Unit of Biochemistry, Department of Zoology, University of Madras, School of Life Science, Guindy, Chennai, Tamil Nadu, India.
J Exp Zool B Mol Dev Evol. 2012 Nov;318(7):572-85. doi: 10.1002/jez.b.22462. Epub 2012 Jul 13.
Vitellogenin (VTG) synthesis in the hepatopancreas and ovary is negatively regulated by vitellogenesis-inhibiting hormone (VIH) produced in the neurosecretory cell of X-organ/sinus gland complex of the eyestalks of penaeid shrimp. Eyestalk ablation is used commercially to induce ovarian maturation in shrimps which leads to an eventual loss of the spawner. The aim of the present study was to understand the molecular mechanism of VIH regulation in ovarian development and its inhibition of VTG gene expression by using a MEK-specific inhibitor (U0126). The real-time quantitative PCR results showed VTG mRNA level was progressively increased in the ovary and hepatopancreas of unilateral eyestalk-ablated and inhibitor-treated shrimps. Western blot analysis also showed that phosphoMEK was detected only in the unilateral eyestalk-ablated and control shrimp, whereas phospho-MEK was not detected in inhibitor-treated shrimp. DAX-1, SF-1, and StAR expression correlated with changes in VIH mRNA and altered phospho-ERK levels. This is consistent with the hypothesis that suppression of DAX-1 results in SF-1-mediated StAR protein upregulation of estradiol that is implicated in vitellogenesis. This is the first report that demonstrates the molecular mechanism of VIH suppression via MEK pathway to induce ovarian maturation in female Penaeus monodon by molecular signal intervention, a less-invasive method than traditional eyestalk ablation.
在对虾的眼柄神经分泌细胞中产生的卵黄生成抑制激素(VIH)负调控肝胰腺和卵巢中的卵黄原蛋白(VTG)合成。眼柄切除被商业上用于诱导虾的卵巢成熟,这最终导致产卵者的损失。本研究的目的是通过使用 MEK 特异性抑制剂(U0126)来了解 VIH 在卵巢发育中的调节分子机制及其对 VTG 基因表达的抑制作用。实时定量 PCR 结果显示,单侧眼柄切除和抑制剂处理的虾的卵巢和肝胰腺中的 VTG mRNA 水平逐渐增加。Western blot 分析还表明,仅在单侧眼柄切除和对照虾中检测到磷酸化 MEK,而在抑制剂处理的虾中未检测到磷酸化 MEK。DAX-1、SF-1 和 StAR 的表达与 VIH mRNA 的变化和改变的磷酸化 ERK 水平相关。这与抑制 DAX-1 导致 SF-1 介导的 StAR 蛋白上调雌二醇从而参与卵黄发生的假说一致。这是第一个通过分子信号干预证明 MEK 途径抑制 VIH 以诱导雌性斑节对虾卵巢成熟的分子机制的报告,这是一种比传统眼柄切除更微创的方法。