48 (6) : , 2002 L H, (Chapple, 1991 ; Shoham, 1993) ( Howles,, 1986 ; Conway, 1989 ; Regan, 1989) ;, (Westergarrd, 1999)

Transcription

1 48 (6) : , 2002 A cta Zoologica S inica 3 (, ) (r2hl H), r2hfsh (, 20 IU) r2hl H (0 20 IU), r2hl H (5 20 IU), r2 hl H (15 IU), ;,, PMSG2hCG,, r2hl H r2hl H 15 IU, PMSG2hCG, r2hl H ( ), r2hl H ( FSH) (L H),,, L H, (r2 (Chapple, 1991 ; Shoham, 1993) :, L H, ( Howles,, hl H 1986 ; Conway, 1989 ; Regan, 1989) ;, L H, (Westergarrd, 1999),,, 111 ( ) ( Sawitzke, 1991), (20, g), ( ), : ( PMSG, ( Sawitzke, 1991 ; Shoham, 1996), ), ( r2hfsh,, Serono, ) ( r2hl H, (r2hfsh r2hl H), L H, 112 (Balasch, 1995 ; Anderson, 1999) DNA hfsh) (r2hl H), 1 ), ( hcg, Serono, ) PMSG2hCG, PMSG 43 h, 100 %,, hcg, 5 h r2hfsh r2hl H , (No ) 33 E2mail : 37,, :

2 6 : 755 ( ), ( P < 0101, 2) r2hl H, Zeibe Andersen ( Flemming, 1998), r2hfsh r2hfsh (20 IU) r2hl H (5 20 IU) 43 h, r2hl H, 5 h 113 Anderson et al. (1999),, Petri dish ( Nunc, ), l (40 50 ) ; EBSS ( Hyclone, ) g/ L ( ) + 11 mg/ L ( Gibcol BRL, ) % ( Gibcol BRL, ) + ( ) ; IVF Medium ( MEDI2CUL T Universal, ) + 114, IVF Medium, 2 r2hfsh r2hl H 37, 5 % CO 2 95 % CO 2, 48 h, h Table 2 Effect of IVC on early embryos in immature mice, superovulated with a dose of r2hfsh and r2hl H, Sigma Plot t P < , r2hfsh r2hl H r2hfsh ( 20 IU) r2hl H (5 20 IU), 5 IU r2hl H 20 IU r2 hl H , PMSG2hCG ( ), ( P < 0101) ; 10 IU 15 IU r2 1 r2hfsh r2hl H Table 1 Effect of superovulation on immature mice Dosage of hormone with a dose of r2hfsh and r2hl H ( ) ( ) hl H, ( P > 0105) r2 hfsh (20 IU) r2hl H (20 IU) r2hfsh r2hl H r2hl H, ( 1), PMSG2hCG, ( P > 0105, 3) h, ( PMSG2hCG) r2hfsh r2hl H ( ), h, % %, r2hl H PMSG2hCG 4 5 IU r2hl H 15 IU 20 IU r2hl H % 0 % ; r2hl H 15 IU,, 3519 % 7196 %,, 8111 % 2134 % % ( P < 0101) Number of mice Number of preembryos/ mouse F + 0 IU r2hl H 15 0 a F + 20 IU r2hl H 15 0 a F + 5 IU r2hl H b F + 10 IU r2hl H c F + 15 IU r2hl H c F + 20 IU r2hl H d 5 IU PMSG + 5 IU hcg c Morulae and blastocyst rate Blastocyst rate F + 5 IU r2l H a (5/ 47) 0 (0/ 47) a F + 10 IU r2l H b (24/ 105) 0 (0/ 105) a F + 15 IU r2l H b (42/ 117) 7196 (9/ 117) b F + 20 IU r2l H b (34/ 155) 1194 (3/ 155) a 5 IU PMSG + 5 IU hcg c (69/ 116) (37/ 116) c 212 r2hfsh r2hl H r2hl

3 r2hfsh r2hl H IU r2hl H, FSH 012 IU hcg, Table 3 Effect of superovulation on early, immature mice with a dose of r2hfsh, 20 IU r2hfsh + combining with a ratio of r2hl H 5 IU r2hl H, ( ) r2hl H ( Number of mice ( ) Number of preembryos/ mouse ( F + 5 IU r2hl H) + 15 IU r2hl H a ( F + 10 IU r2hl H) + 15 IU r2hl H a ( F + 15 IU r2hl H) + 15 IU r2hl H a ( F + 20 IU r2hl H) + 15 IU r2hl H a 5 IU PMSG + 5 IU hcg a (Same superscript indicates no significant difference) ( P > 0105) 4 r2hfsh r2hl Hr2hFSH, Table 4 Effect of IVC on early embryos in immature mice ( 3), r2hfsh superovulated with a dose of r2hfsh combining with a ratio of r2hl H Morulae and blastocyst rate Blastocyst rate ( F + 5 IU r2hl H) + 15 IU r2hl H a (18/ 101) 0 (0/ 101) a ( F + 10 IU r2hl H) + 15 IU r2hl H b (26/ 111) 0 (0/ 111) a ( F + 15 IU r2hl H) + 15 IU r2hl H b (28/ 111) 8111 b (9/ 111) ( F + 20 IU r2hl H) + 15 IU r2hl H a (21/ 128) 2134 a (3/ 128) 5 IU PMSG + 5 IU hcg c (69/ 116) c (37/ 116) 3 ( ), 311, Anderson et al. (1999) Flemmning et al. (1998) 20 IU r2hfsh 5 20 IU r2hl H,, L H, r2hl H,, ( 1) r2hl H 10 IU, PMSG2hCG, r2hl H 20 IU, FSH, ( , P < 0101) Anderson et al1 (1999) FSH (20 IU) 20 1), r2hl H,,, ( Westergaard, 1999) Schramn et al. (1994), ( IVM), hfsh (110 m g/ ml) hl H (110 m g/ ml),,, hfsh hl H IVM r2hl H, ( < 20 IU), r2hl H,, Schramn 312 r2hfsh r2hl H,, h,, r2hl H ( 3 4), 15 IU r2hl H,,, r2hl H, L H, L H (, 2001), PMSG2hC G

4 6 : 757,, r2hl H,, hcg hl H, Anderson et al. (1999) r2 ( r2hfsh r2 hl H hcg, hl H) : r2hl H (, Jamson et ), r2hl H al. (1986) Talmade et al. (1984), hl H hcg, hcg hl H, ( References) Anderson, C. Y., S. Ziebe and G. L. Xia 1999 Requirements for human chorionic gonadotropin and recombinant human luteinizing hormone for follicular development and maturation. J. Assit. Reprod. Genet. 16 : Balasch, J., F. Miro and I. Burzaco 1995 The role of luteinizing hormone in human follicle development and oocyte fertility : evidence from i n2 vit ro fertilization in a woman with long2standing hypogonadotrophic hypogonadism and using recombinant human follicle stimulating hormone. Hum. Reprod. 10 : Chapple, S. and C. Howles 1991 Reevaluation of the roles of luteinizing hormone and follicle2stimulating hormone in the ovulatory process. Hum. Reprod. 6 : Chen, Y., G. L. Xia, Y. Q. Su, H. B. Wang and C. Hao 2001 Effect of gonadotropinson i n vit ro maturation of oocyte and cumulus ex2 pansion in kunming white mouse. Acta Zool. Si n. 47 (2) : [,,,, (2) : ] Conway, G. S., J. W. Honour and H. S. Jacobs 1989 Heterogeneity of the polycystic ovary syndrome : clinical, endocrine and ultrasound features. Cli n. Endocri nol. 30 : Flemming, R., F. Lloyd and M. Herbert 1998 Effects of profound suppression on luteinizing hormone during ovarian stimulation on follicular activity, oocyte and embryo function in cycles stimulated with purified follicular stimulating hormone. Hum. Reprod. 13 : Howles, C., M. C. Macname and R. G. Edwards 1986 Effect of high tonic levels of luteinizing hormone on outcome of i n vit ro fertilization. L ancet 2 : Jameson, J. L. and J. F. Lindell 1986 Evolution of different transcriptional start sites in the human luteinizing hormone and chorionic go2 nadotropin 2subunit genes. DNA 5 : Regan, L., E. J. Owen and H. S. Jacobs 1989 Hypersecretion of luteinizing hormone, infertility and miscarriage. L ancet 336 : Sawitzke, A. L. and J. Griffin 1991 Purified preparation of hl H are contaminated with small amount of a chorionic gonadotropin2like material. J. Cli n. Endocri nol. Metab. 72 : Shoham, Z., H. S. Jacobs and V. Insler 1993 Luteinizing hormone : its role, mechanism of action, and detrimental effects when hypersecret2 ed during the follicular phase. Fertil. Steril. 59 : Shoham, Z. and V Insler 1996 Recombinant technique and gonadotropins production : era in reproductive medicine. Fertil. Steril. 66 : 193 Westergaard, L. G Clinical outcome of i n2vit ro fertilization : hm G versus purified FSH. In : Fillicori, M. ed. The Role of Luteinizing Hormone in Folliculogenesis and Ovulation. Bologna, Italy : Monduzzi Editori,

5 ( Abstract) INFL UENCE OF RECOMBINANT HUMAN L UTEINIZING HORMONE ON THE DEVELOPMENT OF EARLY EMBRYOS IN MICE 3 GUO Yong XIA Guo2Liang 33 WAN G Hai2Bin XIE Hui2Rong L EI Lei BO Shu2Min ( College of Biological Science, Chi na A gricult ural U niversity, Beiji ng , Chi na) Although the biochemical purified gonadotropins have been utilized mainly in the studies on the mammalsπ reproductive processes (including t he early embryonic develop ment), t he problem of t hose so2called contami2 nants in gonadotropins purified by such a way has still existed. It means that these contaminants might affect the real function of those biochemically purified hormones on the mammalsπ propagation. On the other hand, by t he recombinant DNA technology, t heir recombinant gonadot ropinic product s could escape most of t he prob2 lems in t he regularly biochemical purified ones. sary for t he effective research work on mammalsπ reproductive processes. The biological stability of such recombinant hormones is neces2 In t his experiment, bot h t he recombi2 nant human follicular stimulating hormone ( r2hfsh) and recombinant human luteinizing hormone ( r2hl H) were applied for superovulating t he immat ure prepubertal Kunming mice, t hen, t hose flushed preembryos were i n vit ro cult ured for observation of t heir blastocyst formation capacity. In groups superovulating wit h r2hfsh (20 IU) and r2hl H (0 20 IU), the number of preembryos for mice was in dose2dependent with r2hl H ( to ), their blastocystal rates were also in such a relation with it (0 % 7196 %). In groups su2 perovulating with r2hfsh (20 IU) combining a ratio of r2hl H (0 20 IU), the number of preembryos for mice was similar among t hose different groups, but t heir blastocyst rates were somewhat dose2dependent wit h r2 hl H ( %). The results showed that r2hl H probably affect the early embryonic development in mice negatively, especially for t heir blastocystal formation. Key words Mice, Early embryonic development, Recombinant human follicular stimulating hormone, Re2 combinant human luteinizing hormone 3 This work was supported by a grant from National Natural Science Foundation of China (No ) 33 Corresponding author. cau. edu. cn