Porcine Oocytes with Meiotic Competence are Synchronized Early Stage of Germinal Vesicle (GV)

  • Park, Kwang-Wook (Dept. of Animal Science & Technology, Sunchon National University)
  • 투고 : 2010.08.26
  • 심사 : 2010.09.15
  • 발행 : 2010.09.30

초록

Correlations between cumulus cells and germinal vesicle (GV) chromatin configuration were examined in porcine oocytes. Cumulus-oocyte complexes (COCs) were collected from 2~6 mm follicles and divided into three categories according to cumulus cell morphology. "A" group was compacted COCs with more than three cumulus cell layers. "B" group was COCs with less cumulus cell layers than "A" group. "C" group was COCs with one or less layer of cumulus cells. Cumulus cells were removed 0.1% hyaluronidase, and denuded oocytes were stained with Hoechst 33342. GV chromatin configuration was classified into GV-Con and GV-Dis. GV-Con meant that a nucleus was surrounded by condensed chromatin in a ring. GV-Dis meant that filamentous chromatin clumps were distributed in nucleus. The proportion (80.2%) of GV-Con in "A" group was significantly higher than "B" (62.0%) or "C" (44.9%). The proportion (55.1%) of GV-Dis in "C" group was significantly higher than "A" (19.8%) or "B" (38.0%). The meiotic competence of COCs was examined after 44 h culture. The proportion (90.0%) of oocytes reaching to metaphase II (M-II) in "A" group was significantly higher than "B" (76.5%) or "C" (45.5%). In conclusion, oocytes with good quality cumulus cell layers are synchronized early GV stage, and early GV stage is important for meiotic competence in pigs.

키워드

참고문헌

  1. Chian RC, Sirard MA (1995): Effects of cumulus cells and follicle-stimulating hormone during in vitro maturation on parthenogenetic activation of bovine oocytes. Mol Reprod Dev 42:425-431. https://doi.org/10.1002/mrd.1080420408
  2. Chohan KR, Hunter AG (2003): Meiotic competence of bovine fetal oocytes following in vitro maturation. Anim Reprod Sci 76:43-51. https://doi.org/10.1016/S0378-4320(02)00235-X
  3. Funahashi H, Cantley TC, Day BN (1997a): Preincubation of cumulus-ooycte complexes before exposure to gonadotropins improves the developmental competence of porcine embryos matured and fertilized in vitro. Theriogenology 47:679-686. https://doi.org/10.1016/S0093-691X(97)00026-5
  4. Funahashi H, Cantley TC, Day BN (1997b): Synchronization of meiosis in porcine oocytes by exposure to dibutyryl cyclic adenosine monophosphate improves developmental competence following in vitro fertilization. Biol Reprod 57:49-53. https://doi.org/10.1095/biolreprod57.1.49
  5. Funahashi H, Day BN (1997): Advances in in vitro production of pig embryos. J Reprod Fertil Suppl. 52:271-283.
  6. Galeati G, Modina S, Lauria A, Mattioli M (1991): Follicle somatic cells influence pig oocyte penetrability and cortical granule distribution. Mol Reprod Dev 29:40-46. https://doi.org/10.1002/mrd.1080290107
  7. Geshi M, Takenouchi N, Yamauchi N, Nagai T (2000): Effects of sodium pyruvate in nonserum maturation medium on maturation. fertilization, and subsequent development of bovine oocytes with or without cumulus cells. Biol Reprod 63:1730-1734. https://doi.org/10.1095/biolreprod63.6.1730
  8. Grupen CG, Nagashima H, Nottle MB (1997): Asynchronous meiotic progression in porcine oocytes matured in vitro: a cause of polyspermic fertilization? Reprod Fertil Dev 9:187-191. https://doi.org/10.1071/R96087
  9. Ka HH, Sawai K, Wang WH, Im KS, Niwa K (1997): Amino acids in maturation medium and presence of cumulus cells at fertilization promote male pronuclear formation in porcine oocytes matured and penetrated in vitro. Biol Reprod 57:1478-1483. https://doi.org/10.1095/biolreprod57.6.1478
  10. Khurana NK, Niemann H (2000): Effects of oocyte quality, oxygen tension. embryo density, cumulus cells and energy substrates on cleavage and morula/blastocyst formation of bovine embryos. Theriogenology 54:741-756. https://doi.org/10.1016/S0093-691X(00)00387-3
  11. Kikuchi K, Nagai T, Motlik J, Shioya Y, Izaike Y (1993): Effect of follicle cells on in vitro fertilization of pig follicular oocytes. Theriogenology 39:593-599. https://doi.org/10.1016/0093-691X(93)90246-2
  12. Long CR, Dobrinsky JR, Johnson LA (1997): In vitro production of pig embryos. comparisons of culture media and boars: Theriogenology 51:1375-1390.
  13. Mattioli M, Galeati G, Bacci ML, Seren E (1988a): Follicular factors influence oocyte fertilizability by modulating the intercellular cooperation between cumulus cells and oocyte. Gamete Res 21:223-232. https://doi.org/10.1002/mrd.1120210304
  14. Mattioli M, Galeati G, Seren E (1988b): Effect of follicle somatic cells during pig oocyte maturation on egg penetrability and male pronucleus formation. Gamete Res 20:177-183. https://doi.org/10.1002/mrd.1120200208
  15. Moor RM, Mattioli M, Ding J, Nagai T (1990): Maturation of pig oocytes in vivo and in vitro. J Reprod Fertil Suppl 40:197-210.
  16. MotliK J, Fulka J (1976): Breakdown of the germinal vesicle in pig oocytes in vivo and in vitro. J Exp Zool 198:155-162. https://doi.org/10.1002/jez.1401980205
  17. Nagai T, Ebihara M, Onishi A, Kubo M (1997): Germinal vesicle stages in pig follicular oocytes collected by different methods. J Reprod Deve 43:339-343. https://doi.org/10.1262/jrd.43.339
  18. Ozawa M, Nagai T, Somfai T, Nakai M, Maedomari N, Miyazaki H, Kaneko H, Noguchi J, Kikuchi K (2010): Cumulus cell-enclosed oocytes acquire a capacity to synthesize GSH by FSH stimulation during in vitro maturation in pigs. J Cell Physiol 222:294-301. https://doi.org/10.1002/jcp.21949
  19. Park KW, Funahashi H, Niwa K (1999): Co-culture of cumulus-enclosed bovine oocytes with theca cells induces the meiotic arrest but does not inhibit germinal vesicle development. J Reprod Dev 45:223-231. https://doi.org/10.1262/jrd.45.223
  20. Sun XS, Liu Y, Yue KZ, Ma SF, Tan JH (2004): Changes in germinal vesicle (GV) chromatin configurations during growth and maturation of porcine oocytes. Mol Reprod Dev 69:228-234. https://doi.org/10.1002/mrd.20123
  21. Tsafriri A, Channing CP (1975): Influence of follicular maturation and culture conditions on the meiosis of pig oocytes in vitro. J Reprod Fertil 43:149-152. https://doi.org/10.1530/jrf.0.0430149
  22. Xia GL, Kikuchi K, Noguchi J, Izaike Y (2000): Short time priming of pig cumulus-oocyte complexes with FSH and forskolin in the presence of hypoxanthine stimulates cumulus cells to secrete a meiosis-activating substance. Theriogenology 53:1807-1815. https://doi.org/10.1016/S0093-691X(00)00316-2