Abstract
Selecting oocytes that are most likely to develop is crucial for in vitro fertilization and animal cloning. Brilliant cresyl blue (BCB) staining has been used for oocyte selection in large animals, but its wider utility needs further evaluation. Mouse oocytes were divided into those stained (BCB+) and those unstained (BCB−) according to their ooplasm BCB coloration. Chromatin configurations, cumulus cell apoptosis, cytoplasmic maturity and developmental competence were compared between the BCB+ and BCB− oocytes. The effects of oocyte diameter, sexual maturity and gonadotropin stimulation on the competence of BCB+ oocytes were also analyzed. In the large- and medium-size groups, BCB+ oocytes were larger and showed more surrounded nucleoli (SN) chromatin configurations and higher frequencies of early atresia, and they also gained better cytoplasmic maturity (determined as the intracellular GSH level and pattern of mitochondrial distribution) and higher developmental potential after in vitro maturation (IVM) than the BCB− oocytes. Adult mice produced more BCB+ oocytes with higher competence than the prepubertal mice when not primed with PMSG. PMSG priming increased both proportion and developmental potency of BCB+ oocytes. The BCB+ oocytes in the large-size group showed more SN chromatin configurations, better cytoplasmic maturity and higher developmental potential than their counterparts in the medium-size group. It is concluded that BCB staining can be used as an efficient method for oocyte selection, but that the competence of the BCB+ oocytes may vary with oocyte diameter, animal sexual maturity and gonadotropin stimulation. Taken together, the series of criteria described here would allow for better choices in selecting oocytes for better development.
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References
Sirard MA, Blondin P . Oocyte maturation and IVF in cattle. Anim Reprod Sci 1996; 42:417–426.
Cha KY, Han SY, Chung HM, et al. Pregnancies and deliveries after in vitro maturation culture followed by in vitro fertilization and embryo transfer without stimulation in women with polycystic ovary syndrome. Fertil Steril 2000; 73:978–983.
Zhang X, Zerafa A, Wong J, Armstrong DT, Khamsi F . Human menopausal gonadotropin during in vitro maturation of human oocytes retrieved from small follicles enhances in vitro fertilization and cleavage rates. Fertil Steril 1993; 59:850–853.
Trounson A, Wood C, Kausche A . In vitro maturation and the fertilization and developmental competence of oocytes recovered from untreated polycystic ovarian patients. Fertil Steril 1994; 62:353–362.
Trounson A, Anderiesz C, Jones GM, Kausche A, Lolatgis N, Wood C . Oocyte maturation. Hum Reprod 1998; 13(Suppl 3):52–62.
Barnes FL, Crombie A, Gardner DK, et al. Blastocyst development and birth after in-vitro maturation of human primary oocytes, intracytoplasmic sperm injection and assisted hatching. Hum Reprod 1995; 10:3243–3247.
Barnes FL, Kausche A, Tiglias J, Wood C, Wilton L, Trounson A . Production of embryos from in vitro-matured primary human oocytes. Fertil Steril 1996; 65:1151–1156.
Tan JH, Yang ZM, Qin PC, Pashen R . Light and electron microscope studies on follicular oocytes of Chinese yellow cattle prior to in vitro maturation (Abst.) Theriogenology 1988; 29:317.
Gordon I . Laboratory Production of Cattle Embryos (Biotechnology in Agriculture No. 27). 2nd Edition, Cambridge, UK: CAB International/Cambridge University Press, 2003.
Pavlok A, Lucas-Hahn A, Niemann H . Fertilization and developmental competence of bovine oocytes derived from different categories of antral follicles. Mol Reprod Dev 1992; 31:63–67.
Hendriksen PJ, Vos PL, Steenweg WN, Bevers MM, Dieleman SJ . Bovine follicular development and its effect on the in vitro competence of oocytes. Theriogenology 2000; 53:11–20.
Hyttel P, Fair T, Callesen H, Greve T . Oocyte growth, capacitation and final maturation in cattle. Theriogenology 1997; 47:23–32.
Ma SF, Lan GC, Miao YL, et al. Hypoxanthine (HX) inhibition of in vitro meiotic resumption in goat oocytes. Mol Reprod Dev 2003; 66:306–313.
Revel F, Mermillod P, Peynot N, Renard JP, Heyman Y . Low developmental capacity of in vitro matured and fertilized oocytes from calves compared with that of cows. J Reprod Fertil 1995; 103:115–120.
Damiani P, Fissore RA, Cibelli JB, et al. Evaluation of developmental competence, nuclear and ooplasmic maturation of calf oocytes. Mol Reprod Dev 1996; 45:521–534.
Khatir H, Lonergan P, Carolan C, Mermillod P . Prepubertal bovine oocyte: a negative model for studying oocyte developmental competence. Mol Reprod Dev 1996; 45:231–239.
Pujol M, Lopez-Bejar M, Paramio MT . Developmental competence of heifer oocytes selected using the brilliant cresyl blue (BCB) test. Theriogenology 2004; 61:735–744.
O'Brien JK, Dwarte D, Ryan JP, Maxwell WM, Evans G . Developmental capacity, energy metabolism and ultrastructure of mature oocytes from prepubertal and adult sheep. Reprod Fertil Dev 1996; 8:1029–1037.
Ledda S, Bogliolo L, Calvia P, Leoni G, Naitana S . Meiotic progression and developmental competence of oocytes collected from juvenile and adult ewes. J Reprod Fertil 1997; 109:73–78.
Pinkert CA, Kooyman DL, Baumgartner A, Keisler DH . In-vitro development of zygotes from superovulated prepubertal and mature gilts. J Reprod Fertil 1989; 87:63–66.
Izquierdo D, Villamediana P, Paramio MT . Effect of culture media on embryo development from prepubertal goat IVM-IVF oocytes. Theriogenology 1999; 52:847–861.
Koeman J, Keefer CL, Baldassarre H, Downey BR . Developmental competence of prepubertal and adult goat oocytes cultured in semi-defined media following laparoscopic recovery. Theriogenology 2003; 60:879–889.
Hagemann LJ, Beaumont SE, Berg M, et al. Development during single IVP of bovine oocytes from dissected follicles: interactive effects of estrous cycle stage, follicle size and atresia. Mol Reprod Dev 1999; 53:451–458.
Krisher RL . The effect of oocyte quality on development. J Anim Sci 2004; 82(E-Suppl): E14–E23.
Tsuji K, Sowa M, Nakano R . Relationship between human oocyte maturation and different follicular sizes. Biol Reprod 1985; 32:413–417.
Trounson A, Anderiesz C, Jones G . Maturation of human oocytes in vitro and their developmental competence. Reproduction 2001; 121:51–75.
Durinzi KL, Saniga EM, Lanzendorf SE . The relationship between size and maturation in vitro in the unstimulated human oocyte. Fertil Steril 1995; 63:404–406.
Russell JB . Immature oocyte retrieval combined with in-vitro oocyte maturation. Hum Reprod 1998; 13(Suppl 3):63–70.
Whitacre KS, Seifer DB, Friedman CI, et al. Effects of ovarian source, patient age, and menstrual cycle phase on in vitro maturation of immature human oocytes. Fertil Steril 1998; 70:1015–1021.
Debey P, Szollosi MS, Szollosi D, Vautier D, Girousse A, Besombes D . Competent mouse oocytes isolated from antral follicles exhibit different chromatin organization and follow different maturation dynamics. Mol Reprod Dev 1993; 36:59–74.
Zuccotti M, Piccinelli A, Rossi PG, Garagna S, Redi CA . Chromatin organization during mouse oocyte growth. Mol Reprod Dev 1995; 41:479–485.
Wassarman M . The mammalian ovum. In: Knobil E, Neil D, eds. The Physiology of Reproduction. Vol. 1, New York, USA: Raven Press, 1988:69–102.
Ericsson SA, Boice ML, Funahashi H, Day BN . Assessments of porcine oocytes using brilliant cresyl blue. Theriogenology 1993; 39:214 (abstr.).
Roca J, Martinez E, Vazquez JM, Lucas X . Selection of immature pig oocytes for homologous in vitro penetration assays with the brilliant cresyl blue test. Reprod Fertil Dev 1998; 10:479–485.
Rodriguez-Gonzalez E, Lopez-Bejar M, Velilla E, Paramio MT . Selection of prepubertal goat oocytes using the brilliant cresyl blue test. Theriogenology 2002; 57:1397–1409.
Alm H, Torner H, Lohrke B, Viergutz T, Ghoneim IM, Kanitz W . Bovine blastocyst development rate in vitro is influenced by selection of oocytes by brillant cresyl blue staining before IVM as indicator for glucose-6-phosphate dehydrogenase activity. Theriogenology 2005; 63:2194–2205.
De Schepper GG, Van Noorden CJ, Koperdraad F . A cytochemical method for measuring enzyme activity in individual preovulatory mouse oocytes. J Reprod Fertil 1985; 74:709–716.
De Schepper GG, van Noorden CJ, Houtkooper JM . Age-related changes of glucose-6-phosphate dehydrogenase activity in mouse oocytes. Histochem J 1987; 19:467–470.
Miao Y, Ma S, Liu X, et al. Fate of the first polar bodies in mouse oocytes. Mol Reprod Dev 2004; 69:66–76.
Liu XY, Mal SF, Miao DQ, Liu DJ, Bao S, Tan JH . Cortical granules behave differently in mouse oocytes matured under different conditions. Hum Reprod 2005; 20:3402–3413.
Funahashi H, Cantley TC, Stumpf TT, Terlouw SL, Day BN . Use of low-salt culture medium for in vitro maturation of porcine oocytes is associated with elevated oocyte glutathione levels and enhanced male pronuclear formation after in vitro fertilization. Biol Reprod 1994; 51:633–639.
Chatot CL, Ziomek CA, Bavister BD, Lewis JL, Torres I . An improved culture medium supports development of random-breed 1-cell mouse embryos in vitro. J Reprod Fertil 1989; 86:679–688.
Mattson BA, Albertini DF . Oogenesis: chromatin and microtubule dynamics during meiotic prophase. Mol Reprod Dev 1990; 25:374–383.
Liu J, Yagi T, Sadamori H, et al. Annexin V assay-proven anti-apoptotic effect of ascorbic acid 2-glucoside after cold ischemia/reperfusion injury in rat liver transplantation. Acta Med Okayama 2003; 57:209–216.
Rodriguez-Gonzalez E, Lopez-Bejar M, Izquierdo D, Paramio MT . Developmental competence of prepubertal goat oocytes selected with brilliant cresyl blue and matured with cysteamine supplementation. Reprod Nutr Dev 2003; 43:179–187.
de Matos DG, Furnus CC, Moses DF, Martinez AG, Matkovic M . Stimulation of glutathione synthesis of in vitro matured bovine oocytes and its effect on embryo development and freezability. Mol Reprod Dev 1996; 45:451–457.
de Matos DG, Furnus CC . The importance of having high glutathione (GSH) level after bovine in vitro maturation on embryo development effect of beta-mercaptoethanol, cysteine and cystine. Theriogenology 2000; 53:761–771.
Abeydeera LR, Wang WH, Cantley TC, Prather RS, Day BN . Presence of beta-mercaptoethanol can increase the glutathione content of pig oocytes matured in vitro and the rate of blastocyst development after in vitro fertilization. Theriogenology 1998; 50:747–756.
Perreault SD, Barbee RR, Slott VL . Importance of glutathione in the acquisition and maintenance of sperm nuclear decondensing activity in maturing hamster oocytes. Dev Biol 1988; 125:181–186.
Yoshida M, Ishigaki K, Nagai T, Chikyu M, Pursel VG . Glutathione concentration during maturation and after fertilization in pig oocytes: relevance to the ability of oocytes to form male pronucleus. Biol Reprod 1993; 49:89–94.
Cognie Y, Baril G, Poulin N, Mermillod P . Current status of embryo technologies in sheep and goat. Theriogenology 2003; 59:171–188.
de Matos DG, Gasparrini B, Pasqualini SR, Thompson JG . Effect of glutathione synthesis stimulation during in vitro maturation of ovine oocytes on embryo development and intracellular peroxide content. Theriogenology 2002; 57:1443–1451.
Nishi Y, Takeshita T, Sato K, Araki T . Change of the mitochondrial distribution in mouse ooplasm during in vitro maturation. J Nippon Med Sch 2003; 70:408–415.
Miki H, Ogonuki N, Inoue K, Baba T, Ogura A . Improvement of cumulus-free oocyte maturation in vitro and its application to microinsemination with primary spermatocytes in mice. J Reprod Dev 2006; 52:239–248.
El Shourbagy SH, Spikings EC, Freitas M, St John JC . Mitochondria directly influence fertilisation outcome in the pig. Reproduction 2006; 131:233–245.
Fair T, Hyttel P, Greve T . Bovine oocyte diameter in relation to maturational competence and transcriptional activity. Mol Reprod Dev 1995; 42:437–442.
Otoi T, Yamamoto K, Koyama N, Tachikawa S, Suzuki T . Bovine oocyte diameter in relation to developmental competence. Theriogenology 1997; 48:769–774.
Raghu HM, Nandi S, Reddy SM . Follicle size and oocyte diameter in relation to developmental competence of buffalo oocytes in vitro. Reprod Fertil Dev 2002; 14:55–61.
Ikeda K, Takahashi Y . Comparison of maturational and developmental parameters of oocytes recovered from prepubertal and adult pigs. Reprod Fertil Dev 2003; 15:215–221.
Hyun SH, Lee GS, Kim DY, et al. Effect of maturation media and oocytes derived from sows or gilts on the development of cloned pig embryos. Theriogenology 2003; 59:1641–1649.
Zuccotti M, Rossi PG, Martinez A, Garagna S, Forabosco A, Redi CA . Meiotic and developmental competence of mouse antral oocytes. Biol Reprod 1998; 58:700–704.
Duby RT, Damiani P, Looney CR, Fissore RA, Robl JM . Prepubertal calves as oocyte donors: promises and problems. Theriogenology 1996; 45:121–130.
Gandolfi F, Milanesi E, Pocar P, et al. Comparative analysis of calf and cow oocytes during in vitro maturation. Mol Reprod Dev 1998; 49:168–175.
Blondin P, Sirard MA . Oocyte and follicular morphology as determining characteristics for developmental competence in bovine oocytes. Mol Reprod Dev 1995; 41:54–62.
Moor RM, Lee C, Dai YF, Fulka J Jr . Antral follicles confer developmental competence on oocytes. Zygote 1996; 4:289–293.
de Wit AA, Wurth YA, Kruip TA . Effect of ovarian phase and follicle quality on morphology and developmental capacity of the bovine cumulus-oocyte complex. J Anim Sci 2000; 78:1277–1283.
Han ZB, Lan GC, Wu YG, et al. Interactive effects of granulosa cell apoptosis, follicle size, COC morphology and cumulus expansion on the developmental competence of goat oocytes: a study using the well-in-drop (WID) culture system. Reproduction 2006; 132:749–758.
Senbon S, Hirao Y, Miyano T . Interactions between the oocyte and surrounding somatic cells in follicular development: lessons from in vitro culture. J Reprod Dev 2003; 49:259–269.
Acknowledgements
This study was supported by grants from the China National Natural Science Foundation (Nos. 30430530 and 30571337) and from the Momentous Research Project of the China Ministry of Science and Technology (No. 2006CB944003).
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Wu, YG., Liu, Y., Zhou, P. et al. Selection of oocytes for in vitro maturation by brilliant cresyl blue staining: a study using the mouse model. Cell Res 17, 722–731 (2007). https://doi.org/10.1038/cr.2007.66
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DOI: https://doi.org/10.1038/cr.2007.66
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