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NAD turnover during early development of Xenopus laevis
Authors:Gwyn T Williams  Sydney Shall  Christopher C Ford
Institution:Cell and Molecular Biology Laboratory, University of Sussex, Brighton, Sussex BN1 9QG U.K.
Abstract:The NAD pools of Xenopus laevis oocytes and early embryos can be radioactively labelled by microinjection of adenine- 3H]NAD. This technique is used to study the metabolism of NAD in oocytes and during early development. The rate at which NAD is degraded in vivo has been monitored by determining the rate of transfer of adenine residues from the NAD pool into other nucleotides and polynucleotides. In oocytes, NAD turnover is extremely slow, with a half-life of about 400 h. NAD turnover increases dramatically after fertilisation, and the half-life of the compound decreases to 37 h in 5-h-old embryos and to 10 h in 40-h-old embryos. 2 mM 3-aminobenzamide, a specific inhibitor of poly(ADP-ribose) polymerase, reduces the NAD turnover rate by about 20%, whereas 5 mM isonicotinic acid hydrazide, a specific inhibitor of NAD glycohydrolase, produces no significant inhibition. This indicates that a significant fraction of the considerable NAD turnover observed involves poly(ADP-ribose) polymerase. Our results indicate that poly(ADP-ribose) polymerase is active during early development and suggest that this activity may be involved in one or more aspects of the nuclear metabolism of the embryo.
Keywords:NAD turnover  Development  (X  laevis embryo)  Hepes  4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid  PPO  2′5-diphenyloxazole
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