Diversity of biological clocks in the tidal environment : molecular mechanism

河口蟹的卵附着系统是在产卵后母蟹多卵的毛发上形成的，孵化后会从这些毛发上滑落。这种剥离是由胚胎孵化后释放的一种活性因子OHSS(卵毛剥离物质)引起的，OHSS经过纯化后，其活性形式为25 kDa。从胚胎文库中克隆的OHSS全长为1759个氨基酸，在一个开放阅读框(ORF)中编码492个氨基酸。预测蛋白的C末端部分由类胰酶丝氨酸蛋白酶结构域组成，与其他动物的同源性为33%-38%。预测的蛋白质(54.7 kDa)以酶原的形式分泌，可能在翻译后被切割，将C-端与N-端区分开。两年前，我们发现了河口蟹孵化的奇妙现象。为了研究十足类甲壳类动物孵化的机制，我们尝试诱导…通过使用多种化学品进行更多的孵化。几乎所有的化学物质都是无效的，但他发现只有丙酮和乙腈溶液才能导致胚胎孵化。在对照水(4 0 0~5‰人工海水4 0 0~5 0 0ml5)曝气条件下孵化出的胚胎未脱离雌体。丙酮处理10min后约49h开始孵化，55h后孵化率达到最大。所有用丙酮处理过的动物园都在游泳，它们的行为与从雌性体内释放出来的动物园完全一样，孵化率因丙酮浓度的不同而有很大差异。用60%丙酮处理的胚胎孵化率为95%，用80%丙酮处理的胚胎孵化率为87%。因此，60%丙酮溶液的孵化率最高。在本研究的第一步，我们认为血吸虫胚胎孵化可能是由丙酮等物质引起的。孵化当然是由雌性释放的一种活跃因素引起的。但许多实验表明，孵化诱导物质与乙酰等化学物质有很大不同。我们正在分析这种物质。较少

The egg attachment system of an estuarine crab Sesarma haematocheir is formed on the maternal ovigerous hairs just after egg-laying, and slips off these hairs just after hatching. The stripping is caused by an active factor that we call OHSS (ovigerous-hair stripping substance), which is released by the embryo upon hatching.OHSS was purified, and its active form had a molecular mass of 25 kDa. The cDNA of OHSS cloned from an embryonic cDNA library was 1759 by long, encoding 492 amino acids in a single open reading frame (ORF). The C-terminal part of the predicted protein was composed of a trypsin-like serine protease domain, with homology to counterparts in other animals of 33-38%. The predicted protein (54.7 kDa) secreted as a zymogen may be cleaved post-translationally, separating the C-terminal from the N-terminal region.Two years before, we found fantastic phenomena about hatching of an estuarine crab. To investigate mechanisms of hatching in decapod crustaceans, we tried to induce … More hatching by use of many kinds of chemicals. Almost chemicals are ineffective, but he found that only acetone and acetonitrile solutions can cause hatching of embryos. No embryos detached from the female hatched under the control water with aeration (400-500 ml of 5‰ artificial seawater). Hatching started around 49 hrs after the treatment with acetone for 10 min, and hatching ratio attained maximum 55 hrs after. All zoeas treated with acetone swam, and their behavior was completely the same as zoeas released from the female.The ratio of hatching was largely different according to the concentration of acetone. Ninety-five percent hatched among the embryos treated with 60% acetone, and 87% hatched among those treated with 80% acetone. Thus, the solution of 60% acetone was estimated to cause maximum ratio of hatching.At the first step of this study, we thought that hatching of S. haematocheir embryos could be caused by a substance such as acetone. Hatching is certainly caused by an active factor released from the female. But many experiments suggested that the hatching inducing substance is largely different from a chemical such as acetolie. We are analyzing this substance. Less