Aquatic Molecular Genetics and Biotechnology Laboratory, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Klong Luang, Pathumthani, Thailand.
Mol Reprod Dev. 2011 May;78(5):347-60. doi: 10.1002/mrd.21310. Epub 2011 May 3.
Isolation and characterization of genes and/or proteins differentially expressed in ovaries are necessary for understanding ovarian development in the giant tiger shrimp (Penaeus monodon). In this study, the full-length cDNA of P. monodon mitogen-activating protein kinase 1 (PmMAPK1) was characterized. PmMAPK1 was 1,398 bp in length containing an open reading frame of 1,098 bp that corresponded to a polypeptide of 365 amino acids. PmMAPK1 was more abundantly expressed in ovaries than in testes of P. monodon. Quantitative real-time PCR revealed differential expression levels of PmMAPK1 mRNA during ovarian development of intact broodstock, where it peaked in early cortical rod (stage III) ovaries (P < 0.05) and slightly decreased afterwards (P > 0.05). Likewise, the expression level of PmMAPK1 in early cortical rod and mature (IV) ovaries was significantly greater than that in previtellogenic (I) and vitellogenic (II) ovaries of eyestalk-ablated broodstock (P < 0.05). The PmMAPK1 transcript was localized in ooplasm of previtellogenic oocytes. In intact broodstock, the expression of the PmMAPK1 protein was clearly increased from previtellogenic ovaries in subsequent stages of ovarian development (P < 0.05). In contrast, the level of ovarian PmMAPK1 protein was comparable during oogenesis in eyestalk-ablated broodstock (P > 0.05). The PmMAPK1 protein was localized in ooplasm of previtellogenic and vitellogenic oocytes. It was also detected around the nuclear membrane of early cortical rod oocytes in both intact and eyestalk-ablated broodstock. Results indicated that PmMAPK1 gene products seem to play functional roles in the development and maturation of oocytes/ovaries in P. monodon.
为了了解斑节对虾卵巢发育的机制,有必要分离和鉴定在卵巢中差异表达的基因和/或蛋白。本研究对斑节对虾丝裂原活化蛋白激酶 1(PmMAPK1)的全长 cDNA 进行了鉴定。PmMAPK1 全长 1398bp,开放阅读框为 1098bp,编码 365 个氨基酸的多肽。PmMAPK1 在斑节对虾卵巢中的表达量明显高于精巢。定量实时 PCR 结果显示,在完整亲虾卵巢发育过程中,PmMAPK1mRNA 的表达水平存在差异,在早期皮质棒状期(III 期)卵巢中表达水平最高(P<0.05),随后略有下降(P>0.05)。同样,早期皮质棒状期和成熟(IV 期)卵巢中 PmMAPK1 的表达水平显著高于经切除眼柄亲虾的前卵黄生成期(I 期)和卵黄生成期(II 期)卵巢(P<0.05)。PmMAPK1 转录本定位于前卵黄生成卵母细胞的卵质中。在完整亲虾中,PmMAPK1 蛋白的表达水平在卵巢发育的随后阶段明显增加(P<0.05)。相反,在切除眼柄亲虾的卵发生过程中,卵巢 PmMAPK1 蛋白水平相当(P>0.05)。PmMAPK1 蛋白定位于前卵黄生成期和卵黄生成期卵母细胞的卵质中,在完整和切除眼柄亲虾的早期皮质棒状期卵母细胞的核膜周围也检测到该蛋白。结果表明,PmMAPK1 基因产物可能在斑节对虾卵母细胞/卵巢的发育和成熟过程中发挥功能作用。