The morphological assessment of oocytes is important for embryologists to identify and select MII oocytes in IVF/ICSI cycles. Dysmorphism of oocytes decreases viability and the developmental potential of oocytes as well as the clinical pregnancy rate. Several reports have suggested that oocytes with a dark zona pellucida (DZP) correlate with the outcome of IVF treatment. However, the effect of DZP on oocyte quality, fertilization, implantation, and pregnancy outcome were not investigated in detail. In this study, a retrospective analysis was performed in 268 infertile patients with fallopian tube obstruction and/or male factor infertility. In 204 of these patients, all oocytes were surrounded by a normal zona pellucida (NZP, control group), whereas 46 patients were found to have part of their retrieved oocytes enclosed by NZP and the other by DZP (Group A). In addition, all oocytes enclosed by DZP were retrieved from 18 patients (Group B). No differences were detected between the control and group A. Compared to the control group, the rates of fertilization, good quality embryos, implantation and clinical pregnancy were significantly decreased in group B. Furthermore, mitochondria in oocytes with a DZP in both of the two study groups (A and B) were severely damaged with several ultrastructural alterations, which were associated with an increased density of the zona pellucida and vacuolization. Briefly, oocytes with a DZP affected the clinical outcome in IVF/ICSI cycles and appeared to contain more ultrastructural alterations. Thus, DZP could be used as a potential selective marker for embryologists during daily laboratory work.
References
[1]
Rienzi L, Ubaldi FM, Iacobelli M, Minasi MG, Romano S, et al. (2008) Significance of metaphase II human oocyte morphology on ICSI outcome. Fertil Steril 90: 1692–1700. doi: 10.1016/j.fertnstert.2007.09.024
[2]
Montag M, Schimming T, Koster M, Zhou C, Dorn C, et al. (2008) Oocyte zona birefringence intensity is associated with embryonic implantation potential in ICSI cycles. Reprod Biomed Online 16: 239–244. doi: 10.1016/s1472-6483(10)60580-9
[3]
Ebner T, Shebl O, Moser M, Sommergruber M, Tews G (2008) Developmental fate of ovoid oocytes. Hum Reprod 23: 62–66. doi: 10.1093/humrep/dem280
[4]
Ten J, Mendiola J, Vioque J, de Juan J, Bernabeu R (2007) Donor oocyte dysmorphisms and their influence on fertilization and embryo quality. Reprod Biomed Online 14: 40–48. doi: 10.1016/s1472-6483(10)60762-6
[5]
Otsuki J, Nagai Y, Chiba K (2007) Lipofuscin bodies in human oocytes as an indicator of oocyte quality. J Assist Reprod Genet 24: 263–270. doi: 10.1007/s10815-007-9130-0
[6]
Familiari G, Heyn R, Relucenti M, Nottola SA, Sathananthan AH (2006) Ultrastructural dynamics of human reproduction, from ovulation to fertilization and early embryo development. Int Rev Cytol 249: 53–141. doi: 10.1016/s0074-7696(06)49002-1
[7]
Verlhac MH, Lefebvre C, Guillaud P, Rassinier P, Maro B (2000) Asymmetric division in mouse oocytes: with or without Mos. Curr Biol 10: 1303–1306. doi: 10.1016/s0960-9822(00)00753-3
[8]
Xia P (1997) Intracytoplasmic sperm injection: correlation of oocyte grade based on polar body, perivitelline space and cytoplasmic inclusions with fertilization rate and embryo quality. Hum Reprod 12: 1750–1755. doi: 10.1093/humrep/12.8.1750
[9]
Mikkelsen AL, Lindenberg S (2001) Morphology of in-vitro matured oocytes: impact on fertility potential and embryo quality. Hum Reprod 16: 1714–1718. doi: 10.1093/humrep/16.8.1714
[10]
Miao YL, Kikuchi K, Sun QY, Schatten H (2009) Oocyte aging: cellular and molecular changes, developmental potential and reversal possibility. Hum Reprod Update 15: 573–585. doi: 10.1093/humupd/dmp014
[11]
Alikani M, Palermo G, Adler A, Bertoli M, Blake M, et al. (1995) Intracytoplasmic sperm injection in dysmorphic human oocytes. Zygote 3: 283–288. doi: 10.1017/s0967199400002707
Van Blerkom J, Henry G (1992) Oocyte dysmorphism and aneuploidy in meiotically mature human oocytes after ovarian stimulation. Hum Reprod 7: 379–390.
[14]
Shen Y, Stalf T, Mehnert C, Eichenlaub-Ritter U, Tinneberg HR (2005) High magnitude of light retardation by the zona pellucida is associated with conception cycles. Hum Reprod 20: 1596–1606. doi: 10.1093/humrep/deh811
[15]
Ebner T, Balaban B, Moser M, Shebl O, Urman B, et al. (2010) Automatic user-independent zona pellucida imaging at the oocyte stage allows for the prediction of preimplantation development. Fertil Steril 94: 913–920. doi: 10.1016/j.fertnstert.2009.03.106
[16]
Liu C, Litscher ES, Mortillo S, Sakai Y, Kinloch RA, et al. (1996) Targeted disruption of the mZP3 gene results in production of eggs lacking a zona pellucida and infertility in female mice. Proc Natl Acad Sci U S A 93: 5431–5436. doi: 10.1073/pnas.93.11.5431
[17]
Rankin TL, O’Brien M, Lee E, Wigglesworth K, Eppig J, et al. (2001) Defective zonae pellucidae in Zp2-null mice disrupt folliculogenesis, fertility and development. Development 128: 1119–1126.
[18]
Loret De MolaJR, Garside WT, Bucci J, Tureck RW, Heyner S (1997) Analysis of the human zona pellucida during culture: correlation with diagnosis and the preovulatory hormonal environment. J Assist Reprod Genet 14: 332–336. doi: 10.1007/bf02765837
[19]
Marco-Jimenez F, Naturil-Alfonso C, Jimenez-Trigos E, Lavara R, Vicente JS (2012) Influence of zona pellucida thickness on fertilization, embryo implantation and birth. Anim Reprod Sci 132: 96–100. doi: 10.1016/j.anireprosci.2012.04.008
[20]
Gabrielsen A, Lindenberg S, Petersen K (2001) The impact of the zona pellucida thickness variation of human embryos on pregnancy outcome in relation to suboptimal embryo development. A prospective randomized controlled study. Hum Reprod 16: 2166–2170. doi: 10.1093/humrep/16.10.2166
[21]
Palmstierna M, Murkes D, Csemiczky G, Andersson O, Wramsby H (1998) Zona pellucida thickness variation and occurrence of visible mononucleated blastomers in preembryos are associated with a high pregnancy rate in IVF treatment. J Assist Reprod Genet 15: 70–75. doi: 10.1007/bf02766828
[22]
Balaban B, Urman B (2006) Effect of oocyte morphology on embryo development and implantation. Reprod Biomed Online 12: 608–615. doi: 10.1016/s1472-6483(10)61187-x
[23]
Esfandiari N, Burjaq H, Gotlieb L, Casper RF (2006) Brown oocytes: implications for assisted reproductive technology. Fertil Steril 86: 1522–1525. doi: 10.1016/j.fertnstert.2006.03.056
[24]
Dozortsev D, De Sutter P, Dhont M (1994) Behaviour of spermatozoa in human oocytes displaying no or one pronucleus after intracytoplasmic sperm injection. Hum Reprod 9: 2139–2144.
[25]
Nottola SA, Coticchio G, Sciajno R, Gambardella A, Maione M, et al. (2009) Ultrastructural markers of quality in human mature oocytes vitrified using cryoleaf and cryoloop. Reprod Biomed Online 19 Suppl 317–27. doi: 10.1016/s1472-6483(10)60280-5
[26]
Liu DY, Baker HW (2000) Defective sperm-zona pellucida interaction: a major cause of failure of fertilization in clinical in-vitro fertilization. Hum Reprod 15: 702–708. doi: 10.1093/humrep/15.3.702
[27]
De Sutter P, Dozortsev D, Qian C, Dhont M (1996) Oocyte morphology does not correlate with fertilization rate and embryo quality after intracytoplasmic sperm injection. Hum Reprod 11: 595–597. doi: 10.1093/humrep/11.3.595
[28]
Nottola SA, Macchiarelli G, Coticchio G, Bianchi S, Cecconi S, et al. (2007) Ultrastructure of human mature oocytes after slow cooling cryopreservation using different sucrose concentrations. Hum Reprod 22: 1123–1133. doi: 10.1093/humrep/del463
[29]
Au HK, Yeh TS, Kao SH, Tzeng CR, Hsieh RH (2005) Abnormal mitochondrial structure in human unfertilized oocytes and arrested embryos. Ann N Y Acad Sci 1042: 177–185. doi: 10.1196/annals.1338.020
[30]
Motta PM, Nottola SA, Makabe S, Heyn R (2000) Mitochondrial morphology in human fetal and adult female germ cells. Hum Reprod 15 Suppl 2129–147. doi: 10.1093/humrep/15.suppl_2.129
[31]
Balaban RS, Nemoto S, Finkel T (2005) Mitochondria, oxidants, and aging. Cell 120: 483–495. doi: 10.1016/j.cell.2005.02.001
[32]
Dumollard R, Duchen M, Carroll J (2007) The role of mitochondrial function in the oocyte and embryo. Curr Top Dev Biol 77: 21–49. doi: 10.1016/s0070-2153(06)77002-8
[33]
Nagano M, Katagiri S, Takahashi Y (2006) ATP content and maturational/developmental ability of bovine oocytes with various cytoplasmic morphologies. Zygote 14: 299–304. doi: 10.1017/s0967199406003807
[34]
Eichenlaub-Ritter U, Wieczorek M, Luke S, Seidel T (2011) Age related changes in mitochondrial function and new approaches to study redox regulation in mammalian oocytes in response to age or maturation conditions. Mitochondrion 11: 783–796. doi: 10.1016/j.mito.2010.08.011
[35]
Hoodbhoy T, Dean J (2004) Insights into the molecular basis of sperm-egg recognition in mammals. Reproduction 127: 417–422. doi: 10.1530/rep.1.00181
[36]
Keefe D, Tran P, Pellegrini C, Oldenbourg R (1997) Polarized light microscopy and digital image processing identify a multilaminar structure of the hamster zona pellucida. Hum Reprod 12: 1250–1252. doi: 10.1093/humrep/12.6.1250
[37]
Van Blerkom J (2004) Mitochondria in human oogenesis and preimplantation embryogenesis: engines of metabolism, ionic regulation and developmental competence. Reproduction 128: 269–280. doi: 10.1530/rep.1.00240
[38]
Nottola SA, Makabe S, Stallone T, Familiari G, Correr S, et al. (2005) Surface morphology of the zona pellucida surrounding human blastocysts obtained after in vitro fertilization. Arch Histol Cytol 68: 133–141. doi: 10.1679/aohc.68.133
[39]
Motta PM, Nottola SA, Micara G, Familiari G (1988) Ultrastructure of human unfertilized oocytes and polyspermic embryos in an IVF-ET program. Ann N Y Acad Sci 541: 367–383. doi: 10.1111/j.1749-6632.1988.tb22274.x
[40]
Coticchio G, Borini A, Distratis V, Maione M, Scaravelli G, et al. (2010) Qualitative and morphometric analysis of the ultrastructure of human oocytes cryopreserved by two alternative slow cooling protocols. J Assist Reprod Genet 27: 131–140. doi: 10.1007/s10815-010-9394-7
[41]
El Shafie M, Sousa M, Windt M-L and Kruger TF. (2000) An Atlas of the Ultrastructure of Human Oocytes. Parthenon Publishing, New York, USA.
[42]
Sá R, Cunha M, Silva J, Luís A, Oliveira C, et al. (2011) Ultrastructure of tubular smooth endoplasmic reticulum aggregates in human metaphase II oocytes and clinical implications. Fertil Steril 96(1): 143–149. doi: 10.1016/j.fertnstert.2011.04.088
[43]
Silva RC, Báo SN, Jivago JL, Lucci CM (2011) Ultrastructural characterization of porcine oocytes and adjacent follicular cells during follicle development: lipid component evolution. Theriogenology 76(9): 1647–57. doi: 10.1016/j.theriogenology.2011.06.029
[44]
Windt ML, Coetzee K, Kruger TF, Marino H, Kitshoff MS, Sousa M, et al. (2001) Ultrastructural evaluation of recurrent and in-vitro maturation resistant metaphase I arrested oocytes. Hum Reprod 16(11): 2394–8.
[45]
Sousa M, da Silva JT, Silva J, Cunha M, Viana P, et al. (2013) Embryological, clinical and ultrastructural study of human oocytes presenting indented zona pellucida. Zygote 2: 1–13. doi: 10.1017/s0967199413000403
