| Ovarian development is a complex physiological process for crustacean reproduction that is divided into the oogonium proliferation stage,endogenous vitellogenic stage,exogenous vitellogenic stage,and oocyte maturation stage.Crustacean ovarian development involves the physiological processes of oogonia proliferation,oocyte differentiation,vitellogenesis and nutrient accumulation in oocytes,which are usually regulated by reproductive endocrine hormones,nutritional regulation and environmental factors.Chinese mitten crab(Eriocheir sinensis)is one of the important economic crabs in China.The status of ovarian development directly affects the quality and subsequent reproduction of adult E.sinensis.Therefore,it is very important to understand the ovarian development mechanism of E.sinensis.In view of this,this study used omics technology to study the changes of key genes,proteins and lipids molecules during ovarian development in E.sinensis,and discussed the regulatory mechanism of ovarian development from different levels,in order to provide reference for in-depth understanding of the molecular regulatory pathways and physiological processes of crustacean ovarian development.The main findings were as follows:1.Comparative transcriptome analysis reveals the dynamics of ovarian development in E.sinensisThis study performed transcriptome analysis in the ovary and hepatopancreas of E.sinensis during ovarian stages I(oogonium proliferation),II(endogenous vitellogenesis)and III(exogenous vitellogenesis).The results showed that a total of 5520 and 226 genes were expressed differently in ovary and hepatopancreas,respectively.For KEGG enrichment analysis,the differentially expressed genes(DEGs)in the ovary were significantly clustered in phototransduction-fly,phagosome and ECM-receptor interaction.The significantly enriched pathways in the hepatopancreas included fatty acid biosynthesis,fatty acid metabolism and riboflavin metabolism.Further analysis showed that twenty-five genes and several pathways were mainly involved in the process of oogenesis,including the ubiquitin/proteasome pathway(UPP),cyclic AMP-protein kinase A(c AMP-PKA)signaling pathway and mitogen-activated protein kinase(MAPK)signaling pathway.Twenty-five candidate genes for vitellogenesis and endocrine regulation were identified,such as vitellogenin,vitellogenin receptor,estrogen sulfotransferase,ecdysone receptor,prostaglandin reductase 1,hematopoietic prostaglandin D synthase and Juvenile hormone acid O-methyltransferase.And fifty-six genes related to nutrition metabolism were identified,such as fatty acid synthase,longchain-fatty-acid-Co A ligase 4,1-acyl-sn-glycerol-3-phosphate acyltransferase 4,fatty acid-binding protein and glycerol-3-phosphate acyltransferase 1.2.Comparative proteomics elucidates the dynamics of ovarian development in E.sinensisComparative proteomics analysis of the ovary and hepatopancreas at three key ovarian stages,including stages I(oogonium proliferation),II(endogenous vitellogenesis)and IV(exogenous vitellogenesis),of the Chinese mitten crab using a label-free quantitative approach.The results showed that a total of 2,224 proteins were identified,and some key proteins related to ovarian development and nutrition metabolism were differentially expressed.The 26 key proteins were mainly involved in the ubiquitin/proteasome pathway(UPP),cyclic AMP-protein kinase A(c AMP-PKA)signaling pathway,and mitogen-activated protein kinase(MAPK)signaling pathway during oogenesis.Fifteen differentially abundant proteins(DAPs)were found to participate in vitellogenesis and oocyte development,such as vitelline membrane outer layer protein 1 homolog,vitellogenin,vitellogenin receptor,heat shock 70 k Da protein cognate 3 and farnesyl pyrophosphate synthase.Forty-seven DAPs related to nutrition metabolism were identified,including the protein digestion,fatty acid metabolism,prostaglandin metabolism,lipid digestion and transportation,i.e.short-chain specific acyl-Co A dehydrogenase,acyl-Co A desaturase,fatty acid-binding protein,long-chain fatty acid Co A ligase 4,and hematopoietic prostaglandin D synthase.In conclusion,c AMP-PKA signaling pathway and ubiquitin protein degradation were particularly important during ovarian development in E.sinensis,the metabolism of arachidonic acid and prostaglandin were extremely important in the regulation of ovarian maturation.3.Using lipidomics to reveal details of lipid accumulation during ovarian development in E.sinensisIn order to further understand the metabolism of lipids during ovarian development in E.sinensis,the lipid changes in key tissues(ovary,hepatopancreas and serum)during ovarian development of E.sinensis were determined and compared by using lipidomics.The results showed that phosphatidylcholine,triglyceride and phosphatidylethanolamine were the most abundant lipid types in the ovary,hepatopancreas and serum of E.sinensis.In the ovary,the content of most triglycerides were increased during the ovarian development,with the highest relative content of TG(20:0/16:0/20:1)molecules,most phosphatidylcholines showed an upward trend,and PC(40:6)had the highest relative content,most phosphatidylethanolamines showed a downward trend,and PE(18:1e/20:5)had the highest relative content;In the hepatopancreas,the content of most triglycerides were the highest in stage III or IV,the relative content of TG(4:0/16:0/16:1)was the highest,most of the phosphatidylcholine showed a downward trend,and the relative content of PC(18:0e/18:1)was the highest;In serum,diglyceride showed an upward trend in the stage I-V,and the relative content of DG(18:2/20:4)was the highest,the results of phosphatidylcholines in serum were consistent with ovarian phosphatidylcholines.In conclusion,during the ovarian development of E.sinensis,the ovary mainly accumulated triglyceride,phosphatidylcholine and phosphatidylethanolamine,the contents of phosphatidylcholine and sphingomyelin significantly decreased in the hepatopancreas,and the main lipid molecule type was phosphatidylcholine in serum. |
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