Research Center for Marine Biology, Tohoku University, Asamushi, Aomori, Aomori 039-3501, Japan.
Dev Growth Differ. 2010 Feb;52(2):195-207. doi: 10.1111/j.1440-169X.2009.01154.x. Epub 2010 Jan 7.
We have cloned and studied Hp-ECPN, an encephalopsin orthologue of the sea urchin Hemicentrotus pulcherrimus. Hp-ecpn cDNA was produced and found to contain a 1461-bp open reading frame that encodes 486 amino acids. Accumulation of Hp-ecpn mRNA and protein expression occurred at the 14 h postfertilization (hpf) swimming blastula stage and thereafter. The Hp-ECPN protein was N-glycosylated, and the amino acid sequence was similar to that of vertebrate encephalopsins. Whole-mount immunohistochemistry revealed the presence of Hp-ECPN in cells (ECPN cells) that appeared initially around the tip of the archenteron in 20 hpf early gastrulae. By the 54 hpf pluteus stage, ECPN cells had spread through the aboral ectoderm, and, by the eight-arm pluteus stage, were restricted to the tips of the larval arms and the posterior end of the body. The number of ECPN cells increased under conditions of continuous light, but decreased under continuous dark. Knockdown of Hp-ecpn mRNA using morpholino antisense oligonucleotides decreased the number of ECPN cells considerably, and inhibited the vertical swimming of the larvae. This suggested that Hp-ECPN plays a role in photosensitive larval swimming vertical migration. In adult tissues, the ECPN cells were detected exclusively in tube feet.
我们已经克隆并研究了海胆(Hemicentrotus pulcherrimus)的视蛋白同源物 Hp-ECPN。Hp-ecpn cDNA 被产生并发现包含一个 1461 个碱基对的开放阅读框,编码 486 个氨基酸。Hp-ecpn mRNA 的积累和蛋白表达发生在受精后 14 小时(hpf)的游泳囊胚阶段,此后一直存在。Hp-ECPN 蛋白被 N-糖基化,其氨基酸序列与脊椎动物视蛋白相似。全胚胎免疫组织化学显示,在 20 hpf 早期原肠胚中,位于原肠顶端周围的细胞(ECPN 细胞)中存在 Hp-ECPN。到 54 hpf 变态期,ECPN 细胞已通过口后外胚层扩散,到 8 臂变态期,局限于幼虫臂的顶端和身体的后端。在连续光照条件下,ECPN 细胞的数量增加,但在连续黑暗条件下减少。使用反义寡核苷酸的 Hp-ecpn mRNA 敲低显著减少了 ECPN 细胞的数量,并抑制了幼虫的垂直游泳。这表明 Hp-ECPN 在感光幼虫垂直迁移的游泳中发挥作用。在成年组织中,ECPN 细胞仅在管足中被检测到。