异齿裂腹鱼Leptin和CART基因表达组织分布和摄食功能研究
|
摘要
本研究通过分析Leptin和CART基因在异齿裂腹鱼中的组织分布和摄食功能,为研究异齿裂腹鱼摄食调控机制提供理论依据。选取60尾体重相近体况健康的异齿裂腹鱼,随机分为2个试验处理,分别为摄食试验组和禁食复喂试验组。分别取异齿裂腹鱼脑、肝脏、心脏、肌肉、眼、肾脏、肠道、鳃、皮肤、鳔和脾脏,利用Realtime-PCR法检测Leptin和CART基因在异齿裂腹鱼不同组织中的表达水平以及摄食、禁食复喂对两种基因表达水平的影响。结果表明,异齿裂腹鱼Leptin和CART基因广泛的分布于各个组织中,其中Leptin基因在肝脏组织中表达量显著高于其他组织,CART基因在肠道和脑组织中表达量显著高于其他组织。摄食后1 h和3 h,Leptin的表达量显著高于摄食前试验组(P<0.05),CART的表达量极显著低于摄食前试验组(P<0.01);禁食第1 d、3 d和5 d时,Leptin和CART基因的表达量极显著低于对照组(P<0.01);而复喂后,Leptin基因的表达量没有显著性变化(P>0.05),CART基因的表达量极显著(P<0.01)上升。结果表明,Leptin是异齿裂腹鱼的摄食后饱感信号因子,而CART基因不是,但Leptin和CART基因均参与异齿裂腹鱼的调控摄食。
In this paper, the tissue distribution and appetite function of Leptin and CART gene of Schizothorax o'connori in Tibet were researched. In order to provide the theoretical basis for studying the appetite regulation mechanism, 60 Schizothorax o'connori with similar body weight were randomly divided into two experimental treatments: feeding group, fasting and refeeding group. The Leptin and CART mRNA expressions in different tissues and the effect of feeding on the genes expression were analyzed by Real-time PCR technology. The results showed that Leptin and CART mRNA were widely distributed in different tissues. The expression level of Leptin gene in liver was significantly higher than that in other tissues. The expression of CART gene in intestinal and brain was significantly higher than that in other tissues. At 1 h and 3 h after feeding, the expression of Leptin was significantly higher than that of the pre-prandial test group(P<0.05), and the expression of CART was significantly lower than that of the pre-prandial test group(P<0.01). In fasting experiment, Leptin and CART gene expression were both decreased after fasting. Leptin gene expression had no significantly changed and CART gene expression was increased after refeeding. The results showed that Leptin was the post-prandial satiety signal factors in Schizothorax o'connori, while CART might not be. The genes of Leptin and CART all have a function in regulation of appetite.
In this paper, the tissue distribution and appetite function of Leptin and CART gene of Schizothorax o'connori in Tibet were researched. In order to provide the theoretical basis for studying the appetite regulation mechanism, 60 Schizothorax o'connori with similar body weight were randomly divided into two experimental treatments: feeding group, fasting and refeeding group. The Leptin and CART mRNA expressions in different tissues and the effect of feeding on the genes expression were analyzed by Real-time PCR technology. The results showed that Leptin and CART mRNA were widely distributed in different tissues. The expression level of Leptin gene in liver was significantly higher than that in other tissues. The expression of CART gene in intestinal and brain was significantly higher than that in other tissues. At 1 h and 3 h after feeding, the expression of Leptin was significantly higher than that of the pre-prandial test group(P<0.05), and the expression of CART was significantly lower than that of the pre-prandial test group(P<0.01). In fasting experiment, Leptin and CART gene expression were both decreased after fasting. Leptin gene expression had no significantly changed and CART gene expression was increased after refeeding. The results showed that Leptin was the post-prandial satiety signal factors in Schizothorax o'connori, while CART might not be. The genes of Leptin and CART all have a function in regulation of appetite.
引文
[1]西藏自治区水产局.西藏鱼类及其资源[M].北京:中国农业出版社, 1995.
[2]Valassi E, Scacchi M, Cavagnini F. Neuroendocrine control of food intake[J]. Nutrition, metabolism and cardiovascular diseases. 2008, 18(2):158-168.
[3]季强.异齿裂腹鱼食性的初步研究[J].水利渔业, 2008,28(3):51-53, 82.
[4]Yao J L, Chen Y F, Chen F, et al. Age and growth of an endemic Tibetan fish, Schizothorax o`connori, in the Yarlung Tsangpo River[J]. Journal of freshwater ecology, 2009, 24:343-345.
[5]张良松.异齿裂腹鱼人工规模化繁殖技术研究[J].淡水渔业, 2011, 41(5):88-91, 95.
[6]张良松.异齿裂腹鱼胚胎发育与仔鱼早期发育的研究[J].大连水产学报, 2011, 26(3):238-242.
[7]Murashita K, Uji S, Yamamoto T, et al. Production of recombinant leptin and its effects on food intake in rainbow trout(Oncorhynchud mykiss)[J]. Comparative Biochemistry and Physiology. Part B, Biochemistry&Molecular Biology, 2008, 150(4):377-384.
[8]Chu D L H, Li V W T, Yu R M K. Leptin:clue to poor appetite in oxygen-starved fish[J]. Molecular and Cellular Endocrinology, 2010, 319(1):143-146.
[9]Trombley S, Maugars G, Kling P, et al. Effects of long-term restricted feeding on plasma leptin, hepatic leptin expression and leptin receptor expression in juvenile Atlantic salmon(Salmo salar L.)[J]. General and Comparative Endocrinology, 2012, 175(1):92-99.
[10]Maixnerova J, Hlavacek J, Blokesova D, et al. Structureactivity relationship of CART(cocaine and amphetamineregulated transcript)peptide fragments[J]. Peptides, 2007,28(10):1945-1953.
[11]Volkoff H, Peter R E. Effects of CART peptides on food consumption, feeding and associated behaviors in the goldfish, Carassius auratus:actions on neuropeptide Y-and orexin A-induced feeding[J]. Brain Res, 2000, 887(1):125-133.
[12]陈锋.雅鲁藏布江外来鲫的生活史对策研究[D].中国科学院水生生物研究所, 2009.
[13]Bailleul B, Akerblom I, Strosberg A. The leptin receptor promoter controls expression of a second distinst protein[J].Nucleic Acids Research, 1997, 25:2752-2758.
[14]Procaccini C, Jirillo E, Matarese G. Leptin as an immunomodulator[J]. Molecular Aspects of Medicine,2011, 33:35-45.
[15]Volkoff H, Eykelbosh A J, Peter R E. Role of leptin in the control of feeding of goldfish(Carassius auratus):interactions with cholecystokinin, neuropeptide Y and orexin A, and modulation by fasting[J]. Brain Research,2003, 962(1/2):90-109.
[16]Gorissen M, Berrier N J, Nabuurs S B, et al. Two divergent leptin paralogues in zebrafish(Danio rerio)that originate early in teleostean evolution[J]. Journal of Endocrinology.2009, 201(3):329-339.
[17]Huising M O, Geven E J, Kruiswijk C P, et al. Increased leptin expression in common carp(Cyprinus carpio)after food intake but not after fasting or feeding to satiation[J].Endocrinology. 2006, 147(12):5786-5797.
[18]Douglass J, Mckinzie A A, Couceyro P. PCR differential display identifies a rat brain mRNA that is transcriptionally regulated by cacaine and amphetamine[J]. Journal of Neuroscience, 1995, 15(3/2):2471-2481.
[19]Kehoe A S, Volkoff H. Cloning and characterization of neuropeptide Y(NPY)and cocaine and amphetamine regulated transcript(CART)in Atlantic cod(Gadus morhua)[J]:Comparative Biochemistry and Physiology Amolecualr, 2007, 146(3):451-461.
[20]Zhang H, Chen H, Zhang Y, et al. Molecular cloning,characterization and expression profiles of multiple leptin genes and a leptin receptor gene in orange-spotted grouper(Epinephelus coioides)[J]. General and comparative endocrinol, 2013, 181:295-305.
[21]Havel P J. Peripheral signals conveying metabolic information to the brain:short-term and long-term regulation of food intake and energy homeostasis[J].Experimental Biology and Medicine. 2001, 226(11):963-977.
[22]Valen R, Jordal A E, Murashita K, et al. Postprandial effects on appetite-related neuropeptide expression in the brain of Atlantic salmon, Salmo salar[J]. General and comparative endocrinology, 2011, 171(3):359-366.
[23]Peterson B C, Waldbieser G C, Riley J L, et al. Pre-and postprandial changes in orexigenic and anorexigenic factors in channel catfish(Ictalurus punctatus)[J]. General and Comparative Endocrinology, 2012, 176(2):231-239.
[24]Yuan D, Wer R, Wang T, et al. Appetite regulation in Schizothorax prenanti by three CART genes[J]. General and Comparative Endocrinology, 2015, 224:194-204.
[2]Valassi E, Scacchi M, Cavagnini F. Neuroendocrine control of food intake[J]. Nutrition, metabolism and cardiovascular diseases. 2008, 18(2):158-168.
[3]季强.异齿裂腹鱼食性的初步研究[J].水利渔业, 2008,28(3):51-53, 82.
[4]Yao J L, Chen Y F, Chen F, et al. Age and growth of an endemic Tibetan fish, Schizothorax o`connori, in the Yarlung Tsangpo River[J]. Journal of freshwater ecology, 2009, 24:343-345.
[5]张良松.异齿裂腹鱼人工规模化繁殖技术研究[J].淡水渔业, 2011, 41(5):88-91, 95.
[6]张良松.异齿裂腹鱼胚胎发育与仔鱼早期发育的研究[J].大连水产学报, 2011, 26(3):238-242.
[7]Murashita K, Uji S, Yamamoto T, et al. Production of recombinant leptin and its effects on food intake in rainbow trout(Oncorhynchud mykiss)[J]. Comparative Biochemistry and Physiology. Part B, Biochemistry&Molecular Biology, 2008, 150(4):377-384.
[8]Chu D L H, Li V W T, Yu R M K. Leptin:clue to poor appetite in oxygen-starved fish[J]. Molecular and Cellular Endocrinology, 2010, 319(1):143-146.
[9]Trombley S, Maugars G, Kling P, et al. Effects of long-term restricted feeding on plasma leptin, hepatic leptin expression and leptin receptor expression in juvenile Atlantic salmon(Salmo salar L.)[J]. General and Comparative Endocrinology, 2012, 175(1):92-99.
[10]Maixnerova J, Hlavacek J, Blokesova D, et al. Structureactivity relationship of CART(cocaine and amphetamineregulated transcript)peptide fragments[J]. Peptides, 2007,28(10):1945-1953.
[11]Volkoff H, Peter R E. Effects of CART peptides on food consumption, feeding and associated behaviors in the goldfish, Carassius auratus:actions on neuropeptide Y-and orexin A-induced feeding[J]. Brain Res, 2000, 887(1):125-133.
[12]陈锋.雅鲁藏布江外来鲫的生活史对策研究[D].中国科学院水生生物研究所, 2009.
[13]Bailleul B, Akerblom I, Strosberg A. The leptin receptor promoter controls expression of a second distinst protein[J].Nucleic Acids Research, 1997, 25:2752-2758.
[14]Procaccini C, Jirillo E, Matarese G. Leptin as an immunomodulator[J]. Molecular Aspects of Medicine,2011, 33:35-45.
[15]Volkoff H, Eykelbosh A J, Peter R E. Role of leptin in the control of feeding of goldfish(Carassius auratus):interactions with cholecystokinin, neuropeptide Y and orexin A, and modulation by fasting[J]. Brain Research,2003, 962(1/2):90-109.
[16]Gorissen M, Berrier N J, Nabuurs S B, et al. Two divergent leptin paralogues in zebrafish(Danio rerio)that originate early in teleostean evolution[J]. Journal of Endocrinology.2009, 201(3):329-339.
[17]Huising M O, Geven E J, Kruiswijk C P, et al. Increased leptin expression in common carp(Cyprinus carpio)after food intake but not after fasting or feeding to satiation[J].Endocrinology. 2006, 147(12):5786-5797.
[18]Douglass J, Mckinzie A A, Couceyro P. PCR differential display identifies a rat brain mRNA that is transcriptionally regulated by cacaine and amphetamine[J]. Journal of Neuroscience, 1995, 15(3/2):2471-2481.
[19]Kehoe A S, Volkoff H. Cloning and characterization of neuropeptide Y(NPY)and cocaine and amphetamine regulated transcript(CART)in Atlantic cod(Gadus morhua)[J]:Comparative Biochemistry and Physiology Amolecualr, 2007, 146(3):451-461.
[20]Zhang H, Chen H, Zhang Y, et al. Molecular cloning,characterization and expression profiles of multiple leptin genes and a leptin receptor gene in orange-spotted grouper(Epinephelus coioides)[J]. General and comparative endocrinol, 2013, 181:295-305.
[21]Havel P J. Peripheral signals conveying metabolic information to the brain:short-term and long-term regulation of food intake and energy homeostasis[J].Experimental Biology and Medicine. 2001, 226(11):963-977.
[22]Valen R, Jordal A E, Murashita K, et al. Postprandial effects on appetite-related neuropeptide expression in the brain of Atlantic salmon, Salmo salar[J]. General and comparative endocrinology, 2011, 171(3):359-366.
[23]Peterson B C, Waldbieser G C, Riley J L, et al. Pre-and postprandial changes in orexigenic and anorexigenic factors in channel catfish(Ictalurus punctatus)[J]. General and Comparative Endocrinology, 2012, 176(2):231-239.
[24]Yuan D, Wer R, Wang T, et al. Appetite regulation in Schizothorax prenanti by three CART genes[J]. General and Comparative Endocrinology, 2015, 224:194-204.