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Comparing DNA enrichment of proliferating cells following administration of different stable isotopes of heavy water.

ABSTRACT: Deuterated water (²H₂O) is a label commonly used for safe quantitative measurement of deuterium enrichment into DNA of proliferating cells. More recently, it has been used for labeling proteins and other biomolecules. Our in vitro - in vivo research reports important stable isotopic labeling enrichment differences into the DNA nucleosides and their isotopologues (e.g. deoxyadenosine (dA) M?+?1, dA M?+?2, dA M?+?3), as well as tumor cell proliferation effects for various forms of commercially available stable heavy water (²H₂O, H₂¹⁸O, and ²H₂¹⁸O). Using an in vitro mouse thymus tumor cell line, we determined that H₂¹⁸O provides superior DNA labeling enrichment quantitation, as measured by GC-positive chemical ionization (PCI)-MS/MS. In addition, at higher but physiologically relevant doses, both ²H₂¹⁸O and ²H₂O down modulated mouse thymus tumor cell proliferation, whereas H₂¹⁸O water had no observable effects on cell proliferation. The in vivo labeling studies, where normal mouse bone marrow cells (i.e. high turnover) were evaluated post labeling, demonstrated DNA enrichments concordant with measurements from the in vitro studies. Our research also reports a headspace-GC-NCI-MS method, which rapidly and quantitatively measures stable heavy water levels in total body water.

SUBMITTER: Farthing DE

PROVIDER: S-EPMC5481421 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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Comparing DNA enrichment of proliferating cells following administration of different stable isotopes of heavy water.

Farthing Don E DE Buxbaum Nataliya P NP Lucas Philip J PJ Maglakelidze Natella N Oliver Brittany B Wang Jiun J Hu Kevin K Castro Ehydel E Bare Catherine V CV Gress Ronald E RE

Scientific reports 20170622 1

Deuterated water (<sup>2</sup>H<sub>2</sub>O) is a label commonly used for safe quantitative measurement of deuterium enrichment into DNA of proliferating cells. More recently, it has been used for labeling proteins and other biomolecules. Our in vitro - in vivo research reports important stable isotopic labeling enrichment differences into the DNA nucleosides and their isotopologues (e.g. deoxyadenosine (dA) M + 1, dA M + 2, dA M + 3), as well as tumor cell proliferation effects for various for ...[more]

PMID: 28642474

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Project description:Ecohydrological isotope based field research is often constrained by a lack of temporally explicit soil water data, usually related to the choice of destructive sampling in the field and subsequent analysis in the laboratory. New techniques based on gas permeable membranes allow to sample soil water vapor in situ and infer soil liquid water isotopic signatures. Here, a membrane-based in situ soil water vapor sampling method was tested at a grassland site in Freiburg, Germany. It was further compared with two commonly used destructive sampling approaches for determination of soil liquid water isotopic signatures: cryogenic vacuum extraction and centrifugation. All methods were tested under semi-controlled field conditions, conducting an experiment with dry-wet cycling and two isotopically different labeling irrigation waters. We found mean absolute differences between cryogenic vacuum extraction and in situ vapor measurements of 0.3-14.2‰ (δ18O) and 0.4-152.2‰ (δ2H) for soil liquid water. The smallest differences were found under natural abundance conditions of 2H and 18O, the strongest differences were observed after irrigation with labeled waters. Labeling strongly increased the isotopic variation in soil water: Mean soil water isotopic signatures derived by cryogenic vacuum extraction were -11.6 ± 10.9‰ (δ18O) and +61.9 ± 266.3‰ (δ2H). The in situ soil water vapor method showed isotopic signatures of -12.5 ± 9.4‰ (δ18O) and +169.3 ± 261.5‰ (δ2H). Centrifugation was unsuccessful for soil samples due to low water recovery rates. It is therefore not recommended. Our study highlights that the in situ soil water vapor method captures the temporal dynamics in the isotopic signature of soil water well while the destructive approach also includes the natural lateral isotopic heterogeneity. The different advantages and limitations of the three methods regarding setup, handling and costs are discussed. The choice of method should not only consider prevailing environmental conditions but the experimental design and goal. We see a very promising tool in the in situ soil water vapor method, capturing both temporal developments and spatial variability of soil water processes.

Dataset Information

Comparing DNA enrichment of proliferating cells following administration of different stable isotopes of heavy water.

Publications

Comparing DNA enrichment of proliferating cells following administration of different stable isotopes of heavy water.

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