title: Fertilization ability of porcine oocytes reconstructed from ooplasmic fragments produced and characterized after serial centrifugations.
abstract: Mitochondria are reported to be critical in in vitro maturation of oocytes and subsequent embryo development after fertilization, but their contribution for fertilization has not been investigated in detail. In the present study, we investigate the contribution of mitochondria to fertilization using reconstructed porcine oocytes by fusion of ooplasmic fragments produced by serial centrifugations (centri-fusion). Firstly, we evaluated the characteristics of ooplasmic fragments. Three types of fragments were obtained by centrifugation of porcine oocytes matured in vitro for 46 h: brownish (B), transparent (T) and large (L) fragments containing both B and T parts in a fragment. The production efficiencies of these types of fragments were 71.7, 91.0 and 17.8 fragments/100 oocytes, respectively. In experiments, L fragments were excluded because they contained both brownish and transparent components that were apparently intermediate between B and T fragments. Observations by confocal microscopy after staining with MitoTracker Red CMXRos® and transmission electron microscopy revealed highly condensed active mitochondria in B fragments in contrast to T fragments that contained only sparse organelles. We reconstructed oocytes by fusion of a karyoplast and two cytoplasts from B and T fragments (B and T oocytes, respectively). The B oocytes showed higher sperm penetration (95.8%) and male pronuclear formation rates (94.2%) by in vitro fertilization than T oocytes (66.7% and 50.0%, respectively). These results suggest that the active mitochondria in oocytes may be related to their ability for fertilization.
authors: Animal Resource Science Center, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Ibaraki 319-0206, Japan.
authors: Nguyen
authors: N
authors: Viet Linh
authors: Kazuhiro
authors: K
authors: Kikuchi
authors: Michiko
authors: M
authors: Nakai
authors: Fuminori
authors: F
authors: Tanihara
authors: Junko
authors: J
authors: Noguchi
authors: Hiroyuki
authors: H
authors: Kaneko
authors: Thanh Quang
authors: TQ
authors: Dang-Nguyen
authors: Nguyen Thi
authors: NT
authors: Men
authors: Nguyen
authors: N
authors: Van Hanh
authors: Tamas
authors: T
authors: Somfai
authors: Bui Xuan
authors: BX
authors: Nguyen
authors: Takashi
authors: T
authors: Nagai
authors: Noboru
authors: N
authors: Manabe
mesh_terms: D000003:Abattoirs; D000818:Animals; D017849:Animals, Inbred Strains; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D003433:Crosses, Genetic; D015925:Cryopreservation; D022083:Cytoplasmic Structures / Q000502:physiology / Q000648:ultrastructure; D055664:Electrochemical Techniques; D005260:Female; D005307:Fertilization in Vitro; D059471:In Vitro Oocyte Maturation Techniques; D007564:Japan; D008297:Male; D008561:Membrane Fusion; D008928:Mitochondria / Q000502:physiology* / Q000648:ultrastructure; D009865:Oocytes / Q000502:physiology* / Q000648:ultrastructure; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa; D034421:Sus scrofa; D015854:Up-Regulation
references: Mitochondrion. 2011 Sep;11(5):797-813
references: 20933103
references: Biol Reprod. 2001 Mar;64(3):904-9
references: 11207207
references: Rev Reprod. 1996 Jan;1(1):33-9
references: 9414436
references: Nature. 1995 Mar 30;374(6521):416
references: 7700349
references: Reproduction. 2001 Jul;122(1):155-63
references: 11425340
references: Eur J Histochem. 2004 Oct-Dec;48(4):347-56
references: 15718200
references: Mol Hum Reprod. 2007 Nov;13(11):759-70
references: 17893093
references: J Reprod Fertil Suppl. 1993;48:61-73
references: 8145215
references: Mol Reprod Dev. 2001 Dec;60(4):481-90
references: 11746959
references: Semin Cell Dev Biol. 2006 Apr;17(2):314-23
references: 16574440
references: Biol Reprod. 2009 Mar;80(3):563-71
references: 19038857
references: Theriogenology. 2004 Jun;61(9):1675-89
references: 15019463
references: Mitochondrion. 2010 Mar;10(2):137-42
references: 20005304
references: Chem Res Toxicol. 2011 Oct 17;24(10):1630-2
references: 21950265
references: Aust J Biol Sci. 1973 Jun;26(3):653-67
references: 4795241
references: Biol Reprod. 2002 Apr;66(4):1033-41
references: 11906923
references: Am J Anat. 1984 Nov;171(3):335-55
references: 6517035
references: Development. 2004 Jul;131(13):3057-67
references: 15163630
references: Hum Reprod. 1996 Jul;11(7):1499-503
references: 8671492
references: Biol Reprod. 1993 Dec;49(6):1362-8
references: 8286618
references: Semin Cell Dev Biol. 2009 May;20(3):346-53
references: 19530278
references: Biol Reprod. 2005 Jun;72(6):1397-404
references: 15716395
references: Anim Reprod Sci. 2007 Mar;98(1-2):72-96
references: 17110061
references: Biol Reprod. 2007 May;76(5):900-5
references: 17267701
references: Theriogenology. 1998 Sep;50(4):615-23
references: 10732152
references: Biol Reprod. 2001 Dec;65(6):1648-54
references: 11717124
references: Reprod Domest Anim. 2010 Aug;45(4):659-65
references: 19144027
references: Gamete Res. 1987 Sep;18(1):67-76
references: 3507363
references: Reprod Fertil Dev. 2012;24(5):743-52
references: 22697124
references: Reproduction. 2006 Feb;131(2):233-45
references: 16452717
references: Biol Reprod. 2005 May;72(5):1218-23
references: 15659704
references: Hum Reprod. 1990 Apr;5(3):319-22
references: 2351715
references: Dev Genet. 1995;16(1):36-43
references: 7538924
references: Dev Biol. 1993 Jul;158(1):62-78
references: 8392472
references: Mol Reprod Dev. 2000 Mar;55(3):256-64
references: 10657044
references: J Reprod Dev. 2011 Oct;57(5):620-6
references: 21685709
references: Hum Reprod. 2001 May;16(5):909-17
references: 11331637
references: Reproduction. 2006 Mar;131(3):603-11
references: 16514203
references: Dev Biol. 1999 Jul 15;211(2):157-76
references: 10395780
references: J Cell Physiol. 2010 Sep;224(3):672-80
references: 20578238
references: Hum Reprod. 2003 Nov;18(11):2429-40
references: 14585897
references: Cloning Stem Cells. 2007 Summer;9(2):216-28
references: 17579554
references: Int J Androl. 2002 Apr;25(2):84-93
references: 11903657