Girish B P, Swetha C H, Reddy P Sreenivasula
Naturwissenschaften. 2015 Apr;102(3-4):20. doi: 10.1007/s00114-015-1272-9.
The objective of the present study was to investigate the expression profile of retinoid X receptor (RXR), ecdysone receptor (EcR) and ecdysone inducible gene (E75) in the hepatopancreas and ovary of Oziothelphusa senex senex during different vitellogenic stages. RXR, EcR and E75 complementary DNAs (cDNAs) were isolated from the ovaries, while vitellogenin (VtG) cDNA was isolated from the hepatopancreas of vitellogenic female crab. Deduced amino acid sequence of the messenger RNAs (mRNAs) of RXR, EcR and E75 showed more than 80% identity with their respective mRNAs of other brachyurans. VtG mRNA was not detected in the ovary throughout vitellogenic stages. RXR and EcR were significantly increased in the ovaries during vitellogenic stage I. The levels of EcR, E75 and VtG in the hepatopancreas elevated significantly during vitellogenic stages I and II, whereas the levels of RXR elevated only in vitellogenic stage I. During vitellogenic stage III, the levels of RXR, EcR and VtG in the hepatopancreas were significantly decreased. Immunoprecipitation analysis revealed the presence of VtG in the haemolymph, hepatopancreas and ovary extracts from the females but absent in haemolymph and hepatopancreas extract of males. It can be inferred that RXR, EcR and E75 are involved in the regulation of synthesis of VtG in hepatopancreas, whereas in ovary, it is hypothesized that they play an important role in the uptake of VtG from the haemolymph, probably by regulating the levels of vitellogenin receptor. These are the first data showing an association between the expression levels of RXR, EcR and E75 and vitellogenesis and provide an alternative molecular intervention mechanism to the traditional eyestalk ablation to induce vitellogenesis and ovarian maturation in crustaceans.
本研究的目的是调查黄斑沼虾在不同卵黄发生阶段肝胰腺和卵巢中视黄酸X受体(RXR)、蜕皮激素受体(EcR)和蜕皮激素诱导基因(E75)的表达谱。从卵巢中分离出RXR、EcR和E75互补DNA(cDNA),而从处于卵黄发生期的雌蟹肝胰腺中分离出卵黄蛋白原(VtG)cDNA。RXR、EcR和E75信使RNA(mRNA)推导的氨基酸序列与其各自在其他短尾类动物中的mRNA显示出超过80%的同一性。在整个卵黄发生阶段,卵巢中均未检测到VtG mRNA。在卵黄发生I期,卵巢中的RXR和EcR显著增加。在卵黄发生I期和II期,肝胰腺中EcR、E75和VtG的水平显著升高,而RXR的水平仅在卵黄发生I期升高。在卵黄发生III期,肝胰腺中RXR、EcR和VtG的水平显著下降。免疫沉淀分析显示,雌性的血淋巴、肝胰腺和卵巢提取物中存在VtG,而雄性的血淋巴和肝胰腺提取物中不存在。可以推断,RXR、EcR和E75参与肝胰腺中VtG合成的调节,而在卵巢中,推测它们可能通过调节卵黄蛋白原受体的水平在从血淋巴中摄取VtG方面发挥重要作用。这些是首批显示RXR、EcR和E75表达水平与卵黄发生之间存在关联的数据,并为传统的眼柄切除诱导甲壳类动物卵黄发生和卵巢成熟提供了一种替代的分子干预机制。