brain areas, including the cerebellum. The main population of cerebellar neurons, the glutamatergic granule neurons, depends on purinergic signaling to trigger neuroprotective responses. However, while nucleotide receptors like P2X7 and P2Y13 are known to be involved in neuroprotection, the mechanisms that regulate ATP release in relation to such events are less clearly understood. In this work, we demonstrate that cerebellar granule cells express a functional VNUT that is involved in the regulation of ATP exocytosis. Numerous vesicles loaded with this nucleotide can be detected in these granule cells and are staining by the fluorescent ATP-marker, quinacrine. High potassium stimulation reduces quinacrine fluorescence in granule cells, indicating they release ATP via calcium dependent exocytosis. Specific subcellular markers were used to assess the localization of VNUT in granule cells, and the transporter was detected in both the axonal and somatodendritic compartments, most predominantly in the latter. However, co-localization with the specific lysosomal marker LAMP-1 indicated that VNUT can also be found in non-synaptic vesicles, such as lysosomes. Interestingly, the weak co-localization between VNUT and VGLUT1 suggests that the ATP and glutamate vesicle pools are segregated, as also observed in the cerebellar cortex. During post-natal cerebellar development, VNUT is found in granule cell precursors, co-localizing with markers of immature cells like doublecortin, suggesting that this transporter may be implicated in the initial stages of granule cell development.

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Section: Discussionmentioning

confidence: 99%

Specific Temporal Distribution and Subcellular Localization of a Functional Vesicular Nucleotide Transporter (VNUT) in Cerebellar Granule Neurons

et al. 2017

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“…Increase in NPP1 expression was found to correlate with the aggressiveness of astrocytic brain tumors (Aerts et al, 2011 ). NPP1 has also been reported to be expressed in N2a mouse neuroblastoma cells, and its expression level was reduced when the cells differentiated into a neuronal-like phenotype (Gómez-Villafuertes et al, 2014 ). Thus, NPP1 inhibitors might be useful for the treatment of brain cancers.…”

Section: Introductionmentioning

confidence: 99%

Substrate-Dependence of Competitive Nucleotide Pyrophosphatase/Phosphodiesterase1 (NPP1) Inhibitors

et al. 2017

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Nucleotide pyrophosphatase/phosphodiesterase type 1 (NPP1) is a membrane glycoprotein involved in the hydrolysis of extracellular nucleotides. Its major substrate is ATP which is converted to AMP and diphosphate. NPP1 was proposed as a new therapeutic target in brain cancer and immuno-oncology. Several NPP1 inhibitors have been reported to date, most of which were evaluated vs. the artificial substrate p-nitrophenyl 5 ′ -thymidine monophosphate (p-Nph-5 ′ -TMP). Recently, we observed large discrepancies in inhibitory potencies for a class of competitive NPP1 inhibitors when tested vs. the artificial substrate p-Nph-5 ′ -TMP as compared to the natural substrate ATP. Therefore, the goal of the present study was to investigate whether inhibitors of human NPP1 generally display substrate-dependent inhibitory potency. Systematic evaluation of nucleotidic as well as non-nucleotidic NPP1 inhibitors revealed significant differences in determined K i values for competitive, but not for non-and un-competitive inhibitors when tested vs. the frequently used artificial substrate p-Nph-5 ′ -TMP as compared to ATP. Allosteric modulation of NPP1 by p-Nph-5 ′ -TMP may explain these discrepancies. Results obtained using the AMP derivative p-nitrophenyl 5 ′ -adenosine monophosphate (p-Nph-5 ′ -AMP) as an alternative artificial substrate correlated much better with those employing the natural substrate ATP.

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“…At very low level, NPP1 is also expressed on epithelium of salivary gland ducts and brain capillary endothelium . Another isozyme of NPPs, h ‐NPP3 is highly expressed in hepatocytes, small intestine, uterus and prostate …”

Section: Introductionmentioning

confidence: 99%

“…[6] Another isozyme of NPPs, h-NPP3 is highly expressed in hepatocytes, small intestine, uterus and prostate. [7] At a physiological level, NPPs mainly affect a number of processes like bone mineralization, cell proliferation, signaling by insulin and nucleotide, digestion and motility. [8] NPP1 plays an significant role in bone mineralization as it acts as the important enzyme responsible for the production of PPi for osteoblast and chondrocytes.…”

Section: Introductionmentioning

confidence: 99%

Benzo[b]carbazolediones Synthesis and Inhibitory Effects on Nucleotide Pyrophosphatases/Phosphodiesterases

Ejaz

Molenda

et al. 2019

ChemistrySelect

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Two new and efficient methods for the synthesis of benzo[b]carbazolediones have been developed. Various derivatives were synthesized by either a three‐component one‐pot reaction or a two‐component domino reaction in moderate to good yield. Moreover, inhibition studies of these compounds against nucleotide pyrophosphatases (NPPs) have been carried out. An interesting and promising inhibitory effect was observed by the influence of different substituents. Substitution with a methyl group located at the indole moiety was found sensitive towards h‐NPP1 (4b; IC50 ±SEM = 0.57 ± 0.05 μM), while the unsubstituted derivative exhibited a higher sensitivity towards h‐NPP3 (4a; IC50 ± SEM = 0.16 ± 0.06 μM). Both derivatives presented non‐selective inhibition of both isozymes. Among all the derivatives, two derivatives with anisyl groups showed the highest selectivity towards h‐NPP3 and no interaction with h‐NPP1. Finally, the free binding energies were calculated and molecular docking studies were performed in order to provide an insight into putative binding modes of these inhibitors.

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Ectonucleotide pyrophosphatase/phosphodiesterase activity in Neuro‐2a neuroblastoma cells: changes in expression associated with neuronal differentiation

Cited by 15 publications

References 40 publications

Specific Temporal Distribution and Subcellular Localization of a Functional Vesicular Nucleotide Transporter (VNUT) in Cerebellar Granule Neurons

Specific Temporal Distribution and Subcellular Localization of a Functional Vesicular Nucleotide Transporter (VNUT) in Cerebellar Granule Neurons

Substrate-Dependence of Competitive Nucleotide Pyrophosphatase/Phosphodiesterase1 (NPP1) Inhibitors

Benzo[b]carbazolediones Synthesis and Inhibitory Effects on Nucleotide Pyrophosphatases/Phosphodiesterases

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