Microsoft Word - 4柳学周_new_.doc

Size: px

Start display at page:

Download "Microsoft Word - 4柳学周_new_.doc"

釜元
7 years ago
Views:

1 35 4 Vol.35, No PROGRESS IN FISHERY SCIENCES Aug., 2014 DOI: /yykxjz (Paralichthys lethostigma)lh β * 1 1, ( ) cdna (Rapid Amplification of cdna Ends, RACE) (Paralichthys lethostigma) (Luteinizing Hormone, LH) cdna LH mrna LH mrna (Enzyme-Linked Immuno Sorbent Assay, ELISA) LH (Estrodiol, E 2 ) LH cdna 597 bp (Open reading frame, ORF) 438 bp 145 LH mrna LH mrna IV VI LH mrna V LH mrna II V LH E 2 LH S961 文献标识码 A 文章编号 (2014) (HPG) (Folliclestimulating hormone, FSH) (LH) LH (Maturation-inducing hormone, MIH)( 17α, 20β- ) MIH (Kagawa et al 1998) LH α FSH (Thyrotropin, Thyroid-stimulating hormone, TSH) (Pierce et al 1981) LH (Oncorhyn chustschawytscha)(trinh et al, 1986) (Oncorhyn chusketa)(sekine et al, 1989) (Morone saxatilis)(hassin et al, 1995) (Anguilla japonica)(yoshiura et al, 1996) (Carassiu sauratus)(sohn et al 1998) (Chrysophrys major)(gen et al 2000) 30 LH Pleuronectiformes Bothidae Paralichthys 2001 * (CARS-50) 863 (2012AA10A413) liuxz@ysfri.ac.cn : , :

2 4 : (Paralichthys lethostigma)lh β ( 2007) Tubbs (2011) MIH(20β-S) Picha (2012) IGF-II LH RACE RT-PCR ELISA LH cdna ( cm g) ph mg/l 300% 500% lx 12 h 10 h 1.2 ( ZL ) MS-222(180 mg/l) 2 3 ml 4 6 h r/min 5 min 30 LH E 2 80 RNA Davidson 24 h 70% % (70% 100%) 7 μm (NIKON 90i) 50% 30 (1978) 1.4 LH RNAiso Plus(TaKaRa) RNA (Nanodrop ND2000) RNA RNA 1% SMART TM RACE cdna (Clontech) cdna, GenBank LH LHF LHR( 1) cdna LH PCR 25 μl 94 5 min (94 30 s s s) min PCR 1% PCR peasy-t3 Trans1-T1 LB 37 RACE LHGSP5 LHGSP3 LHNGSP5 LHNGSP3( 1) SMART TM RACE cdna (TaKaRa) LH cdna 5'-RACE 3'-RACE cdna cdna Smart RACE Advantage 2 PCR (Clontech) PCR PCR LHGSP5 LHGSP3 PCR s Primers LHF LHR LHGSP5 LHGSP3 LHNGSP5 LHNGSP3 LHF1 LHR1 18S F 18S R 表 1 实验所用引物及其序列 Tab.1 Primer sequences used for the PCR analysis in the present study Nucleotide sequence(5'-3') GTACCAGCATGTGTGCACGT GTYGGGCTGCAGGCTCT TGGACCCGACCGTCTCCTACCCTGTGG TGGAAGGTGCAGTCGGACGTGTTGAGT CACTCAACACGTCCGACTGCACCTTCC GCAGTAACAGCTCACGGCCACAGGGTAG GTACCAGCACGTGTGCACGT GTCTGGCTGCAGGCTCT GGTAACGGGGAATCAGGGTT TCGGGAGTGGGTAATTTGCG

3 s 16 Tm 0.5, 72 1 min s s s 28 PCR LHNGSP5 LHNGSP3 PCR PCR s, s 5 5 (94 30 s, s, s) 25 (94 30 s, s, s) 5 μl PCR 1% 1.5 LH RNA 1 μg RNA 1.4 cdna cdna LHF1 LHF2 ( 1) 18S rrna (18SF 18SR) PCR PCR 94 5 min (94 30 s s s) min PCR 1% LH mrna LH mrna 1.6 LH E 2 LH E 2 (ELISA) (CUSABIO ) OD LH E 2 R IU/L 1.0 pmol/l 15% 1.7 DAMBE( LH Chromas ( BLAST ( BLAST/) DNAStar ( LH Clustalx2.0.12( MEGA 4.1( LH mrna ImageJ2x ( ) (ANOVA) (SPSS v 16.0) P 0.05 P< ( 1a) II 10 ( 1b) ( 1c) 11 ( 1d) IV 12 Fig.1 1 The morphology of oocytes at different developmental phases a. 40 b. 200 c. 200 d. 200 e. 200 a. Oocyte of stage II 40 b. Oocyte of stage III 200 c. Oocyte of stage IV 200 d. Oocyte of stage 200 e. Oocyte of stage VI 200

4 4 : (Paralichthys lethostigma)lh β 25 V 2 ( 1e) VI LH LH cdna (GenBank JX002656)( 2) 597 bp bp (ORF) 72 bp 5 87 bp 3 (TGA) DNAstar LH Da 5.58 LH N- C LH (Paralichthys olivaceus) 97.9% 74.5% 48.5% 51.8% (Gallus gallus) 37.9% (Equus caballus) 35.2%( 3) Neighbor-joining LH ( 4) LH 鮋 2.3 LH LH ( 5) 2.4 LH PCR LH mrna LH mrna 7 (II ) 11 (IV) VI ( 6) LH mrna II V V VI -108 ag tcc gtc tcc -60 ggc tca acc acc agc agc tgt tga atc gat cgt gga cac aaa cac ttc atc tcc tgc agg 1 ATG TCG ATG CAG CTC AGC CTG AGA GTG AAG ATC CCT CTG ACG TTG ATT TTC TTT CTC TCG M S M Q L S L R V K I P L T L I F F L S 61 TCC ATG TGG CCC CTG GCC CCC GCA GAG GCC TTC CAG CTA CCG ACG TGT CTT CTC ATC AAG S M W P L A P A E A F Q L P T C L L I K 121 CAA ATG GTG TCT CTG GAG AAG GAG GGC TGT CCC AAG TGT CAC ACC GTG GAA ACC ACC ATC Q M V S L E K E G C P K C H T V E T T I 181 TGC AGT GGA CAC TGC AAA ACC AAG GAT CCC GTC GCC AAG ATC CCC TTC CTC AAC ATG TAC C S G H C K T K D P V A K I P F L N M Y 241 CAG CAC GTG TGC ACG TAC CAG GAG CTG TAC TAC AAG ACG TTT GAA CTT CCA GAC TGT CCT Q H V C T Y Q E L Y Y K T F E L P D C P 301 CCA GGC GTG GAC CCG ACC GTC TCC TAC CCT GTG GCC GTG AGC TGT TAC TGC GGC CGC TGT P G V D P T V S Y P V A V S C Y C G R C 361 GCA CTC AAC ACG TCC GAC TGC ACC TTC CAG AGC CTG CAG CCA GAC TTC TGC ATG AAC GAC A L N T S D C T F Q S L Q P D F C M N D 421 ATC CCC TTC TAC GAC TGA acg tta aac act gga gtc caa acc atg ttt aat aaa aga cag I P F Y D * 481 atc ttt aca ttc acc act gcaaaaaaaaaaaaaaaaaaaaaaa Fig.2 2 LH cdna cdna sequence and induced amino acid sequence of LH β subunit gene (TGA) 3 -UTR aataa N- NTSD The asterisk indicates the stop codon (TGA). The nucleotides corresponding to the polyadenylation signal in the 3 -untranslated region (aataa) are marked with rims. The potential N-glycosylation site (NTSD) in N-region was single underlined

5 26 35 Southern flounder Japanese flounder Atlantic halibut Korean rockfish Longtooth grouper Striped sea-bass Swamp eel Senegalese sole Coho salmon Atlantic salmon Rainbow trout Russian sturgeon Marbled eel Zebrafish Grass carp Chicken Horse 1 -MSMQLSLRVKIPLTLIFFL-----SSMWPLAPAEAFQ------LPTCLLIKQMVSLEKE I...V V METE.I.V...L V K.Q MM-.AMF.VM.SWI.GAS--Y.I...A S.Q..N.T... 1 M.AV.VG-..MF..M.SL..GAS--..I.S...A P.Q..N.T AV.A.-..MF..V.SL..GAT--.DI P.Q..N.T AA.V.-..MFL.MIDL..GAS--..I...A P.Q..N.T... 1 MAAV.IR-.LTFH...TVL.GSS--T.D..P...VSS SP.Q.VN.T...D VG--TL.S.F.CIL E.M.GSL------MQP.QP.N.T... 1 MLGLHVG--TL.S.F.CIL E.V.GSL------MQP.QP.N.T... 1 MLGLHVG--TL.S.L.CIL E.V.GSL------MQP.QP.N.T..V MPASVS.LLL.F SA.VLSRSSS RL.EPVNETI.A MSVYPECTWL.FV---CLCHLPVS.GGSL LP.EP.NETI.V..D MLLAGNGV...F----.LFFL..A.QSLV------F.R.E.VNET..V MLVVRNILLL.F----CLVVL.VF.QSSF P.EPVNET.AV... 1 MGGA.VLVLMTLLG.PPATTGNPPVAVDP...VVGPPMGLGGGGR.P.RP.NVT.AV..D METLQGL.LWM.LS----VGGVW.SRGPLR L.RP.NATLAA... Southern flounder 49 GCPKCHTVETTICSGHCKTKDPVAKIPFLNMYQHVCTYQELYYKTFELPDCPPGVDPTVS Japanese flounder Atlantic halibut 50...N...I...V... Korean rockfish 47..S...P...P...I...S.V...DF...S...T Longtooth grouper 52...P...I...I...S.V.E...RDF.H...T Striped sea-bass 51...P...I...I...S.V...RD.H...T Swamp eel 51...P...I...I...S.V...RDW...F... Senegalese sole 52..HT..P...Y...I...DVH...H...G...S.T Coho salmon 37...T.LVIQ.P...V..E..F.S..STV...RDVR.E.IR...W...H.T Atlantic salmon 42...T.LVIQ.P...V..E..F.S..STV...RDVR.E.IR...W...H.T Rainbow trout 42...T.LVIQ.P...V..E..F.S..STV...RDVR.E.IR...W...H.T Russian sturgeon 39 E...LLIQ.S...S.P...F.SALSTVQ...KD.RFV.VT...HFT Marbled eel 43...LVFQ.S...I...SY.S.LSTV..R...RDVR.E.VR...R...H.T Zebrafish 42...LVFQ...V.R...Y.S..STVH.T...RDVR.E.IN...SA...QIT Grass carp 42...LVFQ...L..E..Y.S..STV...RDVR.E.VR...HIT Chicken 61...Q.MA.T..A.G.Y.R.RE..YRS.LGPPP.SA...GA.R.ERWA.WG..I.S..R.L Horse 42 A..I.I.FT.S..A.Y.PSMVR.MPAALPAIP.P...R..RFASIR..G...M.. Southern flounder 109 YPVAVSCYCGRCALNTSDCTFQSLQPDFCMNDIPFYD Japanese flounder Atlantic halibut E Korean rockfish L..H...MD...E...N...HY Longtooth grouper L..H...MD...E...N...Y Striped sea-bass Q..H...MD...E...N...Y Swamp eel L..H...D...YE..R...Y Senegalese sole S.S...MDM...YE...I...L..Y Coho salmon 97...L..D.SL.NMD...IE...ITQRVLT.GDMW Atlantic salmon L..D.SL.NMD...IE...ITHRALM.GNMW Rainbow trout L..D.SL.NMD...IE...ITQRVLT.GDMW Russian sturgeon 99 F.L.L..E.SL.RMES...I..VG.SD..SGELAIQNY Marbled eel 103 F...L..D.NL.TMD...AI...R...SQRASLPA Zebrafish L..D.SL.TI...I...SQREDFSAY Grass carp L..D.SL.TMD...IE...SQREDFPVY Chicken 121 L...L..R.A..PMA...V.G.G.A..GAPGG.GGE Horse 102 F...L..H..P.QIK.T..GVFRD..LA.APQASSSSKDPPSQPLTSTSTPTPGASRRSS Southern flounder Japanese flounder Atlantic halibut Korean rockfish Longtooth grouper Striped sea-bass Swamp eel Senegalese sole Coho salmon Atlantic salmon Rainbow trout Russian sturgeon Marbled eel Zebrafish Grass carp Chicken Horse 162 HPLPIKTS 3 LH β Fig.3 Comparison of LH β subunit precursors between southern flounder and other vertebrates

6 4 : (Paralichthys lethostigma)lh β 27 5 LH Fig.5 Expression pattern of LH β subunit in various tissues of Paralichthys lethostigma P B O L H K SP ST I M G P: pituitary; B: brain; O: ovary; L: liver; H: heart; K: kidney; SP: spleen; ST: stomach; I: intestine; M: muscle; G: gill 4 LH LH Fig.4 Unrooted phylogenetic tree of LH β in vertebrates using Neighbor-Joining method (Mega 4.1 software) according to the amino acid sequence alignments of Paralichthys lethostigma with ClustalX. Boot strap values are based on 1000 repeats. GenBank accession numbers: Sebastes schlegelii(aau14142), Epinephelusbruneus (ABQ57400), Morone saxatilis (AAC38019), Monoperus albus (ACF70665), Hippoglossus hippoglossus (CAD10502), Paralichthys olivaceus (AAK58602), Solea senegalensis (ABU95600), Oncorhyn chuskisutch (AAO72300), Oncorhyn chusmykiss (NP ), Salmo salar (NP ), Acipenser gueldenstaedtii (AAP97490), Anguilla anguilla(ack87153), Ctenopharyngodon idella (ABM73670), Danio rerio (AAV31153), Equus caballus (P08751), Gallus gallus (ADY03193) Paralichthys lethostigma is underlined ( 6) LH mrna II V ( 6) 6 LH Fig.6 Expression patterns of pituitary, ovary and liver LH β mrna levels in P. lethostigma during ovarian maturation cycle M: Marker; P: ; O: ; L: M: Marker; P: pituitary; O: ovary; L: liver 2.5 LH E 2 LH E 2

7 28 35 LH II III IV V VI ( 7) E 2 LH ( 8) 3 RACE RT-PCR LH LH E 2 (Verasper variegatus)( 2011) (Cynoglossus semilaevis)( 2009) - ( 2007) LH N- LH LH (Swanson et al, 2003) LH N- (Miranda et al, 2007) LH (Kajimura et al, 2001) LH mrna (Jackson et al, 1999; 7 LH ( ) Fig.7 Changes of plasma LH levels in female P. lethostigma during ovarian maturation cycle * P=0.05 (P < 0.05), n=5 *Different letters indicate significant difference at P=0.05 level (P < 0.05), n=5 8 E 2 ( ) Fig.8 Changes of plasma E 2 levels in female P. lethostigma during ovarian maturation cycle * P=0.05 (P < 0.05), n=5 *Different letters indicate significant differences at P=0.05 level (P < 0.05), n=5

8 4 : (Paralichthys lethostigma)lh β 29 Gen et al, 2000; Meiri et al, 2004; Hellqvist et al, 2006) LH LH (Danio rerio) LH mrna (So et al, 2005) LH Hellqvist (2006) LH LH LH mrna 3 (1) LH (Kobayashi et al, 1997) (Shimon et al, 1999) (Trichogaster trichopterus)(jackson et al, 1999) (2) LH (Gen et al, 2000) (Gasterosteus aculeatus)(hellqvist et al, 2006) (3) LH (Weil et al, 1995) (Meiri et al, 2004) LH mrna IV VI (Kobayashi et al, 1997) (Shimon et al, 1999) (Jackson et al, 1999) LH mrna LH ( ) LH mrna LH (Hellqvist et al, 2006) LH mrna ( ) LH LH MIH( 17, 20 - ) MIH (Kagawa et al, 1998) LH mrna ( ) LH LH LH mrna LH LH LH LH mrna LH LH LH mrna (Jackson et al, 1999) (Weil et al, 1995) (Shimon et al, 1999) Halban (1994) LH LH (II ) LH mrna VI LH mrna LH Gomez (1999) LH LH ( ) (IV ) LH LH mrna LH mrna LH LH LH (IV ) (Jackson et al, 1999) (Gen et al, 2000) (Oreochromis niloticus) (Aizen et al, 2007) LH LH (V ) LH LH mrna (VI ) LH LH mrna VI LH mrna (Scomber japonicus) LH (Nyuji et al, 2011) II LH E 2 LH

9 30 35 (Hirohiko et al, 2003) Hirohiko (2003) LH P450 E 2 Ando (2003) E 2 (Gonadotropin-releasing hormone, GnRH) LH LH E 2 E 2 II E 2 (2010) II E 2 ERa ( IV ) E 2 II E 2 (Lange et al, 2003) (V )E 2 (Methyen et al, 1992) (VI )E 2,,,.., 1978, 6(3): ,,,.., 2011, 18(4): ,,,.., 2010, 40(7): 66 72,,,.., 2007, 34(2): 14 17,,,.., 2009, 33(6): Aizen J, Kasuto H, Levavi B. Development of specific enzymelinked immune-osorbent assay for determining LH and FSH levels in tilapia, using recombinant gonadotropins. Gen Comp Endocrinol, 2007, 153(1 3): Ando H, Swanson P, Urano A. Regulation of LH synthesis and release by GnRH and gonadal steroids in masu salmon. Fish Physiol Biochem, 2003, 28(1 4): Gen K, Okuzawa K, Senthiekumaran B, et al. Unique expression of gonadotropin-i and-ii subunit genes in male and female red seabream (Pagrus major) during sexual maturation. Biol Reprod, 2000, 63(1): Gomez JM, Weil C, Ollitrault M. Growth hormone (GH) and gonadotropin subunit gene expression and pituitary and plasma changes during spermatogenesis and oogenesis in rainbow trout (Oncorhynchus mykiss). Gen Comp Endocrinol, 1999, 113(3): Halban PA, Irminger JC. Sorting and processing of secretory proteins. Biochem, 1994, 299(pt1): 1 18 Hassin S, Elizur A, Zohar Y. Molecular cloning and sequence analysis of striped bass (Morones axatilis) gonadotrophin-i and -II subunits. Mol Endocrinol, 1995, 15(1): Hellqvist A, Schmitz M, Mayer I, et al. Seasonal changes in expression of LH-β and FSH-β in male and female three-spined stickleback, Gasterosteus aculeatus. Gen Comp Endocrinol, 2006, 145(3): Hirohiko K, Koichiro G, Koichi O, et al. Effects of luteinizing hormone and follicle-stimulating hormone and insulin-like growth factor on aromatase activity and P450 aromatase gene expression in the ovarian follicles of red seabream, Pagrus major. Biol Reprod, 2003, 68(5): Jackson K, Goldberg D, Ofir M, et al. Blue gourami (Trichogaster trichopterus) gonadotropic β subunits (I and II) cdna sequences and expression during oogenesis. J Mol Endocrinol, 1999, 23: Kagawa H, TanakaH, Okuzawa K, et al. GTH II but not GTH I induces final maturation and the development of maturational competenceof oocytes of red seabreamin vitro. Gen Comp Endocrinol, 1998, 112(1): Kajimura S, Yoshiura Y, Suzuki M, et al. cdna cloning of two gonadotropin b subunits (GTH-I b and II b) and their expression profiles during gametogenesis in the Japanese flounder (Paralichthys olivaceus). Gen Comp Endocrinol, 2001, 122: Kobayashi M, Kato Y, Yoshiura Y, et al. Molecular cloning of the cdnas encoding two gonadotropin β subunits (GTH-I β and II β) from the goldfish, Carassius auratus. Gen Comp Endocrinol, 1997, 105(3): Lange IG, Hartel A, Meyer HHD. Evolution of oestrogen function in vertebrates. J Steroid Biochem, 2003, 83(1 5): Meiri I, Knibb WR, Zohar Y, et al. Temporal profile of folliclestimulating hormone, luteinizing hormone, and growth hormone gene expression in the protandrous hermaphrodite, gilthead seabream, Sparus aurata. Gen Comp Endocrinol, 2004, 137(3): Methven DA, Crim LW, Norberg B, et al. Seasonal reproduction and plasma levels of sex steroids and vitellogenin in Atlantic halibut (Hippoglossus hippoglossus). Can J Fish Aquat Sci, 1992, 49(4): Miranda LA, Strüssmann CA, Guilgur LG, et al. Cloning of FSH-b, LH-b and glycoprotein hormone subunits in pejerrey Odontesthes bonariensis (Valenciennes): expression profile and relationship with GnRH expression and plasma sex steroid levels in male fish. J Fish Biol, 2007, 71(6): Nyuji M, Shiraishi T, Selvaraj S, et al. Immunoreactive changes in pituitary FSH and LH cells during seasonal reproductive and spawning cycles of female chub mackerel Scomber japonicas. Fish Sci, 2011, 77(5): Picha ME, Shi B, Thomas P. Dual role of IGF-II in oocyte maturation in southern flounder Paralichthys lethostigma: Up-regulation of mpra and resumption of meiosis. Gen

10 4 : (Paralichthys lethostigma)lh β 31 Comp Endocrinol, 2012, 177(2): Pierce JG, Parsons TF. Glycoprotein hormones: structure and function. Annu Rev Biochem, 1981, 50: Sekine S, Saito A, Itoh H, et al. Molecular cloning and sequence analysis of chum salmon gonadotropin cdnas. Proc Natl- Acad Sci USA, 1989, 86(22): Shimon H, Claire HH, Yonathan Z. Ontogeny of follicle-stimulating hormone and luteinizing hormone gene expression during pubertal development in the female striped bass, Morone saxatilis (Teleostei). Biol Reprod, 1999, 61(6): So WK, Kwok HF, Ge W. Zebrafish gonadotropins and their Receptors: II. Cloning and characterization of zebrafish folliclestimulating hormone and luteinizing hormone subunits their spatial-temporal expression patterns and receptor specificity. Biology of Reproduction, 2005, 72(6): Sohn YC, Suetake H, Yoshiura Y, et al. Structural and expression analyses of gonadotropin I beta subunit genes in goldfish (Carassius auratus). Gene, 1998, 222(2): Swanson P, Dickey JT, Campbell B. Biochemistry and physiology of fish gonadotropins. Fish Physiol Biochem, 2003, 28(1 4): Trinh KY, Wang NC, Hew CL, et al. Molecular cloning and sequencing of salmon gonadotropin subunit. Eur J Biochem, 1986, 159(3): Tubbs C, Tan WX, Shi B, et al. Identification of 17, 20 beta, 21- trihydroxy-4-pregnen-3-one (20 beta-s) receptor binding and membraneprogestin receptor alpha on southern flounder sperm (Paralichthys lethostigma) and their likely role in 20 beta-s stimulation of sperm hypermotility. General and Comparative Endocrinology, 2011, 170(3): Weil C, Bougoussa HM, Gallais C, et al. Preliminary evidence suggesting variations of GtH 1 and GtH 2 mrna levels at different stages of gonadal development in rainbow trout, Oncorhyn chusmykiss. Gen Comp Endocrinol, 1995, 100(3): Yoshiura Y, SuetakeH, Aida K. Duality of gonadotropin in a primitive teleost, Japanese eel(anguilla japonica). Mol Endocrinol, 1996, 16(2): ( ) Molecular Cloning and Differential Expression of LH β Subunit During the Ovarian Maturation Cycle of Paralichthys lethostigma LIU Xuezhou 1, ZANG Kun 1, 2, XU Yongjiang 1, LI Chunguang 1, SHI Bao 1, WANG Yanyan 1 (1. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao ; 2. College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai ) Abstract The role of the endocrine pathway in the regulation of southern flounder Paralichthys lethostigma reproduction has been unclear. Here we aimed to explore the underlying mechanisms by investigating the expression pattern of luteinizing hormone β subunit (LH β). Using samples isolated from pituitary of P. we obtained the full-length cdna sequence of LH β with the RACE method. The spatial and temporal expression patterns of LH β mrna in different tissues during an ovarian maturation cycle were analyzed using the qpcr assay. The levels of plasma LH and E 2 during an ovarian maturation cycle were also measured using the ELISA method. Our results revealed an asynchronous oocyte development and batch spawning in southern flounder. The cdna sequence of LH β had a length of 597 bp and it encoded 145 amino acids. LH β expression was detected in all examined tissues except for muscle, which implied it could be involved in multiple physiological functions in southern flounder. The LH β mrna level in pituitary was high through the entire ovarian maturation cycle, and peaked at stage V and VI. In the ovary the LH β mrna level peaked at stage V. In the liver the LH β mrna level was the highest at stage II and relatively high at stage V. The levels of plasma LH and E 2 fluctuated in a similar pattern during the ovarian maturation cycle. These results suggested that LH β might regulate the ovarian maturation cycle via endocrine, paracrine and autocrine pathways. Our study provided insights into the functions of the endocrine pathway in the reproduction of southern flounder. Key words Paralichthys lethostigma; LH β subunit; Ovarian maturation cycle; Expression pattern; Plasma hormone levels

Microsoft Word - 20 徐永江.doc

Microsoft Word - 20 徐永江.doc 43 6 Vol.43, No.6 2012 11 OCEANOLOGIA ET LIMNOLOGIA SINICA Nov., 2012 (Verasper moseri) * 1, 2 1, 2 1, 2 1, 2 (1. 266071; 2. 266071) (sgnrha) (HCG), : sgnrha HCG,, (T) (E 2 ), 96h, sgnrha, HCG,,,, S965