High Sensitivity, Quantitative Measurements of Polyphosphate Using a New DAPI-Based Approach

Aschar‐Sobbi, Roozbeh; Abramov, Andrey Y.; Diao, Catherine; Kargacin, Margaret E.; Kargacin, Gary J.; French, Robert J.; Pavlov, Evgeny

doi:10.1007/s10895-008-0315-4

Cited by 213 publications

(217 citation statements)

References 29 publications

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“…Emission fluorescence of DAPI shifts to longer wavelengths in the presence of polyP: this change is specific for polyP and is not produced by PPi, DNA, or other anions. 23,25 In U266 MC, we found differential fluorescent labeling with DAPI when settings for DNA or polyP detection were used (Online Supplementary Figure S1A). We have also found different patterns of labeling using a specific method for RNA detection (Online Supplementary Figure S1B).…”

Section: Polyphosphate Is Present In Myeloma Cell Linesmentioning

confidence: 98%

“…The method of polyP detection using DAPI was first described in the early 1980s. 21 Since that time, this procedure has been used in several studies (some examples are references: 6,8,13,16,[22][23][24][25] ). Emission fluorescence of DAPI shifts to longer wavelengths in the presence of polyP: this change is specific for polyP and is not produced by PPi, DNA, or other anions.…”

Section: Polyphosphate Is Present In Myeloma Cell Linesmentioning

confidence: 99%

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Myeloma cells contain high levels of inorganic polyphosphate which is associated with nucleolar transcription

Jimenez-Nuñez¹,

Moreno-Sanchez²,

Hernández-Ruiz³

et al. 2012

Haematologica

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The online version of this article has a Supplementary Appendix. BackgroundIn hematology there has recently been increasing interest in inorganic polyphosphate. This polymer accumulates in platelet granules and its functions include modulating various stages of blood coagulation, inducing angiogenesis, and provoking apoptosis of plasma cells. In this study we evaluated the characteristics of intracellular polyphosphate in myeloma cell lines, in primary myeloma cells from patients, and in other human B-cell populations from healthy donors. Design and MethodsWe have developed a novel flow cytometric method for detecting levels of polyphosphate in cell populations. We also used confocal microscopy and enzymatic analysis to study polyphosphate localization and characteristics. ResultsWe found that myeloma plasma cells contain higher levels of intracellular polyphosphate than normal plasma cells and other B-cell populations. Localization experiments indicated that high levels of polyphosphate accumulate in the nucleolus of myeloma cells. As the principal function of the nucleolus involves transcription of ribosomal DNA genes, we found changes in the cellular distribution of polyphosphate after the inhibition of nucleolar transcription. In addition, we found that RNA polymerase I activity, responsible for transcription in the nucleolus, is also modulated by polyphosphate, in a dose-dependent manner. ConclusionsOur results show an unusually high accumulation of polyphosphate in the nucleoli of myeloma cells and a functional relationship of this polymer with nucleolar transcription. Haematologica 2012;97(8):1264-1271. doi:10.3324/haematol.2011 This is an open-access paper. Myeloma cells contain high levels of inorganic polyphosphate which is associated with nucleolar transcription ABSTRACT© F e r r a t a S t o r t i F o u n d a t i o n

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Section: Polyphosphate Is Present In Myeloma Cell Linesmentioning

confidence: 98%

Section: Polyphosphate Is Present In Myeloma Cell Linesmentioning

confidence: 99%

Myeloma cells contain high levels of inorganic polyphosphate which is associated with nucleolar transcription

Jimenez-Nuñez¹,

Moreno-Sanchez²,

Hernández-Ruiz³

et al. 2012

Haematologica

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“…Fluorescence microscopy images showed an abundance of yellow fluorescent granules (Fig. 1A), corresponding to the shifting fluorescent emission of polyP after DAPI staining (10). To confirm the identity of the granules further, SEM-energy dispersive spectroscopy (EDS) was applied to the granules separated from sponge tissue by Percoll density gradient.…”

mentioning

confidence: 99%

Phosphorus sequestration in the form of polyphosphate by microbial symbionts in marine sponges

Zhang

Blasiak

Karolin

et al. 2015

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

127

112

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Marine sponges are major habitat-forming organisms in coastal benthic communities and have an ancient origin in evolution history. Here, we report significant accumulation of polyphosphate (polyP) granules in three common sponge species of the Caribbean coral reef. The identity of the polyP granules was confirmed by energy-dispersive spectroscopy (EDS) and by the fluorescence properties of the granules. Microscopy images revealed that a large proportion of microbial cells associated with sponge hosts contained intracellular polyP granules. Cyanobacterial symbionts cultured from sponges were shown to accumulate polyP. We also amplified polyphosphate kinase (ppk) genes from sponge DNA and confirmed that the gene was expressed. Based on these findings, we propose here a potentially important phosphorus (P) sequestration pathway through symbiotic microorganisms of marine sponges. Considering the widespread sponge population and abundant microbial cells associated with them, this pathway is likely to have a significant impact on the P cycle in benthic ecosystems.phosphorus sequestration | sponge | microbial symbionts | polyphosphate

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“…JC‐D8 and DAPI specifically label free polyP and have no selectivity towards purines, including ATP or ADP (Angelova et al, 2014; Aschar‐Sobbi et al, 2008). To study the functional roles of ATP or ADP as signaling molecules, enzymatic depletion of ATP or ADP by phosphatases has been used in a number of studies (Wells et al, 2015).…”

Section: Resultsmentioning

confidence: 99%

“…For images in experiments comparing levels of fluorescence in different cells, the imaging setting were kept at the same level. The DAPI‐polyP (which was separated from DAPI–DNA/RNA) fluorescence was detected with excitation 405 nm and emission between 480 and 520 nm according to (Aschar‐Sobbi et al, 2008). Mitochondrial localization was identified using potential sensitive indicator tetramethylrhodamine (TMRM; Angelova et al, 2018).…”

Section: Methodsmentioning

confidence: 99%

Signal transduction in astrocytes: Localization and release of inorganic polyphosphate

Angelova

Iversen

Teschemacher

et al. 2018

Glia

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Inorganic polyphosphate (polyP) is present in every cell and is highly conserved from primeval times. In the mammalian cells, polyP plays multiple roles including control of cell bioenergetics and signal transduction. In the brain, polyP mediates signaling between astrocytes via activation of purinergic receptors, however, the mechanisms of polyP release remain unknown. Here we report identification of polyP‐containing vesicles in cortical astrocytes and the main triggers that evoke vesicular polyP release. In cultured astrocytes, polyP was localized predominantly within the intracellular vesicular compartments which express vesicular nucleotide transporter VNUT (putative ATP‐containing vesicles), but not within the compartments expressing vesicular glutamate transporter 2 (VGLUT2). The number of lysosomes which contain polyP was dependent on the conditions of astrocytes. Release of polyP from a proportion of lysosomes could be induced by calcium ionophores. In contrast, polyP release from the VNUT‐containing vesicles could be triggered by various physiological stimuli, such as pH changes, polyP induced polyP release and other stimuli which increase [Ca2+]i. These data suggest that astrocytes release polyP predominantly via exocytosis from the VNUT‐containing vesicles. © 2018 Wiley Periodicals, Inc.

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High Sensitivity, Quantitative Measurements of Polyphosphate Using a New DAPI-Based Approach

Cited by 213 publications

References 29 publications

Myeloma cells contain high levels of inorganic polyphosphate which is associated with nucleolar transcription

Myeloma cells contain high levels of inorganic polyphosphate which is associated with nucleolar transcription

Phosphorus sequestration in the form of polyphosphate by microbial symbionts in marine sponges

Signal transduction in astrocytes: Localization and release of inorganic polyphosphate

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