Jayasankar Vidya, Jasmani Safiah, Tsutsui Naoaki, Aida Katsumi, Wilder Marcy N
Japan International Research Center for Agricultural Sciences, 1-1 Ohwashi, Tsukuba, Ibaraki 305-8686, Japan.
J Exp Zool A Comp Exp Biol. 2006 May 1;305(5):440-8. doi: 10.1002/jez.a.272.
The dynamics of vitellogenin (Vg) mRNA expression and patterns of Vg and vitellin distribution in the hepatopancreas and ovary of juvenile Macrobrachium rosenbergii were examined using real-time RT-PCR and immunohistochemical methods. Eyestalk ablation was seen to induce rapid development of the gonads and Vg synthesis in females. In the female hepatopancreas, Vg mRNA expression was observed several days following ablation, after which levels increased gradually with increasing gonadosomatic index (GSI). Vitellin accumulation in the oocytes also increased with increasing Vg mRNA synthesis; expression was however negligible in the ovary. Hemolymph Vg levels in females ranged from 0.04 to 2.2 mg/ml. SDS PAGE/Western blotting analysis of hemolymph samples revealed that juvenile Vg was composed of 199 and 90 kDa subunits; the 102 kDa subunit present in adult female Vg (Okuno et al., 2002. J Exp Zool 292:417-429) could not be detected at any stage of vitellogenesis in juveniles. Vg was not detectable in non-ablated juveniles. The results of this study confirmed that the mode of involvement of eyestalk factors in regulating vitellogenesis is intrinsic to both juveniles and adults, and that a basic pattern of Vg synthesis and processing is conserved. However, the fact that juveniles are not able to produce the same Vg levels observed in adult females, and do not reach high GSI levels culminating in spawning suggests that other factors and physiological conditions specific to adult females are necessary to demonstrate full reproductive ability.
采用实时逆转录聚合酶链反应(RT-PCR)和免疫组织化学方法,研究了罗氏沼虾幼虾肝胰腺和卵巢中卵黄蛋白原(Vg)mRNA的表达动态以及Vg和卵黄磷蛋白的分布模式。结果发现,切除眼柄可诱导雌性罗氏沼虾性腺快速发育并合成Vg。在雌性罗氏沼虾的肝胰腺中,切除眼柄几天后可观察到Vg mRNA表达,之后随着性腺指数(GSI)的增加,其水平逐渐升高。卵母细胞中的卵黄磷蛋白积累也随着Vg mRNA合成的增加而增加;然而,卵巢中的表达可以忽略不计。雌性罗氏沼虾血淋巴中的Vg水平在0.04至2.2mg/ml之间。对血淋巴样本进行的十二烷基硫酸钠聚丙烯酰胺凝胶电泳/蛋白质免疫印迹分析显示,幼虾的Vg由199kDa和90kDa的亚基组成;在幼虾卵黄发生的任何阶段均未检测到成年雌性Vg中存在的102kDa亚基(Okuno等人,2002年。《实验动物学杂志》292:417-429)。在未切除眼柄的幼虾中未检测到Vg。本研究结果证实,眼柄因子参与调节卵黄发生的模式在幼虾和成年虾中都是固有的,并且Vg合成和加工的基本模式是保守的。然而,幼虾无法产生与成年雌性相同水平的Vg,并且不能达到导致产卵的高GSI水平,这表明成年雌性特有的其他因素和生理条件对于展现完全的繁殖能力是必要的。