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1 38 6 Vol. 38, No ACTA HYDROBIOLOGICA SINICA Nov., doi: / 刘群温海深李吉方何峰 (, ) :, T3(FT3) T4(FT4) (TG) (TC), IGF- TR-α, FT3 FT4, 11 ; TG, TC,, 11 3(GR3) TG 1(GR1), 1 3 (P<0.05); IGF- TR-α FT3 FT4,, (SGR) TR-α, IGF-, IGF- : ; FT3; FT4; TG; IGF- ; TR-α 中图分类号 : S965.2 文献标识码 : A 文章编号 : (2014) , [1 3] [4],,,, [5], (GH) [6] (TH; T3 T4) - (IGF- ) [7, 8] IGF-,, [9] 1957 Salmon Daughaday (IGFs), [10] Mccomick (Oncorhynchus keta) IGF-, [11] ; Kagawa (Pagrus major)igf- [12] (TR), TR,, T3 [13] TR A-F6, A/B, C DNA, E F [14] ( ) [15], (Oncorhynchus mykiss) (Salmonidae), 1866, 收稿日期 : ; 修订日期 : 基金项目 : ( ) ( ) 作者简介 : (1985 ),, ; ; lq @163.com 通信作者 : (1963 ),, ; ; wenhaishen@ouc.edu.cn

05); IGF- TR-α FT3 FT4,, (SGR) TR-α, IGF-, IGF- : ; FT3; FT4; TG; IGF- ; TR-α 中图分类号 : S965.

2 6 : 1077,,,,,,, [16] T3(FT3) T4(FT4) (Triglyceride, TG) (Total cholesterin, TC), IGF- TR-α mrna,, IGF- TR-α, 1 1.1,,, 20, (3 m 3 m 3 m), 4.5 (GR1) 6.6 (GR2) 8.6 kg/m 3 (GR3), 3 8:00 17:00, 40% 26% 14% ph ( 1), 4, (MS-222, g/m 3 ), (±0.1 g),, 1.5 ml 4 6h, r/min 10min ( 125 I) (RIA) FT3 FT4, BS-180 TG TC, FT3 FT4, TG TC, ( ), RNA RNAiso Reagent Taq DNase (RNasefree) RNasin RNA SYBR Premix Ex Taq TaKaRa 1.3 PCR Primer5.0 IGF- (M ) TR-α(AF )cDNA ( 2), IGF- 176 bp, TR-α 140 bp Tab. 1 表 1 不同采样时间养殖水体指标的变化 Changes of water parameters at different sampling times Items 5 May 7 July 9 September 11 November 1 January of next year 3 March of next year ph T ( ) DO (mg/l) 表 2 虹鳟 IGF-Ⅰ TR-α 以及 18S 基因表达所用的引物 Tab. 2 Primers used for IGF-, TR-α and 18S of O. mykiss Primers IGF- F IGF- R TR-α F TR-α R 18S 18S (5-3 ) Sequence of the primers GTCCTAACCCTGACTTCG CAGCACTCGTCCACAATA AAACTGCCCATGTTCTCG GCCATCTCGCCGCTAA CCTGAGAAACGGCTACCACATC CCAATTA CAGGGCCTCGAAAG Fragment length 176 bp 140 bp 119 bp

2 FT3 FT4, TG TC, ( ), RNA RNAiso Reagent Taq DNase (RNasefree) RNasin RNA SYBR Premix Ex Taq TaKaRa 1.3 PCR Primer5.0 IGF- (M95183.1) TR-α(AF302245.1)cDNA ( 2), IGF- 176 bp, TR-α 140 bp Tab.

3 cdna 10, cdna PCR, 2 : SYBR Premix Ex Taq 10 μl 0.5 μl, 2 μl, ddh 2 O 20 μl : 95 1min; 95 10s, 60 (TR-α 62 ) 40s, 72 40s, 40, cdna,, PCR, 18S, 18S Ct, 2 ΔΔCt SPSS (One-Way ANOVA), Duncan s ±, P<0.05 Excel FT3 FT4 1 FT3 FT4, FT3 FT4, 11 GR1 GR3 (P<0.05); 11, FT3 FT4, 11 11,,, (SGR), SGR, 9 11, SGR 1 FT3 FT4 Fig. 1 Effects of different stocking density on the contents of FT3 and FT4 of O. mykiss, and changes of water temperature and Specific growth rate (SGR) (P<0.05); Different letters represent significant differences (P<0.05); the same applies bellow 2.2 TG TC 2,,, 2B, 11 GR3 TG GR1, 1

05); 11, FT3 FT4, 11 11,,, (SGR), SGR, 9 11, SGR 1 FT3 FT4 Fig. 1 Effects of different stocking density on the contents of FT3 and FT4 of O.

4 6 : 1079 (P<0.05) TC, (P>0.05) 2.3 IGF- TR-α RT-PCR IGF- TR-α 10 ( 3) IGF-, ; TR-α 10, IGF-, TR-α Fig. 2 2 TG TC Effects of different stocking density on the contents of TG and TC of O. mykiss 3 IGF- TR-α Fig. 3 The expression of IGF- and TR-α in O. mykiss HK. Head kidney; B. Brain; K. Kidney; SP. Spleen; L. Liver; MU. Muscle; G. Gill; PC. Pyloric caeca; H. Heart; I. Intestines; M. marker 2.4 IGF- TR-α 4A B, IGF-, 11, IGF- GR1 GR2 GR3 (P<0.05); GR1 IGF- GR3 (P<0.05) TR-α 4C D, 11, TR-α GR1 GR3 (P<0.05), GR2 GR3 GR1(P<0.05) 3, [17 20] [21, 22],, [23, 24] IGF IGF- IGF-, IGF-

3 The expression of IGF- and TR-α in O. mykiss HK. Head kidney; B. Brain; K. Kidney; SP. Spleen; L. Liver; MU. Muscle; G. Gill; PC.

5 (A) (B) IGF- mrna (C) (D) TR-α mrna Fig. 4 A and B indicate the effects of different stocking density on the mrna level of IGF- in liver (A)and the intestines of O. mykiss(b) and TR-α in liver(c) and kidney(d) - -, IGF- (GH), GH GH, [25], IGF- [26], Shepherd (Oreochromis mossambicus) GH, IGF- [27] IGF-, IGF- [28] [29] IGF- IGF-, (Cyprinus carpio) IGF-, [30] IGF-,, 11 IGF-, 9 10 SGR, T4, T3 T4, FT3 FT4, (Oreochromis niloticus) T3 T3, T3, T3, T3, [31] FT3 FT4,, SGR, Brown [32] Vijayan Leatherland (Salvelinus fontinalis), 120 kg/m 3 T3 T4 60 kg/m 3 T3 T4 [33]

mykiss(b) and TR-α in liver(c) and kidney(d) - -, IGF- (GH), GH GH, [25], IGF- [26], Shepherd (Oreochromis mossambicus) GH, IGF- [27] IGF-,

6 6 : 1081 (TR), TR TR-α TR-β TR (Scophthatmus maximus), [34],, TR-α, T3 TR-α, TR-α FT3 FT4, FT3 FT4, TR-α TG TC,,, :, 36h (Pseudosciaena crocea) TG TC [35],, [36] Di Marco (Dicentrarchus labrax) (15 45 kg/m 3 ) TC, [37], (FFA) [38] Leatherland FFA, TG, [39] TG TC, ,, TG TC,, 11 1 TG,,, TG, [40], TG TC,, IGF- [29] T3 IGF- [41], T3 IGF- IGF- FT3 FT4, [1] Vijayan M M, Ballantyne J S, Leatherland J F. High stocking density alters the energy metabolism of brook charr, Salvelinus fontinalis [J]. Aquaculture, 1990, 88(3 4): [2] Davisk B, Parker N C. Plasma corticosteroid stress response of fourteen species of warm water fish to transportation [J]. Transactions of the American Fisheries Society, 1986, 115(7): [3] Wang W B, Li A H. The effects of environmental stress to fish immune system [J]. Journal of Fisheries of China, 2002, 26(4): [,.., 2002, 26(4): ] [4] Jiang R D. Effects of stocking density on growth of rainbow trout juvenile [J]. Chinese Journal Fisheries, 2009, 22(4): [.., 2009, 22(4): 31 33] [5] Zhang T J, Yang Z C, Sun R Y. The acclimation of fishes to high stocking density [J]. Fisheries Science & Technology Information, 1998, 25(3): [,,.., 1998, 25(3): ] [6] Li W S, Lin H R, Wong A O. Effects of gonadotropin-releasing hormone on growth hormone secretion and gene expression in common carp pituitary [J]. Comparative Biochemistry and Physiology Part B, 2002, 132(2): [7] Cao Q P, Duguay S, Plisetskaya E, et al. Nucleotide sequence and growth hormone-regulated expression of salmon insulin-like growth factor I mrna [J]. Molecular and Cellular Endocrinology, 1989, 3(12): [8] Daughaday W, Rotwein P. Insulin-like growth factors I and II. Peptide, messenger ribonucleic acid and gene structures, serum, and tissue concentrations [J]. Endocrine Reviews, 1989, 10(1): [9] Clay L A, Wang S Y, Wolters W R, et al. Molecular characterization of the insulin-like growth factor-i(igf-i)

2,, TG TC,, 11 1 TG,,, TG, [40], TG TC,, IGF- [29] T3 IGF- [41], T3 IGF- IGF- FT3 FT4, [1] Vijayan M M, Ballantyne J S, Leatherland J F.

7 gene in channel catfish (Ictalurus punctatus) [J]. Biochimica et Biophysica Acta-Biomembranes, 2005, 1731(3): [10] Salmon W D, Daughaday W H. A hormonally controlled serum factor which stimulates sulfate in corporation by cartilage in vitro [J]. Journal of Laboratory and Clinical Medicine, 1957, 49(6): [11] Mccormick S D. Osmoregulatory action of insulin-like growth factor- in rainbow trout (Oncorhynchus mykiss) [J]. Endocrinology, 1991, 130(1): [12] Kagawa H. Immunocytochemical localization of IGF- in the ovary of the red sea bream, Pagrus major [J]. General and Comparative Endocrinology, 1995, 99(3): [13] Lee H J, Yen P M. Recent advances in understanding thyroid hormone receptor co-regulators [J]. Journal of Biomedical Science, 1999, 6(10): [14] Yen P M. Physiological and molecular basis of thyroid hormone action [J]. Physiological Reviews, 2001, 81(3): [15] Dong S L, Wang Z B, Lei X Q, et al. Influence of heat stress on biochemical indexes in blood of animal [J]. Ecology of Domestic Animal, 2004, 25(2): [,,,.., 2004, 25(2): 54 56] [16] Ellis T, North B, Scott A P, et al. The relationships between stocking density and welfare in farmed rainbow trout [J]. Journal of Fish Biology, 2002, 61(3): [17] Zhang Y Z, Liu D E, Fang Q S, et al. Effects of temperature and salinity on growth and survival rate of Young Epinephelus coioides [J]. Journal of Jimei University (Natural Science), 2009, 14(1): 8 13 [,,,.. ( ), 2009, 14(1): 8 13] [18] Jian Y X, Pan L, Hu F W, et al. The effects of temperature and salinity on survival and growth of larval Hexagrammos otakii Jordan et Starks [J]. Progress in Fishery Science, 2012, 33(5): [,,,.., 2012, 33(5):24 29] [19] Ruan C X, Wu D F, Yuan C G. Effects of temperature on the growth and hydrochemical state of juvenile Pseudosciaena crocea [J]. Journal of Guangzhou University (Natural Science Edition), 2013, 12(2): [,,. (Pseudosciaena crocea). ( ), 2013, 12(2): 36 39] [20] Guo Q D, Wang Y J, Lü W Q. Combined effects of temperature and salinity on the physiological osmotic induction and antioxidant responses in the juvenile Japanese flounder (Paralichthys olivaceus) [J]. Acta Hydrobiologica Sinica, 2014, 38(1): 58 67[,,.., 2014, 38(1): 58 67] [21] Wei K J, Zhang G R, Zhang H M. Studies on the phototactic characteristics of mandarin fish during different development stages [J]. Journal of Huazhong Agricultural University, 2001, 20(2): [,,.., 2001, 20(2): ] [22] Zhang Y Y, Bai Q L, Jia Z Y. Effects of light intensity on growth performance in juvenile Masu Salmon Oncorhynchus masou masou in Aquaculture [J]. Chinese Journal of Fisheries, 2012, 25(4): [,,.., 2012, 25(4): 33 35] [23] Allen K O. Effects of stocking density and water exchange rate on growth and survival of channel catfish Ictalurus punctatus in circular tanks [J]. Aquaculture, 1974, 4(null): [24] Andrews J W, Knight L H, Page J W. Interactions of stocking density and water turnover on growth and food conversion of channel catfish reared in intensively stocked tanks [J]. Progressive Fish-Culturist, 1971, 33(4): [25] Bolton J P, Collie N L, Kawauchi H, et al. Osmoregulatory actions of growth hormone in rainbow trout (Salmo gairdneri) [J]. Journal of Endocrinology, 1987, 112(2): [26] McCormick S D, Kelley K M, Young G, et al. Stimulation of coho salmon growth by insulin-like growth factor [J]. General and Comparative Endocrinology, 1992a, 86(3): [27] Shepherd B S, Sakamoto T, Nishioka R S, et al. Somatotropic actions of the homologous growth hormone (tgh) and prolactin (tprl177) in the euryhaline teleost, Oreochromis mossambicus [J]. Proceedings of the National Academy of Sciences of the United States of America, 1997, 94(5): [28] Zhang K D, Chen X C, Li Y W. Molecular cloning, sequence analysis and tissue expression of Insulin like growth factor in Chinese Sucker, Myxocyprinus asiaticus [J]. Chinese Journal of Zoology, 2007, 42(2): [,,. - (IGF- )cdna., 2007, 42(2): 39 45] [29] Zhang J L, Shi Z Y, Fu Y S. Gene expression and thyroid hormone regulated transcript of IGF-I during metamorphosis of the flounder, Paralichthys olivaceus [J]. Acta Hydrobiologica Sinica, 2011, 35(2): [,,. IGF-I., 2011, 35(2): ] [30] Hua Y M, Lin H R. Effects of different nutritional status on expression of IGF- mrna in immature common carp liver [J]. Acta Zoologica Sinica, 2001, 47(1): [,. IGF - m RNA., 2001, 47(1): ] [31] Ling M, Ye F L. Effects of salinity on serum Triiodothyronine and survival of Oreochromis niloticus [J]. Journal of Guangdong Ocean University, 2007, 27(3): [,.

Osmoregulatory action of insulin-like growth factor- in rainbow trout (Oncorhynchus mykiss) [J]. Endocrinology, 1991, 130(1): 87 92 [12] Kagawa H.

8 6 : 1083., 2007, 27(3): 33 37] [32] Brown C L, Doroshov S I, Nunes J M. Maternal triiodothyronine injections cause transitory elevations of plasma thyroxine in rainbow trout, Salmo garidneri [J]. Canadian Journal of Zoology, 1988, 56(1): [33] Vijayan M M, Leatherland J M. Effect of stocking density on the growth and stress-response in brook charr, Salvelinus fontinalis [J]. Aquaculture, 1988, 75(1): [34] Tian J, Shi Z Y. The expression of thyroid hormone receptor in the tissues and cells of Scophthatmus maximus and relationship between it and thyroid hormone [J]. Periodical of Ocean University of China, 2010, 40(Sup.): [,.., 2010, 40( ): ] [35] Ji D W, Li M Y, Wang T Z, et al. Effects of low temperature stress periods on serum biochemical indexes in large yellow coroaker Pseudosciaena crocea [J]. Fisheries Science, 2009, 28(1): 1 4 [,,,.., 2009, 28(1): 1 4] [36] Liu Z X, Tang X L, Zeng Y G. The mechanism of the influence from the heat stress on the chicken production [J]. Journal of Animal Science and Veterinary Medicine, 2001, 20(1): [,,.., 2001, 20(1): 29 32] [37] Di Marco P, Priori A, Finoia M G, et al. Physiological responses of European sea bass Dicentrarchus labrax to different stocking densities and acute stress challenge [J]. Aquaculture, 2008, 275(1 4): [38] Zhang L, Fan Q X, Zhao Z G, et al. The effects of chronic crowding stress on growth and blood biochemical indexes in common carp Cyprinus carpio [J]. Journal of Dalian Fisheries University, 2007, 22(6): [,,,.., 2007, 22(6): ] [39] Leatherland J F. Studies of the correlation between stress-response, osmoregulation and thyroid physiology in rainbow trout, Salmo gairdnerii (Richardson) [J]. Comparative Biochemistry and Physiology Part A: Physiology, 1985, 80(4): [40] Bo L, Qin Y W, Liu C P, et al. Prevalence of thyroid dysfunction in patients with dyslipidemia [J]. Chinese Journal of Clinical Laboratory Science, 2008, 26(2): [,,,.., 2008, 26(2): ] [41] Schmid A C, Lutz I, Kloas W, et al. Thyorid hormone stimulates hepatic IGF-I mrna expression in a bony fish, in the tilapia Oreochromism ossambicus [J]. General and Comparative Endocrinology, 2003, 130(2): EFFECTS OF DIFFERENT STOCKING DENSITY ON THE LEVELS OF THYRIOID HORMONE AND PLASMA LIPIDS IN ONCORHYNCHUS MYKISS LIU Qun, WEN Hai-Shen, LI Ji-Fang and HE Feng (College of Fisheries, Ocean University of China, Qingdao , China) Abstract: The aims of this study were to evaluate the effects of stocking density of Oncorhynchus mykiss that were reared in net cages on free triiodothyronine (FT3), free thyroxine (FT4), triglyceride (TG) and total cholesterin (TC) levels, and the expression of IGF-I and TR-α. The results showed that FT3 and FT4 levels decreased gradually as the stocking density increased, and they were positively associated with the culture time before November. TG levels decreased as the stocking density increased while TC levels increased as the stocking density increased and both TG and TC levels were negatively associated with the culture temperature. Moreover, TG levels in the treatment group 3(GR3) were significantly lower than those in the treatment group 1(GR1) in November, and significant differences were observed among the three treatments in January of the following year (P < 0.05). Additionally, the expression levels of IGF-I and TR-α decreased gradually as the stocking density increased. These results indicate that high stocking density could be a stressor for O. mykiss, that nd that may reduce FT3 and FT4 levels in the blood and then affect the growth and development of fish, which is supported by the trend of special growth rate (SGR). There was a relationship between thyroid hormone and the expression of IGF-I, The levels of thyroid hormone was related to the expression of IGF-I; however further investigation is needed to study whether thyroid hormone regulates the expression. Key words: Oncorhynchus mykiss; FT3; FT4; TG; IGF- ; TR-α

Aquaculture, 1988, 75(1):159 170 [34] Tian J, Shi Z Y. The expression of thyroid hormone receptor in the tissues and cells of Scophthatmus maximus and relationship between it and thyroid hormone [J].

22 Animal Husbandry & Veterinary Medicine 2011 Vol. 43 No % 3% 3% /% % 6. 67% 1. 71% 1. 11% 0. 14% 0.

22 Animal Husbandry & Veterinary Medicine 2011 Vol. 43 No % 3% 3% /% % 6. 67% 1. 71% 1. 11% 0. 14% 0. 2011 43 5 21 3 1 1* 1 2 3 1 1. 2. 3. 78. 68±2. 95 g 28 300 3 5 3% 3 1 ALT TP ALB AST 0. 05 P>0. 05 ALT TP ALB P>0. 05 AST 0. 05 GLOB P>0. 05 2 INS 0. 05 TSH T 3 0. 05 P>0. 05 GLU GH T4 IGF-I P>0. 05 S816.

Microsoft Word 刘 群_new_.doc

Microsoft Word 刘群_new_.doc