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

• Autorzy: Chwalibog A. , Metges C.C. , Sawosz Chwalibog E. , Grodzik M. , Niemiec T.
• Języki publikacji: EN

Abstrakty

EN

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.

Słowa kluczowe

PL

nitrogen isotope; stable isotope; DNA synthesis; liver; chicken; embryo; avian embryogenesis; embryogenesis; embryo development; morphology

• Wydawca: -
• Czasopismo: Journal of Animal and Feed Sciences
• Rocznik: 2010
• Tom: 19
• Numer: 2
• Opis fizyczny: p.269-276,fig.,ref.

Twórcy

autor

Chwalibog A.

• University of Copenhagen, Gronnegardsvej 3, 1870 Frederiksberg C, Denmark

autor

Metges C.C.

autor

Sawosz Chwalibog E.

autor

Grodzik M.

autor

Niemiec T.

Bibliografia

• Acquisti C., Elser J.J., Kumar S., 2009. Ecological nitrogen limitation shapes the DNA composition of plant genomes. Mol. Biol. Evol. 26, 953-956
• Backes G., Hennig U., Petzke K.J., Elsner A., Junghans P., Nürnberg G., Metges C.C., 2002. Contribution of intestinal microbial lysine to lysine homeostasis is reduced in minipigs fed a wheat gluten based diet. Amer. J. Clin. Nutr. 76, 1317-1325
• Boza J.J., Jahoor F., Reeds P.J., 1996. Ribonucleic acid nucleotides in maternal and fetal tissues derive almost exclusively from synthesis de novo in pregnant mice. J Nutr. 126, 1749-1758
• Chen P., Abramson F.P., 1998. Measuring DNA sunthesis rates with [1-13C] glycine. Anal. Chem. 70, 1664-1669
• Chivian D., Brodie E.L., Alm E.J. et al., 2008. Environmental genomics reveals a single-species ecosystem deep within earth. Science 322, 275-278
• Davidson J.N., 1946. Modern methods of studying protein metabolism. Proc. Nutr. Soc. 4, 249-253
• Fan M.Z., Chiba L.I., Matzat P.D., Yang X., Yin Y.L., Mine Y., Stein H.H., 2006. Measuring synthesis rates of nitrogen-containing polymers by using stable isotope tracers. J. Anim. Sci. 84, E79-E83
• Hamburger V., Hamilton H.L., 1951. A series of normal stages in the development of the chick embryo. J. Morphol. 88, 49-92
• Hellerstein M.K., 2003. In vivo measurement of fluxes through metabolic pathways: The missing link in functional genomics and pharmaceutical research. Annu. Rev. Nutr. 23, 379-402
• Martini W.Z., Chinkes D.L., Wolfe R.R., 2004. Quantification of DNA synthesis from different pathways in cultured human fibroblasts and myocytes. Metabolism 53, 128-133
• Metges C.C., Petzke K.J., 1997. Measurement of 15N/14N isotopic composition in individual plasma free amino acids of human adults at natural abundance by gas chromatography-combustion isotope ratio mass spectrometry. Anal. Biochem. 247, 158-164
• Metges C.C., Petzke K.J., El-Khoury A.E., Henneman L., Grant I., Bedri S., Regan M., Fuller M.F., Young V.R., 1999. Incorporation of urea and ammonia N into ileal and fecal microbial protein amino acids, and free plasma amino acids in normal men and ileostomates. Amer. J. Clin. Nutr. 70, 1046-1058
• Neese R.A., Siler S.Q., Cesar D., Antelo F., Lee D., Misell L., Patel K., Tehrani S., Shah P., Hellerstein M.K., 2001. Advances in the stable isotope-mass spectrometric measurement of DNA synthesis and cell proliferation. Anal. Biochem. 298, 189-195
• Nissim I., Starr S.E., Sullivan K.E., Campbell D.E., Douglas S.D., Daikhin Y., Nissim Il., Yudkoff M., 2000. Rapid method for determining the rate of DNA synthesis and cellular proliferation. Anal. Biochem. 278, 198-205
• Petzke K.J., Boeing H., Klaus S., Metges C.C., 2005. Carbon and nitrogen stable isotopic composition of hair protein and amino acids can be used as biomarkers for animal-derived dietary protein intake in humans. J. Nutr. 135, 1515-1520
• Perez J.F., Reeds P.J., 1998. A new stable isotope method enables the simultaneous measurement of nucleic acid and protein synthesis in vivo in mice. J. Nutr. 128, 1562-1569
• SAS, 1990. SAS Procedures Guide. Version 6, SAS Institute Inc. 3rd Edition. Cary NC
• Sawosz E., Grodzik M., Zielinska M., Niemiec T., Olszanska B., Chwalibog A., 2009. Nanoparticles of silver do not affect growth, development and DNA oxidative damage in chicken embryos. Arch. Geflügelk. 73, 208-213
• Zhang X.-J., Chinkes D.L., Wu Z., Martini W.Z., Wolfe R.R., 2004. Fractional synthesis rates of DNA and protein in rabbit skin are not correlated. J. Nutr. 134, 2401-2406