Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
|Place of Conferral||海洋研究所|
|Keyword||长牡蛎 血糖 多元统计 转录组学|
为探究长牡蛎在自然生长周期内糖原含量和性腺发育等生理指标的变化，及其与类胰岛素基因（cgMIP123、cgMIP4、cgMILP7、cgILP）相对表达量的相关性，自2020年5月至2021年5月采集了山东胶南养殖海区的长牡蛎，测定了血糖、糖原含量、条件指数、目标基因相对表达量及环境因子（酸度、温度、盐度、叶绿素a含量）等相关数据，采用多元统计的方法对数据进行分析。logistic回归分析结果显示，长牡蛎配子排放前后血糖含量和内脏团糖原含量具有显著变化。构建的回归模型可以通过血糖含量和内脏团糖原含量准确判断配子是否已经排放，区分度C-index为0.903，Hosmer-Lemeshow拟合优度检验χ2值为9.06，p-value > 0.05，验证结果显示该模型可靠。多元线性回归分析结果显示：条件指数与cgMIP123、cgMIP4和cgMILP7具有相关性，R2为0.83，p-value为7.87e-04； cgMIP4和cgILP与糖原含量呈正相关；cgMIP123与糖原含量未显示出相关性；cgMILP7的相对表达量与内脏团和唇瓣组织的糖原含量及血糖含量呈负相关，推测该基因与性腺发育有关，在血糖调节中具有一定相关性但不是主要的调控因子。
The insulin-like superfamily is an ancient gene family in animals, which is involved in the regulation of a variety of physiological processes, including reproduction, development, growth, carbohydrate and lipid metabolism. There are many insulin-like peptide genes (ILPs) have been found in shellfish, but research on their function is scant and it is unclear what specific biological processes these genes are involved in. In this study, we selected Crassostrea gigas as experimental animal to assay the possible involvement of oyster ILPs in the regulation of glycogen content and gonad development. Physiological condition and gene expression levels were assayed on oysters collected monthly from the oyster farm, multivariate statistical analysis was then applied to study the correlations between ILP expression and physiological conditions. RNAseq was conducted on the hepatopancreas of oysters injected with glucose to assay the transcriptomic response to the blood glucose increase. The results are listed as follows:
Glycogen content variations during natural state of Crassostrea gigas and their correlations with the expression patterns of insulin-like genes (cgMIP123, cgMIP4, cgMILP7, cgILP). The blood glucose content, glycogen content, condition index and relative gene expression level were assayed on oysters cultured in a farm at Jiaonan, Shandong, while typical marine environmental factors (acidity, temperature, salinity, Chla) were collected from May 2020 to May 2021. Multivariate logistic regression analysis indicated high correlation between the spawning and variations of blood glucose content and glycogen content around pyloric process. A regression model was constructed to estimate the gonad discharging status by the two parameters. The concordance index was at 0.903, while the χ2 value was at 9.06 (p-value > 0.05) for Hosmer-Lemeshow test, suggesting high goodness of fit. Application of the model to verification samples also showed high reliability. The multiple linear regression analysis showed correlations of growth or physiological values with the expression pattern of some genes: the condition index was correlated with cgMIP123, cgMIP4 and cgMILP7 (R2: 0.83, p-value: 7.87e-04); cgMIP4 and cgILP were positively correlated with glycogen content; there is no correlation between cgMIP123 and glycogen content; the relative expression of cgMILP7 was negatively correlated with glycogen content of all the assayed tissues, this gene may be related to gonadal development and has partial correlation in blood glucose regulation.
After intramuscular injection of glucose, the blood glucose level of Crassostrea gigas increased rapidly in a short period of time, and then decreased gradually in a fluctuating way, and returned to the initial level 2 hours later. Transcriptome analysis of hepatopancreas of oysters after injection of glucose stimulation showed that insulin-like genes were not concentrated in differential genes, which may indicate that there was no change in insulin-like genes at the transcriptional level. cgPPP1R3B and cgPCSK1 were selected as two differential genes from the differential gene set. cgPPP1R3b played an important role in the process of glycogen synthesis, and cgPCSK1 plays an important role in the processing of ILP polypeptides, indicating enhanced regulation of ILP at the post-translational level. The results of starvation experiment did not show significant differences in blood glucose and glycogen content, which may be related to the short experiment period and fewer biological replicates.
|MOST Discipline Catalogue||理学::生物学|
|Table of Contents|
|苗校静. 长牡蛎类胰岛素基因与糖原含量和性腺发育的相关性研究[D]. 海洋研究所. 中国科学院大学,2021.|
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