Pilon M, Weisblat D A
Department of Molecular and Cell Biology, University of California, Berkeley 94720-3200, USA.
Development. 1997 May;124(9):1771-80. doi: 10.1242/dev.124.9.1771.
From the glossiphoniid leech Helobdella robusta, we have cloned and determined the complete coding sequence of Hro-nos, a gene homologous to the nanos gene from Drosophila melanogaster. Developmental northern blots show that Hro-nos, like nanos, is a maternal transcript that decays rapidly during early development. A polyclonal antiserum raised against the HRO-NOS protein was used in developmental western blots and for immunostaining leech embryos of different developmental stages. The HRO-NOS protein is first detectable in 2-cell embryos (4-6 hours of development) and exhibits a transient expression peaking during fourth cleavage (9-12 cells; 8-14 hours of development). The HRO-NOS protein exhibits a graded distribution along the primary embryonic axis and is partitioned unequally between the sister cells DNOPQ and DM, progeny of macromere D' at fourth cleavage: DNOPQ is the segmental ectoderm precursor cell and exhibits levels of HRO-NOS protein that are at least two-fold higher than in cell DM, the segmental mesoderm precursor cell. The observed expression pattern suggests that Hro-nos plays a role in the decision between ectodermal and mesodermal cell fates in leech.
我们从粗壮舌蛭(Helobdella robusta)中克隆并测定了Hro-nos的完整编码序列,该基因与黑腹果蝇(Drosophila melanogaster)的nanos基因同源。发育Northern印迹显示,Hro-nos与nanos一样,是一种母体转录本,在早期发育过程中迅速降解。用针对HRO-NOS蛋白制备的多克隆抗血清进行发育Western印迹分析,并用于对不同发育阶段的水蛭胚胎进行免疫染色。HRO-NOS蛋白最早在2细胞胚胎(发育4 - 6小时)中可检测到,并在第四次卵裂期间(9 - 12个细胞;发育8 - 14小时)呈现短暂表达峰值。HRO-NOS蛋白沿胚胎主轴呈梯度分布,并且在第四次卵裂时,在大卵裂球D'的子细胞DNOPQ和DM之间分配不均:DNOPQ是体节外胚层前体细胞,其HRO-NOS蛋白水平至少比体节中胚层前体细胞DM高两倍。观察到的表达模式表明,Hro-nos在水蛭外胚层和中胚层细胞命运的决定中起作用。