C1q/TNF-Related Protein 9 in the Orange-Spotted Grouper :Molecular Cloning, Identification and Its Function
Guokun Yang, Chaobin Qin, Qinqin Song, Caiyun Sun, WenSheng Li *
State Key Laboratory of Biocontrol, Institute of Aquatic Economic Animals and Guangdong Province Key Laboratory for Aquatic Economic Animals, Guangdong Provincial Engineering Technology Research Center of Healthy Breeding in Important Economic Fish, South China Sea Bio-Resource Exploitation and Collaborative Innovation Center, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China
Adipose tissue is not only an organ of energy storage, but also an active endocrine organ. It can release a variety of cytokines, e.g.adiponectin. Adiponectin is an important adipocytokines and has been studied intensively, since adiponectin can improve sensitivity of insulin and also regulate metabolism of glucose and lipid. CTRP9 is newly identified adipokine and has the similar biological function as adiponectin. In the present study, we cloned the full-length cDNA of CTRP9 from orange-spotted grouper, Epinephelus coioides. Using real-time PCR, ctrp-9 showed relative higher expression in the kidney and brain region at normal status, while at fasting time, the expression of ctrp-9 in the liver decreased significantly compared with the normal feeding group, it will recover after refeeding. But the expression of ctrp-9 in the adipose tissue did not show obvious change during fasting or refeeding. Our suggests that ctrp-9 has different expression mode at different energy status and tissues. Following successful production of recombinant CTRP9 (rgCTRP9) as well as generation of polyclone antibody of CTRP9, we have conducted pull-down assay and identified the endogenous expression of CTRP9 protein in orange-spotted grouper. The mRNA levels of adipose differentiation-related genes were affected in the adipose tissue after intraperitoneal injection of rgCTRP9. Similar observations were also found in the hypothalamus regarding the food intake-related genes. The role of CTRP9 on glucose and lipid metabolism were further examined in the primary hepatocytes culture and adipocyte culture. Interestingly, for the first time, we observed that rgCTRP9 might regulate reproduction-related genes in the hypothalamus, pituitary and mature gonad. Elucidating underlie molecular mechanisms of CTRP9 that mediate glucolipid metabolism and reproduction in teleosts requires additional studies.
Acknowledgements: Supported by National Science Foundation of China (31272639), Guangdong Provincial Science and Technology Program (2015A020216006), Guangdong Oceanic and Fishery Program (A201601C02), and Fundamental Research Funds for the Central Universities17lgjc18 to WL .
Key Words: CTRP9, clone, metabolism, reproduction, Epinephelus coioides
Guokun Yang, Chaobin Qin, Qinqin Song, Caiyun Sun, WenSheng Li *
State Key Laboratory of Biocontrol, Institute of Aquatic Economic Animals and Guangdong Province Key Laboratory for Aquatic Economic Animals, Guangdong Provincial Engineering Technology Research Center of Healthy Breeding in Important Economic Fish, South China Sea Bio-Resource Exploitation and Collaborative Innovation Center, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China
Adipose tissue is not only an organ of energy storage, but also an active endocrine organ. It can release a variety of cytokines, e.g.adiponectin. Adiponectin is an important adipocytokines and has been studied intensively, since adiponectin can improve sensitivity of insulin and also regulate metabolism of glucose and lipid. CTRP9 is newly identified adipokine and has the similar biological function as adiponectin. In the present study, we cloned the full-length cDNA of CTRP9 from orange-spotted grouper, Epinephelus coioides. Using real-time PCR, ctrp-9 showed relative higher expression in the kidney and brain region at normal status, while at fasting time, the expression of ctrp-9 in the liver decreased significantly compared with the normal feeding group, it will recover after refeeding. But the expression of ctrp-9 in the adipose tissue did not show obvious change during fasting or refeeding. Our suggests that ctrp-9 has different expression mode at different energy status and tissues. Following successful production of recombinant CTRP9 (rgCTRP9) as well as generation of polyclone antibody of CTRP9, we have conducted pull-down assay and identified the endogenous expression of CTRP9 protein in orange-spotted grouper. The mRNA levels of adipose differentiation-related genes were affected in the adipose tissue after intraperitoneal injection of rgCTRP9. Similar observations were also found in the hypothalamus regarding the food intake-related genes. The role of CTRP9 on glucose and lipid metabolism were further examined in the primary hepatocytes culture and adipocyte culture. Interestingly, for the first time, we observed that rgCTRP9 might regulate reproduction-related genes in the hypothalamus, pituitary and mature gonad. Elucidating underlie molecular mechanisms of CTRP9 that mediate glucolipid metabolism and reproduction in teleosts requires additional studies.
Acknowledgements: Supported by National Science Foundation of China (31272639), Guangdong Provincial Science and Technology Program (2015A020216006), Guangdong Oceanic and Fishery Program (A201601C02), and Fundamental Research Funds for the Central Universities17lgjc18 to WL .
Key Words: CTRP9, clone, metabolism, reproduction, Epinephelus coioides