Corley-Smith G E, Lim C J, Brandhorst B P
Institute of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, British Columbia, Canada.
Genetics. 1996 Apr;142(4):1265-76. doi: 10.1093/genetics/142.4.1265.
To help investigate the evolutionary origin of the imprinting (parent-of-origin mono-allelic expression) of paternal genes observed in mammals, we constructed haploid and diploid androgenetic zebrafish (Danio rerio). Haploid androgenotes were produced by fertilizing eggs that had been X-ray irradiated to eliminate the maternal genome. Subsequent inhibition of the first mitotic division of haploid androgenotes by heat shock produced diploid androgenotes. The lack of inheritance of maternal-specific DNA markers (RAPD and SSR) by putative diploid and haploid androgenotes confirmed the androgenetic origin of their genomes. Marker analysis was performed on 18 putative androgenotes (five diploids and 13 haploids) from six families. None of 157 maternal-specific RAPD markers analyzed, some of which were apparently homozygous, were passed on to any of these putative androgenotes. A mean of 7.7 maternal-specific markers were assessed per family. The survival of androgenetic zebrafish suggests that if paternal imprinting occurs in zebrafish, it does not result in essential genes being inactivated when their expression is required for development. Production of haploid androgenotes can be used to determine the meiotic recombination rate in male zebrafish. Androgenesis may also provide useful information about the mechanism of sex determination in zebrafish.
为了帮助研究在哺乳动物中观察到的父源基因印记(亲本来源的单等位基因表达)的进化起源,我们构建了单倍体和二倍体雄核发育斑马鱼(Danio rerio)。单倍体雄核发育个体是通过对经X射线照射以消除母本基因组的卵子进行受精而产生的。随后通过热休克抑制单倍体雄核发育个体的第一次有丝分裂,从而产生二倍体雄核发育个体。假定的二倍体和单倍体雄核发育个体缺乏母本特异性DNA标记(随机扩增多态性DNA和简单序列重复)的遗传,这证实了它们基因组的雄核发育起源。对来自六个家系的18个假定雄核发育个体(5个二倍体和13个单倍体)进行了标记分析。在分析的157个母本特异性随机扩增多态性DNA标记中,有些显然是纯合的,但没有一个传递给这些假定的雄核发育个体中的任何一个。每个家系平均评估7.7个母本特异性标记。雄核发育斑马鱼的存活表明,如果斑马鱼中发生父源印记,那么当发育需要某些基因表达时,印记不会导致这些必需基因失活。单倍体雄核发育个体的产生可用于确定雄性斑马鱼的减数分裂重组率。雄核发育还可能为斑马鱼性别决定机制提供有用信息。