Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, China.
Dev Comp Immunol. 2013 Sep;41(1):27-32. doi: 10.1016/j.dci.2013.04.005. Epub 2013 Apr 11.
An i-type lysozyme (MmeLys) has been proved to function in immunity of the clam Meretrix meretrix in our previous studies. In this study, the expression patterns of MmeLys mRNA and protein at four chief developmental stages of M. meretrix were analyzed, which was able to provide information about how ontogeny of immunity and, in particular, antibacterial ability occured in the bivalve. The results of real-time PCR and western blot showed that MmeLys expressions were activated in D-veligers and dramatically increased to the highest level in pediveligers. It is proposed that the expression changes at these two stages might be due to the visceral organs changes, which were related to the archenteron formation in D-veligers and the organ-restructuring in pediveligers during metamorphosis. In addition, new methods of whole mount in situ hybridization and whole mount immunofluorescence were applied to identify the MmeLys expression tissues, and these tissues (i.e. hepatopancreas, gill, mantle, mouth, velum and foot) may be involved in the immune function during development of clams. Our study is valuable to a certain extent for exploring the origin of immune functions in clams and provides new methodology for future studies on the immune ontogeny of bivalves.
在我们之前的研究中,已经证明 i 型溶菌酶(MmeLys)在贻贝 Meretrix meretrix 的免疫中发挥作用。在这项研究中,分析了 M. meretrix 四个主要发育阶段的 MmeLys mRNA 和蛋白质的表达模式,这为了解贝类的免疫发生和特别是抗菌能力的发生提供了信息。实时 PCR 和 Western blot 的结果表明,MmeLys 在 D 形幼虫中被激活,并在幼体中急剧增加到最高水平。据推测,这两个阶段的表达变化可能是由于内脏器官的变化所致,这些变化与 D 形幼虫中的原肠形成和变态期间幼体中的器官重构有关。此外,还应用了全胚胎原位杂交和全胚胎免疫荧光的新方法来鉴定 MmeLys 的表达组织,这些组织(即肝胰腺、鳃、套膜、嘴、翼和足)可能参与了贻贝发育过程中的免疫功能。我们的研究在一定程度上有助于探索贝类免疫功能的起源,并为未来研究贝类的免疫发生提供了新的方法。