Johnston Stephen D, López-Fernández Carmen, Arroyo Francisca, Gosálbez Altea, Cortés Gutiérrez Elva I, Fernández Jose-Luis, Gosálvez Jaime
School of Agriculture and Food Science, The University of Queensland, Gatton, Queensland, 4343, Australia.
Faculty of Biology, Autonomous University of Madrid, Cantoblanco, Madrid, Spain.
J Assist Reprod Genet. 2016 Sep;33(9):1231-8. doi: 10.1007/s10815-016-0754-9. Epub 2016 Jun 20.
Using a rabbit model, we assessed the influence of sperm DNA longevity on female reproductive outcomes.
Semen was collected from 40 bucks, incubated at 38 °C for 24 h, and the rate of sperm DNA fragmentation (rSDF) was determined using the sperm chromatin dispersion assay. Males were allocated into high rSDF (>0.5 units of increase per hour) or low rSDF (<0.5 units of increase per hour) groups. High and low rSDF semen samples were sequentially artificially inseminated into the same doe to reduce female factor variability, and pregnancy outcomes were recorded.
While there was no difference in SDFs between rSDF groups immediately after collection (T0), differences were significant after 2 h of incubation; SDFs determined at collection and rSDF behaved as independent characters (Pearson correlation = 0.099; P = 0.542). Following artificial insemination, the rate of stillborn pups was significantly higher in does inseminated by males with a high rSDF (14/21) compared to those with low rSDF (15/6); (contingency χ(2) 5.19; p = 0.022). The risk of stillborn when low rSDF rabbits were used for insemination was 0.16, but increased to 0.36 when high rSDF animals were used (odds ratio = 2.85; 95 % confidence interval = 1.4-2.7).
CONCLUSION(S): Dynamic assessment of SDF coupled with natural multiple ovulation, high fecundity of the rabbit and control over female factor influence, provided a useful experimental model to demonstrate the adverse effect of reduced sperm DNA longevity on reproductive outcome.
我们使用兔模型评估精子DNA寿命对雌性生殖结局的影响。
从40只雄兔采集精液,在38°C孵育24小时,采用精子染色质扩散试验测定精子DNA碎片率(rSDF)。将雄兔分为高rSDF组(每小时增加>0.5单位)或低rSDF组(每小时增加<0.5单位)。将高rSDF和低rSDF精液样本依次人工授精到同一只母兔体内,以减少雌性因素的变异性,并记录妊娠结局。
采集后即刻(T0),rSDF组之间的精子DNA碎片率无差异,但孵育2小时后差异显著;采集时测定的精子DNA碎片率和rSDF表现为独立性状(Pearson相关性=0.099;P=0.542)。人工授精后,高rSDF雄兔授精的母兔死产率(14/21)显著高于低rSDF雄兔授精的母兔(15/6);(列联χ(2) 5.19;p=0.022)。低rSDF兔用于授精时死产风险为0.16,但高rSDF兔用于授精时风险增加至0.36(优势比=2.85;95%置信区间=1.4-2.7)。
对精子DNA碎片率进行动态评估,结合兔的自然多次排卵、高繁殖力以及对雌性因素影响的控制,提供了一个有用的实验模型,以证明精子DNA寿命缩短对生殖结局的不利影响。