Incorporation of nitrogen isotope 15N into liver DNA during avian embryogenesis: a new approach for measuring the rate of DNA synthesis

André Chwalibog; C.C. Metges; E. Sawosz Chwalibóg; M. Grodzik; T. Niemiec

Incorporation of nitrogen isotope ¹⁵N into liver DNA during avian embryogenesis: a new approach for measuring the rate of DNA synthesis

André Chwalibog, C.C. Metges, E. Sawosz Chwalibóg, M. Grodzik, T. Niemiec

Abstract

The objective of this experiment was to test the possibilities of measuring the rate of DNA synthesis in chicken embryos by applying a simple ¹⁵N tracer technique. We hypothesized that the rate of ¹⁵N incorporation into liver DNA depends on the type of labelled substance, reflecting precursor availability to provide substrates for nucleotide synthesis. Fertilized eggs were divided into 4 groups (4 × 15): control - not treated, and treated with ¹⁵N labeled glycine, ammonium chloride, or sodium nitrate. ¹⁵N labeled solutions were given in ovo by injection into albumen. After 20 days of incubation, the labeled substances had no effect on embryo development or morphology. Hepatic DNA was purified and ¹⁵N abundance was measured by isotope ratio mass spectrometry. There was significant enrichment of ¹⁵N in DNA from the glycine and ammonium chloride groups. We conclude that this simple technique of injecting ¹⁵N tracers into incubating eggs can be used to estimate the rate of DNA synthesis.

Original language	English
Journal	Journal of Animal and Feed Sciences
Volume	19
Pages (from-to)	269-276
Number of pages	8
ISSN	1230-1388
Publication status	Published - 2010

Cite this

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title = "Incorporation of nitrogen isotope 15N into liver DNA during avian embryogenesis: a new approach for measuring the rate of DNA synthesis",

abstract = "The objective of this experiment was to test the possibilities of measuring the rate of DNA synthesis in chicken embryos by applying a simple 15N tracer technique. We hypothesized that the rate of 15N incorporation into liver DNA depends on the type of labelled substance, reflecting precursor availability to provide substrates for nucleotide synthesis. Fertilized eggs were divided into 4 groups (4 × 15): control - not treated, and treated with 15N labeled glycine, ammonium chloride, or sodium nitrate. 15N labeled solutions were given in ovo by injection into albumen. After 20 days of incubation, the labeled substances had no effect on embryo development or morphology. Hepatic DNA was purified and 15N abundance was measured by isotope ratio mass spectrometry. There was significant enrichment of 15N in DNA from the glycine and ammonium chloride groups. We conclude that this simple technique of injecting 15N tracers into incubating eggs can be used to estimate the rate of DNA synthesis.",

author = "Andr{\'e} Chwalibog and C.C. Metges and Chwalib{\'o}g, {E. Sawosz} and M. Grodzik and T. Niemiec",

year = "2010",

language = "English",

volume = "19",

pages = "269--276",

journal = "Journal of Animal and Feed Sciences",

issn = "1230-1388",

publisher = "Polska Akademia Nauk Instytut Fizjologii i Zywienia Zwierzat im. Jana Kielanowskiego",

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TY - JOUR

T1 - Incorporation of nitrogen isotope 15N into liver DNA during avian embryogenesis

T2 - a new approach for measuring the rate of DNA synthesis

AU - Chwalibog, André

AU - Metges, C.C.

AU - Chwalibóg, E. Sawosz

AU - Grodzik, M.

AU - Niemiec, T.

PY - 2010

Y1 - 2010

N2 - The objective of this experiment was to test the possibilities of measuring the rate of DNA synthesis in chicken embryos by applying a simple 15N tracer technique. We hypothesized that the rate of 15N incorporation into liver DNA depends on the type of labelled substance, reflecting precursor availability to provide substrates for nucleotide synthesis. Fertilized eggs were divided into 4 groups (4 × 15): control - not treated, and treated with 15N labeled glycine, ammonium chloride, or sodium nitrate. 15N labeled solutions were given in ovo by injection into albumen. After 20 days of incubation, the labeled substances had no effect on embryo development or morphology. Hepatic DNA was purified and 15N abundance was measured by isotope ratio mass spectrometry. There was significant enrichment of 15N in DNA from the glycine and ammonium chloride groups. We conclude that this simple technique of injecting 15N tracers into incubating eggs can be used to estimate the rate of DNA synthesis.

AB - The objective of this experiment was to test the possibilities of measuring the rate of DNA synthesis in chicken embryos by applying a simple 15N tracer technique. We hypothesized that the rate of 15N incorporation into liver DNA depends on the type of labelled substance, reflecting precursor availability to provide substrates for nucleotide synthesis. Fertilized eggs were divided into 4 groups (4 × 15): control - not treated, and treated with 15N labeled glycine, ammonium chloride, or sodium nitrate. 15N labeled solutions were given in ovo by injection into albumen. After 20 days of incubation, the labeled substances had no effect on embryo development or morphology. Hepatic DNA was purified and 15N abundance was measured by isotope ratio mass spectrometry. There was significant enrichment of 15N in DNA from the glycine and ammonium chloride groups. We conclude that this simple technique of injecting 15N tracers into incubating eggs can be used to estimate the rate of DNA synthesis.

M3 - Journal article

SN - 1230-1388

VL - 19

SP - 269

EP - 276

JO - Journal of Animal and Feed Sciences

JF - Journal of Animal and Feed Sciences

ER -

Incorporation of nitrogen isotope 15N into liver DNA during avian embryogenesis: a new approach for measuring the rate of DNA synthesis

Abstract

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Incorporation of nitrogen isotope ¹⁵N into liver DNA during avian embryogenesis: a new approach for measuring the rate of DNA synthesis