菌胁迫对近江牡蛎C型凝集素母源免疫传递及抑菌影响

Maternal Immune Transmission and Bacteriostatic Effect of Bacterial Stress on C-type Lectin in Crassostrea rivularis

为丰富无脊椎贝类母源免疫调控机制研究,文章采用底物显色原位杂交、荧光定量PCR和人工促排等技术,研究了菌胁迫对近江牡蛎(Crassostrearivularis)C型凝集素(C-typelectin,CTL)母源免疫传递及抑菌影响。结果表明:CTL定位于成贝卵巢组织的滤泡细胞和卵原细胞中;成贝卵巢组织和成熟卵子中均能检测到CTL转录本;成贝注射金黄色葡萄球菌或大肠杆菌后,卵巢组织CTLmRNA表达量均有一个持续上升过程,注射后12h的CTLmRNA表达量达最高峰,注射后48h大致恢复至空白组水平;成贝注射金黄色葡萄球菌或大肠杆菌12h后,人工促排,与空白组比较,实验组成熟卵子中CTLmRNA相对表达量均显著上调(P<0.05);抑菌活性实验显示成熟卵子蛋白具有抑菌活性,且抑菌强度随孵育时间的延长和蛋白浓度的增加而增强,但其中的CTL与其抗体结合后,抑菌活性下降,菌存活率显著上调(P<0.05)。该CTL在近江牡蛎中具有明显的母源免疫传递效应,且成熟卵子中母源CTL具有重要的抑菌活性。

In order to enrich the research on the maternal immune regulation mechanism of invertebrate shellfish,influence of bacterial stress on C-type lectin maternal immunity transmission and bacteriostasis in Crassostrea rivularis,this paper studied the using technology of substrate chromogenic in situ hybridization,real-time PCR,artificial ovulation promotion.Results showed that CTL was located in follicular cell and oogonia of adult shellfish ovary tissue,ovarian tissue and mature eggs had expressions of CTL transcript,after injection of staphylococcus aureus or escherichia coli,relative expressions of CTL mRNA had a continuous rising process in the ovary tissue of adult shellfish,relative expressions of CTL mRNA reached the peak after injection 12h,and returned to the level of the blank group after 48h;after injection 12h of staphylococcus aureus or escherichia coli,artificial ovulation promotion was implemented,compared with the blank group,relatives expressions of CTL mRNA were significantly increased in mature eggs of the experimental group(P<0.05),the bacteriostatic activity experiment showed that mature egg protein had bacteriostatic activity,and the inhibition increased with the prolongation of incubation time and the increase of protein concentration,however,after CTL was combined with its antibody,the bacteriostatic activity decreased,and the bacterial survival rates were significantly increased(P<0.05).The CTL has obvious maternal immune transmission effect,and the maternal CTL has important antibacterial activity in mature eggs in Crassostrea rivularis.