Araki K, Okamoto H, Graveson A C, Nakayama I, Nagoya H
Cell Engineering Section, Inland Station, National Research Institute of Aquaculture, Tamaki, Watarai, Mie 519-0423, Japan.
Dev Growth Differ. 2001 Oct;43(5):591-9. doi: 10.1046/j.1440-169x.2001.00601.x.
The abnormalities of haploid medaka embryos were characterized by comparative analysis of histologic sections and expression patterns of some developmental marker genes between haploids and diploids to clarify whether medaka haploids are useful for identifying mutants. During gastrulation, an obvious defect was first observed as a delay of epiboly and involution. This delay was shown to be caused not by the perturbation of mesoderm induction, but by widespread cell death and disorganization of cell arrangement in the blastoderm. This disorganization of cell arrangement was also detected in various organs, such as the brain, somite and notochord, at a late developmental stage. Ten days after fertilization, a small head and a short body axis were formed; these changes were also observed in haploid embryos in other species, but their cause is unknown. Based on the expression patterns of HNF3beta and goosecoid, it was demonstrated that a short and impotent prechordal plate induced near the marginal zone in haploid embryos was responsible for this defect. However, in these experiments it was also demonstrated that many major organs in haploids, such as the somite and notochord, differentiated incompletely but were present. Therefore, it was concluded that haploid screening is suitable for identifying mutations revealed by an obvious phenotype, such as dorsoventral polarity.
通过对单倍体和二倍体青鳉胚胎组织切片及一些发育标记基因表达模式的比较分析,来表征单倍体青鳉胚胎的异常情况,以阐明青鳉单倍体是否有助于鉴定突变体。在原肠胚形成过程中,首先观察到一个明显的缺陷,即外包和内卷延迟。结果表明,这种延迟不是由中胚层诱导的扰动引起的,而是由胚盘广泛的细胞死亡和细胞排列紊乱导致的。在发育后期,在脑、体节和脊索等各种器官中也检测到了这种细胞排列紊乱。受精十天后,形成了小头和短体轴;在其他物种的单倍体胚胎中也观察到了这些变化,但其原因尚不清楚。基于HNF3β和 goosecoid的表达模式,证明单倍体胚胎边缘区附近诱导形成的短而无力的脊索前板是造成这种缺陷的原因。然而,在这些实验中还证明,单倍体中的许多主要器官,如体节和脊索,虽然分化不完全,但仍然存在。因此,得出结论,单倍体筛选适用于鉴定由明显表型揭示的突变,如背腹极性。