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Strategic Study of CAE >> 2014, Volume 16, Issue 9

cDNA Cloning of Neuropeptide Y gene of turbot Scophthalmus maximus L. and its expression analysis of the strategy of starvation and feeding in industrial culture

1. Key Laboratory of Mariculture & Stock Enhancement in North China’s Sea,Ministry of Agriculture,Dalian Ocean University,Dalian,Liaoning 116023,China;

2.Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology,Key Laboratory of Sustainable Development of Marine Fisheries,Ministry of Agriculture, Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao,Shandong 266071, China;

3. State Key Laboratory of Reproductive Biology Institute of Zoology,Chinese Academy of Sciences, Beijing 100101,China

Funding project:国家自然科学基金资助项目(30901111);山东省优秀中青年科学家奖励基金项目(BS2011HZ025);现代农业产业技术体系建设专项资金(CARS-50) Received: 2014-06-30 Available online: 2014-09-16 08:55:10.000

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Abstract

In this study, the full length of turbot NPY cDNA was cloned using RT-PCR and rapid-amplification of cDNA ends (RACE). Turbot NPY cDNA is 729bp with a 5´-untranslated region (UTR), 135bp 3´-UTR and 299bp ORF. The open reading frame encodes a 99 amino acid NPY. The first 28 amino acids constitute the signal sequence followed by the 36 amino acid mature peptide. Turbot NPY share a 53.9–98.0% identity with NPY from other fish, especially, Turbot NPY share a 97–98.0% identity with NPY from other flounder (Japanese flounder, winter flounder).Using real-time PCR to analysis NPY expression in turbot. Tissue distribution studies show that NPY mRNA is expressed in brain, in particular the hypothalamic, and in peripheral tissues. During food deprivation, the NPY undergo significant change in expression (P<0.05), with NPY levels being elevated first, then decreased, and being significantly increased, maintain a high level of expression, then decreased again during food deprivation. Expression level was significantly higher with refeeding after fasting than the normal feeding, the longer of food deprivation, the higher of expression level. But the expression level of 2h after refeeding will fall again to a similar level. Our results suggest that NPY is conserved peptides that might be involved in the regulation of feeding and other physiological functions in turbot.

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References

[ 1 ] Larhammar D,Wraith A,Berglund M M,et al. Origins of the many NPY-family receptors in mammals [J]. Peptides,2001,22 (3):295-307. link1

[ 2 ] 胡春燕,李英文. 神经肽 Y(NPY)的生理功能研究进展[J]. 生 物学杂志,2011,28(2):66-68. link1

[ 3 ] Tatemoto K. Neuropeptide Y:Complete amino acid sequence of the brain peptide [J]. Proceedings of the National Academy of Sciences of the United States of America,1982,79(18):5485- 5489. link1

[ 4 ] Cerdá-Reverter J M,Martínez-Rodríguez G,Zanuy S,et al. Molecular evolution of the neuropeptide Y (NPY) family of peptides:Cloning of three NPY- related peptides from the sea bass (Dicentrarchus labrax) [J]. Regulatory Peptides,2000,95(1): 25-34. link1

[ 5 ] Cerdá- Reverter José Miguel,Anglade Isabelle,Martı́nezRodrı́Guez Gonzalo,et al. Characterization of neuropeptide Y expression in the brain of a perciform fish,the sea bass (Dicentrarchus labrax) [J]. Journal of Chemical Neuroanatomy,2000,19(4):197-210. link1

[ 6 ] Dumont Y,Martel J C,Fournier A,et al. Neuropeptide Y and neuropeptide Y receptor subtypes in brain and peripheral tissues [J]. Progress in Neurobiology,1992,38(2):125-167. link1

[ 7 ] Peng C,Gallin W,Peter R E,et al. Neuropeptide- Y gene expression in the goldfish brain:Distribution and regulation by ovarian steroids [J]. Endocrinology,1994,134(3):1095-1103. link1

[ 8 ] Preston E,Jönsson A C,McManus C D,et al. Comparative vascular responses in elasmobranchs to different structures of neuropeptide Y and peptide YY [J]. Regulatory Peptides,1998,78(1- 3):57-67. link1

[ 9 ] Shahbazi F,Holmgren S,Larhammar D,et al. Neuropeptide Y effects on vasorelaxation and intestinal contraction in the Atlantic cod Gadus morhua [J]. American Journal of Physiology. Regulatory,Integrative and Comparative Physiology,2002,282(5): R1414-R1421. link1

[10] Volkoff H,Peter R E. Characterization of two forms of cocaineand amphetamine- regulated transcript (CART) peptide precursors in goldfish:Molecular cloning and distribution,modulation of expression by nutritional status,and interactions with leptin [J]. Endocrinology,2001,142(12):5076-5088. link1

[11] Doyon C,Gilmour K M,Trudeau V L,et al. Corticotropinreleasing factor and neuropeptide Y mRNA levels are elevated in the preoptic area of socially subordinate rainbow trout [J]. General and Comparative Endocrinology,2003,133(2):260- 271. link1

[12] Doyon Christian,Leclair Jason,Trudeau Vance L,et al. Corticotropin- releasing factor and neuropeptide Y mRNA levels are modified by glucocorticoids in rainbow trout,Oncorhynchus mykiss [J]. General and Comparative Endocrinology,2006,146 (2):126-135. link1

[13] Clark J T,Kalra P S,Crowley W R,et al. Neuropeptide Y and human pancreatic polypeptide stimulate feeding behavior in rats [J]. Endocrinology,1984,115(1):427-429. link1

[14] Boswell T,Dunn I C,Corr S A. Hypothalamic neuropeptide Y mRNA is increased after feed restriction in growing broilers [J]. Poultry Science,1999,78(8):1203-1207. link1

[15] Schwippert W W,Ewert J P. Effect of neuropeptide-Y on tectal field potentials in the toad [J]. Brain Research,1995,669(1): 150-152. link1

[16] Silverstein Jeffrey T,Plisetskaya Erika M. The effects of NPY and insulin on food intake regulation in fish [J]. American Zoologist,2000,40(2):296-308. link1

[17] Kah O,Pontet A,Danger J M,et al. Characterization,cerebral distribution and gonadotropin release activity of neuropeptide Y (NPY) in the goldfish [J]. Fish Physiology and Biochemistry,1989,7(1-6):69-76. link1

[18] Breton B,Mikolajczyk T,Danger J M,et al. Neuropeptide Y (NPY) modulatesin vitro gonadotropin in release from rainbow trout pituitary glands [J]. Fish Physiology and Biochemistry, 1989,7(1-6):77-83. link1

[19] Peng C,Chang J P,Yu K L,et al. Neuropeptide-Y stimulates growth hormone and gonadotropin-Ⅱ secretion in the goldfish pituitary:Involvement of both presynaptic and pituitary cell actions [J]. Endocrinology,1993,132(4):1820-1829. link1

[20] Peng C,Humphries S,Peter R E,et al. Actions of goldfish neuropeptide Y on the secretion of growth hormone and gonadotropin-Ⅱ in female goldfish [J]. General and Comparative Endocrinology,1993,90(3):306-317. link1

[21] Silverstein J T,Breininger J,Baskin D G,et al. Neuropeptide Y-like gene expression in the salmon brain increases with fasting [J]. General and Comparative Endocrinology,1998,110 (2):157-165. link1

[22] López-Patiño M A,Guijarro A I,Isorna E,et al. Neuropeptide Y has a stimulatory action on feeding behavior in goldfish (Carassius auratus) [J]. European Journal of Pharmacology, 1999,377(2/3):147-153. link1

[23] MacDonald Erin,Volkoff Hélène. Cloning,distribution and effects of season and nutritional status on the expression of neuropeptide Y (NPY),cocaine and amphetamine regulated transcript (CART) and cholecystokinin (CCK) in winter flounder (Pseudopleuronectes americanus) [J]. Hormones and Behavior, 2009,56(1):58-65. link1

[24] Kehoe Amy S,Volkoff Hélène. Cloning and characterization of neuropeptide Y (NPY) and cocaine and amphetamine regulated transcript (CART) in Atlantic cod (Gadus morhua) [J]. Comparative Biochemistry and Physiology Part A:Molecular & Integrative Physiology,2007,146(3):451-461. link1

[25] 雷霁霖,刘新富. 大菱鲆(Scophthalmus maximus L.)引进养殖 的初步研究[J]. 现代渔业信息,1995,10(11):1-3. link1

[26] 赵 玫,袁锦虹,李 嘉,等. 针刺对实验性肥胖大鼠下丘脑 摄食中枢的影响 [J]. 中国针灸,2001,21(5):305-307. link1

[27] Bjenning C,Hazon N,Balasubramaniam A,et al. Distribution and activity of dogfish NPY and peptide YY in the cardiovascular system of the common dogfish [J]. American Journal of Physiology. Regulatory,Integrative and Comparative Physiology,1993,264(6):R1119-R1124. link1

[28] Karila P,Messenger J,Holmgren S. Nitric oxide synthase- and neuropeptide Y- containing subpopulations of sympathetic neurons in the coeliac ganglion of the Atlantic cod, Gadus morhua,revealed by immunohistochemistry and retrograde tracing from the stomach [J]. Journal of the Autonomic Nervous System,1997,66(1/2):35-45. link1

[29] Gómez- Visus I,Garcı́a-Hernández M P,Lozano M T,et al. Glucagon- and NPY-related peptide-immunoreactive cells in the gut of sea bass (Dicentrarchus labrax L.):A light and electron microscopic study [J]. General and Comparative Endocrinology,1998,112(1):26-37. link1

[30] 陈文波,王 鑫,周雅璐,等. 尼罗罗非鱼Orexin前体基因的 克隆、组织分布及其在摄食调控中的表达[J]. 动物学研究, 2011,32(3):285-292. link1

[31] Volkoff H,Canosa L F,Unniappan S,et al. Neuropeptides and the control of food intake in fish [J]. General and Comparative Endocrinology,2005,142(1/2):3-19. link1

[32] Unniappan S,Canosa L F,Peter R E,et al. Orexigenic actions of ghrelin in goldfish:Feeding- induced changes in brain and gut mRNA expression and serum levels,and responses to central and peripheral injections [J]. Neuroendocrinology,2004,79 (2):100-108. link1

[33] Volkoff Hélène,Peter Richard Ector. 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 Research,2000,887(1): 125-133. link1

[34] Arends R J,Vermeer H,Martens G J,et al. Cloning and expression of two proopiomelanocortin mRNAs in the common carp (Cyprinus carpio L.) [J]. Molecular and Cellular Endocrinology,1998,143(1/2):23-31. link1

[35] Loh Y P,Gritsch H A,Chang T L. Pro- opiocortin processing in the pituitary:A model for neuropeptide biosynthesis [J]. Peptides,1982,3(3):397-404. link1

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