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First published online February 27, 2009
Journal of Experimental Biology 212, 785-789 (2009)
Published by The Company of Biologists 2009
doi: 10.1242/jeb.023663
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Ametabolic embryos of Artemia franciscana accumulate DNA damage during prolonged anoxia

Alexander G. McLennan

Cell Regulation and Signalling Division, School of Biological Sciences, University of Liverpool, Liverpool L69 7ZB, UK


Figure 1
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  		Fig. 1. Time-dependent accumulation of abasic (AP) sites in the DNA of hydrated Artemia cysts (closed circles) and purified calf thymus DNA (open circles) stored under anoxic conditions at 28°C. For each determination, DNA was prepared from two separate 0.5 g cyst or 250 µl calf thymus DNA portions and duplicate ARP-DNA samples made from each of these and assayed in triplicate (N=4). Error bars are ± s.d. Comparison of the slopes of the two experimental regression lines using GraphPad Prism software showed the difference to be extremely significant (P<0.0001). The dotted line represents the predicted rate of depurination of DNA at pH 6.3 calculated from the data of Lindahl and Nyberg (Lindahl and Nyberg, 1972Go).

 

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  		Fig. 2. Time-dependent accumulation of abasic (AP) sites in the DNA of hydrated Artemia cysts (closed circles) and purified calf thymus DNA (open circles) stored under anoxic conditions at 40°C. Other details and statistical analysis as for Fig. 1.

 

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  		Fig. 3. Loss of AP sites from DNA of cysts and larvae hatched from cysts stored under anoxic conditions at 28°C for 30 weeks followed by incubation in oxygenated seawater at the same temperature. Black bars indicate the number of AP sites in cysts after 0, 12 and 24 h incubation in oxygenated seawater and in separated larvae hatched 48 and 72 h after introduction to oxygenated seawater. Error bars are ± s.d. (N=4, as in Fig. 1). Asterisks denote t-test significance compared with 0 h cysts (*P=0.01; **P=0.005; ***P=0.001). Grey bars indicate the % hatch of these cysts, determined on samples of 250–300 cysts distributed among the wells of a 24-well plate as previously described (Clegg, 2007Go).

 

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© The Company of Biologists Ltd 2009