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Biochemical Compositions of Follicular Fluid and the Effects of Culture Conditions on the In Vitro Development of Pig Oocytes
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Biochemical Compositions of Follicular Fluid and the Effects of Culture Conditions on the In Vitro Development of Pig Oocytes
Huang, Wei-Tung; Lu, She-Ghi; Tang, Pin-Chi; Wu, Shinn-Chih; Cheng, San-Pao; Ju, Jyh-Cherng;
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The aims of this study were, firstly, to analyze the biochemical compositions of serum and follicular fluid (FF) from prepubertal gilts after PMSG (1,000 IU) treatment. The concentrations of total proteins, lipids, cholesterol, glucose and sex hormones (progesterone, ; estradiol-, ; testosterone, T) were measured. Secondary, the effects of porcine FF (pFF) addition (40% and 100%) in IVM media and different culture conditions [Exp. 1: mBMOC-2+20% porcine serum (PS), fresh IVM medium, filtered IVMconditioned medium, or rabbit oviducts; Exp. 2: mBMOC-2+20%PS or stepwise medium replacement procedures (SMRP) cocultured with or without cumulus cells] on the in vitro development (IVD) of porcine oocytes were also examined. Results showed that no significant differences were found in total protein levels between serum and pFF from different sizes (large, >7 mm; medium, ~5-7 mm; small, <3-5 mm) of follicles (75-85 and 49-90 mg/dl; p>0.05). Total lipid concentrations remained constant in serum (395-472 mg/dl), and reduced significantly in the pFF from large follicles (287 mg/dl) at 132 h after PMSG treatment when compared to those at other time points (441-480 mg/dl). Basal cholesterol levels in serum and pFF at 12 h were similar (153-161 mg/dl), but increased at 36 h (186-197 mg/dl). Basal P4 and E2 levels in serum (0.1 ng/ml and 5.5 pg/ml) were low, but increased from 0.34 ng/ml and 12.13 pg/ml at 24 h to 0.81 ng/ml and 61.70 pg/ml at 98 h, respectively, after PMSG treatment (p<0.05). P4 levels increased linearly in pFF from large follicles during 12 through 132 h (138-1,288 ng/ml). A similar increase was also observed in levels (22-730 pg/ml) before 60 h post PMSG treatment, and then dropped afterwards (730-121 pg/ml). The development of the oocytes fertilized in 40% pFF-medium was greater than that in 100% pFF-medium group without gonaodtropin addition (31% vs 10%, p<0.05). However, both were lower than those in mBMOC-2+20%PS and in rabbit oviducts (p<0.05). When cocultured with cumulus cell monolayers, a greater cleavage rate was observed in the group cultured in filtered IVM-conditioned medium than the SMRP group (36% vs 18%, p<0.05). A similar phenomenon was also observed in the culture without cumulus cell monolayers (33% vs 19%, p<0.05). It is concluded that neither the fresh IVM nor filtered IVM-conditioned medium has positive effect on the IVD of oocytes. Coculture with cumulus cell monolayers and the SMRP were not beneficial to the development of IVF pig oocytes.
Protein;Lipid;Cholesterol;Sex Hormones;IVMFC;
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Ainsworth, L., B. K. Tsang, B. R. Downey, G. J. Marcus and D. T. Armstrong. 1980. Interrelationships between follicular fluid steroid levels, gonadotropic stimuli, and oocyte maturation during preovulatory development of porcine follicle. Biol. Reprod. 23:621-627. crossref(new window)

Agrawal, K. P., I. V. Mongha and N. K. Bhattacharyya. 1983. Survival of goat embryos in rabbit oviduct. Vet. Rec. 112:200. crossref(new window)

Allen, W. R., F. Stewart, A. O. Trounson, M. Tischner and W. Bielanski. 1976. Viability of horse embryos after long distance transport in the rabbit. J. Reprod. Fertil. 47:387-390. crossref(new window)

Archibong, A. E., R. M. Petters and B. H. Johnson. 1989. Development of porcine embryos from one- and two-cell stages to blastocysts in culture medium supplemented with porcine oviductal fluid. Biol. Reprod. 41:1076-1083. crossref(new window)

Barboni, B., M. Turriani, G. Galeati, M. Spinaci, M. L. Bacci, M. Forni and M. Mattioli. 2000. Vascular endothelial growth factor production in growing pig antral follicles. Biol. Reprod. 62:1160-1167. crossref(new window)

Brussow, K. P., J. Ratkyand, F. Schneider, H. Torner, W. Kanitz and L. Solti. 1999. Effects of follicular fluid on the transport of porcine oocytes into the oviduct at ovulation. Reprod. Dom. Anim. 34:423-429. crossref(new window)

Burr, J. H. and J. I. Davies. 1951. The vascular system of the rabbit ovary and its relationship to ovulation. Anat. Rec. 111:273-297. crossref(new window)

Coy, P., S. Ruiz, R. Romar, I. Campos and J. Gadea. 1999. Maturation, fertilization and complete development of porcine oocytes matured under different systems. Theriogenology 514:799-812. crossref(new window)

Davis, D. L. and B. N. Day. 1978. Cleavage and blastocyst formation by pig eggs in vitro. J. Anim. Sci. 46:1043-1053. crossref(new window)

Ding, J. and G. R. Foxcroft. 1992. Follicular heterogeneity and oocyte maturation in vitro in pigs. Biol. Reprod. 47:648-655. crossref(new window)

Ellefson, R. D. and W. T. Caraway. 1975. Lipids and lipoproteins. In: 'Foundamentals of Chemical Chemistry' (Ed. N. W. Tiete and W. B. Sauders). Co. Philadelphia.

Espey, L. L. and H. Lipner. 1994. Ovulation. In: 'The Physiology of Reproduction'. 2nd edition (Ed. E. Knobil and J. D. Neill). Raven Press. pp. 725-780.

Falhot, K., B. Lund and W. Falhot. 1973. An easy colorimetric micromethod for routine determination of free fatty acid in plasma. Clinica Chimica Act. 46:105. crossref(new window)

Fukushima, M. and Y. Fukui. 1985. Effects of gonadotropins and steroids on the subsequent fertilizability of extra-follicular bovine oocytes cultured in vitro. Anim. Reprod. Sci. 9:323-332. crossref(new window)

Funahashi, H. and B. N. Day. 1993a. Effects of follicular fluid at fertilization in vitro on sperm penetration in pig oocytes. J. Reprod. Fertil. 99:97-103. crossref(new window)

Funahashi, H. and B. N. Day. 1993b. Effects of different serum supplements in maturation medium on meiotic and cytoplasmic maturation of pig oocytes. Theriogenology 39:965-973. crossref(new window)

Gerard, M., Y. Menezo, P. Rombauts, D. Szollosi and C. Thibault. 1979. In vitro studies of oocytes of oocyte maturation and follicular metabolism in the pig. Annls. Biol. Anim. Biochim. Biophys. 19:1521-1535. crossref(new window)

Guraya, S. S. 1985. Biology of ovarian follicles in mammals. Springer-Verlag, Berlin. pp. 150-194.

Hafez, E. S. E. 1987. Folliculogenesis, egg maturation and ovulation. In: Reproduction in Farm Animals (Ed. E. S. E. Hafez). 5th edtion. Lea & Febiger, Philadelphia. pp.130-167.

Herrmann, H. H. and W. Holtz. 1985. Storage of pig embryos in the ligated rabbit oviduct and its effect on the viability after retransfer to synchronized gilts. Anim. Reprod. Sci. 8:159-170. crossref(new window)

Huang, W. T., P. C. Tang, S. C. Wu, S. P. Cheng and J. C. Ju. 2001. Effects of levels and sources of follicular fluid on the in vitro maturation and development of porcine oocytes. Asian-Aust. J. Anim. Sci. 14(10):1360-1366. crossref(new window)

Hunter, M. G. 2000. Oocyte maturation and ovum quality in pigs. Rev. Reprod. 5:122-130. crossref(new window)

Hunter, R. H. F. 1990. Fertilization of pig eggs in vivo and in vitro. J. Reprod. Fertil. Suppl. 40:211-226.

Ju, J. C., Y. C. Chang, W. T. Huang, P. C. Tang and S. P. Cheng. 2001. Superovulation and transplantation of demi- and aggregated-embryos in rabbits. Asian-Aust. J. Anim. Sci. 14:455-461. crossref(new window)

Ju, J. C., S. P. Cheng, P. C. Tarng and K. B. Choo. 1991. In vivo development and microinjection of rabbit zygotes. Asian-Aust. J. Anim. Sci. 4(1):73-78. crossref(new window)

Kihara, T., A. Kimura, A. Moriyama, I. Ohkubo and T. Takahashi. 2000. Identification of components of the intrafollicular bradykinin-producing system in the porcine ovary. Biol. Reprod. 63:858-864. crossref(new window)

Kishi, H., M. Kondoh, N. Nagamine, F. Shi, G. Watanabe and K. Taya. 1997. Roles of the basal level of LH and FSH in the regulation of follicular development during pseudopregnancy in the rat. J. Reprod. Dev. 43(4):279-287. crossref(new window)

Krisher, R. L., R. M. Petters and B. H. Johnson. 1989. Effect of oviductal condition on the development of one-cell porcine embryos in mouse or rat oviducts maintained in organ culture. Theriogenology 32:885-892. crossref(new window)

Lin, J. H., T. Shiao and L. A. Shie. 1977. Ultra microtechnique for determination of blood glucose with the modified Somogyi-Nelson method. J. Chin. Soc. Anim. Sci. 6:57-59.

Linder, G. M. and R. W. Wright, Jr. 1978. Morphological and quantitative aspects of the development of swine embryo in vitro. J. Anim. Sci. 46:711-718. crossref(new window)

Lipner, H. and R. O. Greep. 1971. Inhibition of steroidogenesis at various sites in the biosynthetic pathway in relation to induced ovulation. Endocrinology 88602-604. crossref(new window)

Long, C. R., J. R. Dobrinsky and L. A. Johnson. 1999. In vitro production of pig embryos: comparisons of culture media and boars. Theriogenology 51:1375-1390 crossref(new window)

Lowry, O. H., N. J. Rosebrough, A. L. Farr and R. J. Randall. 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193:265-267.

Machaty, Z., B. N. Day and R. S. Prather. 1998. Development of early porcine embryos in vitro and in vivo. Biol. Reprod. 59:451-455. crossref(new window)

Meinecke, B. and S. Meinecke-Tillmann. 1998. Amino acid concentrations in porcine follicular fluid and ovarian plasma during preovulatory follicular development. Tieraerztl. Umschau. 53(7):429-436.

Nagai T, K. Niwa and A. Iritani. 1984. Effect of sperm concentration during preincubation in a defined medium on fertilization in vitro of pig follicular oocytes. J. Reprod. Fertil. 70:271-275. crossref(new window)

Naito, K., Y. Fukuda and Y. Toyoda. 1988. Effects of porcine follicular fluid on male pronucleus formation in porcine oocytes matured in vitro. Gamete Res. 21:289-295. crossref(new window)

Naito, K., Y. Fukuda and I. Ishibashi. 1989. Developmental ability of porcine ova matured in porcine follicular fluid in vitro and fertilized in vitro. Theriogenology 31:1049-1057. crossref(new window)

Naito, K., M. Kosaka, Y. Fukuda, I. Ishibashi and Y. Toyoda. 1990. Analysis of the factor(s) present in follicular fluids promoting male pronucleus formation ability of porcine follicular oocytes. Jpn. J. Anim. Reprod. 36:213-218. crossref(new window)

Petters, R. M. and K. D. Wells. 1993. Culture of pig embryos. J. Reprod. Fertil. Suppl. 48:61-73.

Petters, R. M., B. H. Johnson, M. L. Reed and A. E. Archibong. 1990. Glucose, glutamine and inorganic phosphate in early development of the pig embryo in vitro. J. Reprod. Fertil. 89:269-275. crossref(new window)

Pope, C. E. and B. N. Day. 1977. Transfer of preimplantation pig embryos following in vitro culture for 24 or 48 hours. J. Anim. Sci. 44:1036-1040. crossref(new window)

Reed, M. L., M. J. Illera and R. M. Petters. 1992. In vitro culture of pig embryos. Theriogenology 37:95-109. crossref(new window)

Rondell, P. 1970. Follicular process in ovulation. Fed. Pro. 29:1875.

Sirard, M. A. and R. D. Lambert. 1986. Birth of calves after in vitro fertilization using laparoscopy and rabbit oviduct incubation of zygotes. Vet. Rec. 119:167-169. crossref(new window)

Snedecodr, G. W. and W. G. Cochran. 1980. Statistical methods.7th edition. Ames IA, The Iowa State Univ. Press, pp. 290-291.

Tsafriri, A. and C. P. Channing. 1975. An inhibitory influence of granulosa cells and follicular fluid upon porcine oocyte meiosis in vitro. Endocrinology 96:922-927. crossref(new window)

Vatzias, G. and D. R. Hagen. 1999. Effects of porcine follicular fluid and oviduct-conditioned media on maturation and fertilization of porcine oocytes in vitro. Biol. Reprod. 60:42-48. crossref(new window)

White, K. L., K. Hehnke, L. F. Rickords, L. L. Southern, D. L. Thompson, Jr. and T. C. Wood. 1989. Early embryonic development in vitro by coculture with oviductal epithelial cells in pigs. Biol. Reprod. 41:425-430. crossref(new window)

Wright, Jr. R. J. 1977. Successful culture in vitro of swine embryo to the blastocyst stage. J. Anim. Sci. 44:854-858. crossref(new window)

Wu, S. C., S. P. Cheng, B. T. Liu, C. M. Chiou and C. C. Chu. 1991. In vitro maturation of porcine oocytes and their subsequent developmental capacity: II. Effects of the follicle size and pFF addition to the oocyte maturation medium. J. Chinese Soc. Anim. Sci. 20:469-480.

Yamada, Y. and Y. Kawai. 1997. Changes of sex steroids and prostaglandins around induced ovulation. Jpn. J. Swine. Sci. 34(4):157-162. crossref(new window)

Yamada, Y. and Y. Kawai. 1997. Changes of sex steroids and prostaglandins at inhibition of ovulation by inhibitor of prostaglandin biosynthesis in the gilts. Jpn. J. Swine. Sci. 36(3):117-123.

Yanagimachi, R. 1969. In vitro capacitation of hamster spermatozoa by follicular fluid. J. Reprod. Fertil. 18:275-286. crossref(new window)

Yoshida, M., Y. Ishizaki and H. Kawagishi. 1990. Blastocyst formation by pig embryos resulting from in-vitro fertilization of oocytes matured in vitro. J. Reprod. Fertil. 88:1-8. crossref(new window)

Yoshida, M., Y. Ishizaki, H. Kawagishi, K. Bamba and Y. Kojima. 1992. Effects of pig follicular fluid on maturation of pig oocytes in vitro and on their subsequent fertilizing and developmental capacity in vitro. J. Reprod. Fertil. 95:481-488. crossref(new window)