Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Reproductive Biology and Endocrinology
Published in: Reproductive Biology and Endocrinology 1/2017

Open Access 01-12-2017 | Research

Oocyte polarized light microscopy, assay of specific follicular fluid metabolites, and gene expression in cumulus cells as different approaches to predict fertilization efficiency after ICSI

Authors: Alberto Revelli, Stefano Canosa, Loredana Bergandi, Oleksii A. Skorokhod, Valentina Biasoni, Andrea Carosso, Angela Bertagna, Milena Maule, Elisabetta Aldieri, Maria Diletta D’Eufemia, Francesca Evangelista, Nicola Colacurci, Chiara Benedetto

Published in: Reproductive Biology and Endocrinology | Issue 1/2017

Login to get access

Abstract

Background

The complex relationship between oocyte morphology, specific follicular fluid metabolites, gene expression in cumulus granulosa cells, and oocyte competence toward fertilization and embryo development still needs further clarification.

Methods

Forty-six oocytes retrieved from the largest pre-ovulatory follicle of patients undergoing intra-cytoplasmic sperm injection (ICSI) were considered assessing: (a) oocyte morphological characteristics at polarized light microscopy (PLM), (b) specific follicular fluid (FF) metabolites previously suggested to influence oocyte competence (AMH, markers of redox status and of cytotoxicity), (c) transcription of AMH and AMH type II receptor genes in cumulus cells. Data were analyzed using mono-parametric tests and multivariable logistic analysis in order to correlate morphological and biochemical data with fertilization.

Results

Comparing normally fertilized oocytes (n = 29, F group) with unfertilized (n = 17, nF group) we observed that: (a) the meiotic spindle area and major axis were significantly higher in nF group and in fertilized oocytes undergoing an early embryo development arrest; (b) AMH level in FF was comparable in F and nF groups; (c) the FF of nF group contained significantly higher levels of cytotoxicity (lactate dehydrogenase) and oxidative stress (Cu,Zn-superoxide dismutase, catalase, 4-hydroxynonenal-protein conjugates) markers; (d) cumulus cells of nF group showed significantly higher AMH receptor type II gene expression.

Conclusions

Taken together, these observations suggest that an excessive cytotoxicity level can alter AMH signal transduction within cumulus cells, in turn leading to partial inhibition of aromatase activity, altered cytoplasmic maturation and increased oxidative stress, factors able to impair oocyte fertilization competence and embryo growth.
Appendix
Available only for authorised users
Literature
1.
2.
Rienzi L, Vajta G, Ubaldi F. Predictive value of oocyte morphology in human IVF: a systematic review of the literature. Hum Reprod. 2011;17:34–45.CrossRef
3.
Montag M, Köster M, van der Ven K, van der Ven H. Gamete competence assessment by polarizing optics in assisted reproduction. Hum Reprod Update. 2011;17:654–66.CrossRefPubMed
4.
Madaschi C, de Souza BTC, de Almeida Ferreira Braga DP, Pasqualotto FF, Iaconelli A Jr, Borges E Jr. Spindle imaging: a marker for embryo development and implantation. Fertil Steril. 2008;90:194–8.CrossRefPubMed
5.
Braga DP, Figueira Rde C, Rodrigues D, Madaschi C, Pasqualotto FF, Iaconelli A Jr, et al. Prognostic value of meiotic spindle imaging on fertilization rate and embryo development in in vitro-matured human oocytes. Fertil Steril. 2008;90:429–33.CrossRefPubMed
6.
Montag M, van der Ven H. Symposium: innovative techniques in human embryo viability assessment. Oocyte assessment and embryo viability prediction: birefringence imaging. Reprod BioMed Online. 2008;17:454–60.CrossRefPubMed
7.
Rama Raju GA, Prakash GJ, Krishna KM, Madan K. Meiotic spindle and zona pellucida characteristics as predictors of embryonic development: a preliminary study using PolScope imaging. Reprod BioMed Online. 2007;14:166–74.CrossRefPubMed
8.
Ebner T, Balaban B, Moser M, Shebl O, Urman B, Ata B, et al. Automatic user-independent zona pellucida imaging at the oocyte stage allows for the prediction of preimplantation development. Fertil Steril. 2010;94:913–20.CrossRefPubMed
9.
Shen Y, Stalf T, Mehnert C, Eichenlaub-Ritter U, Tinneberg HR. High magnitude of light retardation by the zona pellucida is associated with conception cycles. Hum Reprod. 2005;20:1596–606.CrossRefPubMed
10.
Molinari E, Evangelista F, Racca C, Cagnazzo C, Revelli A. Polarized light microscopy-detectable structures of human oocytes and embryos are related to the likelihood of conception in IVF. J Assist Reprod Genet. 2012;29:1117–22.CrossRefPubMedPubMedCentral
11.
Revelli A, Delle Piane L, Casano S, Molinari E, Massobrio M, Rinaudo P. Follicular fluid content and oocyte quality: from single biochemical markers to metabolomics. Reprod Biol Endocrinol. 2009;7:40.CrossRefPubMedPubMedCentral
12.
Takahashi C, Fujito A, Kazuka M, Sugiyama R, Ito H, Isaka K. Anti-Müllerian hormone substance from follicular fluid is positively associated with success in oocyte fertilization during in vitro fertilization. Fertil Steril. 2008;89:586–91.CrossRefPubMed
13.
Kim JH, Lee JR, Chang HJ, Jee BC, Suh CS, Kim SH. Anti-Müllerian hormone levels in the follicular fluid of the preovulatory follicle: a predictor for oocyte fertilization and quality of embryo. J Korean Med Sci. 2014;29:1266–70.CrossRefPubMedPubMedCentral
14.
Jozwik M, Wolczynski S, Jozwik M, Szamatowicz M. Oxidative stress markers in preovulatory follicular fluid in humans. Mol Hum Reprod. 1999;5:409–13.CrossRefPubMed
15.
Oyawoye O, Abdel Gadir A, Garner A, Constantinovici N, Perrett C, Hardiman P. Antioxidants and reactive oxygen species in follicular fluid of women undergoing IVF: relationship to outcome. Hum Reprod. 2003;18:2270–4.CrossRefPubMed
16.
Pasqualotto EB, Agarwal A, Sharma RK, Izzo VM, Pinotti JA, Joshi NJ, et al. Effect of oxidative stress in follicular fluid on the outcome of assisted reproductive procedures. Fertil Steril. 2004;81:973–6.CrossRefPubMed
17.
Das S, Chattopadhyay R, Ghosh S, Ghosh S, Goswami SK, Chakravarty BN, et al. Reactive oxygen species level in follicular fluid--embryo quality marker in IVF? Hum Reprod. 2006;21:2403–7.CrossRefPubMed
18.
Scalici E, Traver S, Molinari N, Mullet T, Monforte M, Vintejoux E, et al. Cell-free DNA in human follicular fluid as a biomarker of embryo quality. Hum Reprod. 2014;29:2661–9.CrossRefPubMed
19.
Huber M, Hadziosmanovic N, Berglund L, Holte J. Using the ovarian sensitivity index to define poor, normal, and high response after controlled ovarian hyperstimulation in the long gonadotropin-releasing hormone-agonist protocol: suggestions for a new principle to solve an old problem. Fertil Steril. 2013;100:1270–6.CrossRefPubMed
20.
World Health Organization. Laboratory manual for the examination and processing of human semen. 2010. Fifth edition.
21.
Holte J, Berglund L, Milton K, Garello C, Gennarelli G, Revelli A, et al. Construction of an evidence-based integrated morphology cleavage embryo score for implantation potential of embryos scored and transferred on day 2 after oocyte retrieval. Hum Reprod. 2007;22(2):548–57.CrossRefPubMed
22.
Alpha scientists in reproductive medicine and ESHRE special interest Group of Embryology. The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting. Hum Reprod. 2011;26(6):1270–83.CrossRef
23.
Beutler E. Red cell metabolism: a manual of biochemical methods. New York: Grune & Stratton; 1971. p. 56–68.
24.
Goldstein S, Squadrito GL, Pryor WA, Czapski G. Direct and indirect oxidations by peroxynitrite, neither involving the hydroxyl radical. Free Radic Biol Med. 1996;21:965–74.CrossRefPubMed
25.
Nebot C, Moutet M, Huet P, Xu JZ, Yadan JC, Chaudiere J. Spectrophotometric assay of superoxide dismutase activity based on the activated autooxidation of a tetracyclic catechol. Anal Biochem. 1993;214:442–51.CrossRefPubMed
26.
Carbone MC, Tatone C, Delle Monache S, Marci R, Caserta D, Colonna R, et al. Antioxidant enzymatic defences in human follicular fluid: characterization and age-dependent changes. Mol Hum Reprod. 2003;9:639–43.CrossRefPubMed
27.
Skorokhod OA, Davalos-Schafler D, Gallo V, Valente E, Ulliers D, Notarpietro A, et al. Oxidative stress-mediated antimalarial activity of plakortin, a natural endoperoxide from the tropical sponge Plakortis simplex. Free Radic Biol Med. 2015;89:624–37.CrossRefPubMed
28.
Skorokhod OA, Barrera V, Heller R, Carta F, Turrini F, Arese P, et al. Malarial pigment hemozoin impairs chemotactic motility and transendothelial migration of monocytes via 4-hydroxynonenal. Free Radic Biol Med. 2014;75:210–21.CrossRefPubMed
29.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 2001;25:402–8.CrossRefPubMed
30.
Korkmaz C, Cinar O, Akyol M. The relationship between meiotic spindle imaging and outcome of intracytoplasmic sperm injection: a retrospective study. Gynecol Endocrinol. 2011;27:737–41.CrossRefPubMed
31.
Howe K, FitzHarris G. Recent insights into spindle function in mammalian oocytes and early embryos. Biol Reprod. 2013;89:1–9.CrossRef
32.
Schatten H, Sun QY. Centrosome and microtubule functions and dysfunctions in meiosis: implications for age-related infertility and developmental disorders. Reprod Fertil Dev. 2015;27:934–43.CrossRefPubMed
33.
La Marca A, Sighinolfi G, Radi D, Argento C, Baraldi E, Artenisio AC, et al. Anti-Mullerian hormone (AMH) as a predictive marker in assisted reproductive technology (ART). Hum Reprod Update. 2010;16:113–30.CrossRefPubMed
34.
Seifer DB, MacLaughlin DT, Christian BP, Feng B, Shelden RM. Early follicular serum müllerian-inhibiting substance levels are associated with ovarian response during assisted reproductive technology cycles. Fertil Steril. 2002;77:468–71.
35.
Fanchin R, Mendez Lozano DH, Frydman N, Gougeon A, di Clemente N, Frydman R, et al. Anti-Müllerian hormone concentrations in the follicular fluid of the preovulatory follicle are predictive of the implantation potential of the ensuing embryo obtained by in vitro fertilization. J Clin Endocrinol Metab. 2007;92:1796–802.
36.
Asimakopoulos B, Abu-Hassan D, Metzen E, Al-Hasani S, Diedrich K, Nikolettos N. The levels of steroid hormones and cytokines in individual follicles are not associated with the fertilization outcome after intracytoplasmic sperm injection. Fertil Steril. 2008;90:60–4.CrossRefPubMed
37.
Mashiach R, Amit A, Hasson J, Amzalzg S, Almog B, Ben-Yosef D, et al. Follicular fluid levels of anti-Mullerian hormone as a predictor of oocyte maturation, fertilization rate, and embryonic development in patients with polycystic ovary syndrome. Fertil Steril. 2010;93:2299–302.CrossRefPubMed
38.
Mehta BN, Chimote MN, Chimote NN, Nath NM, Chimote NM. Follicular-fluid anti-Mullerian hormone (FF AMH) is a plausible biochemical indicator of functional viability of oocyte in conventional in vitro fertilization (IVF) cycles. J Hum Reprod Sci. 2013;6:99–105.
39.
Tramišak Milaković T, Panić Horvat L, Čavlović K, Smiljan Severinski N, Vlašić H, Vlastelić I, et al. Follicular fluid anti-Müllerian hormone: a predictive marker of fertilization capacity of MII oocytes. Arch Gynecol Obstet. 2015;291:681–7.CrossRefPubMed
40.
Sikka SC. Role of oxidative stress and antioxidants in andrology and assisted reproductive technology. J Androl. 2004;25:5–18.CrossRefPubMed
41.
Pacella L, Zander-Fox DL, Armstrong DT, Lane M. Women with reduced ovarian reserve or advanced maternal age have an altered follicular environment. Fertil Steril. 2012;98:986–94.CrossRefPubMed
42.
Schwarzer E, Arese P, Skorokhod OA. Role of the lipoperoxidation product 4-hydroxynonenal in the pathogenesis of severe malaria anemia and malaria immunodepression. Oxid Med Cell Longev. 2015;638416:11.
43.
Bergandi L, Basso G, Evangelista F, Canosa S, Dalmasso P, Aldieri E, et al. Inducible nitric oxide synthase and heme oxygenase 1 are expressed in human cumulus cells and may be used as biomarkers of oocyte competence. Reprod Sci. 2014;21:1370–7.CrossRefPubMed
44.
Fotakis G, Timbrell JA. In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett. 2006;160:171–7.CrossRefPubMed
45.
Segev DL, Hoshiya Y, Hoshiya M, Tran TT, Carey JL, Stephen AE, et al. Mullerian-inhibiting substance regulates NF-kappa B signaling in the prostate in vitro and in vivo. Proc Natl Acad Sci U S A. 2002 Jan 8;99(1):239–44.CrossRefPubMedPubMedCentral
Metadata
Title
Oocyte polarized light microscopy, assay of specific follicular fluid metabolites, and gene expression in cumulus cells as different approaches to predict fertilization efficiency after ICSI
Authors
Alberto Revelli
Stefano Canosa
Loredana Bergandi
Oleksii A. Skorokhod
Valentina Biasoni
Andrea Carosso
Angela Bertagna
Milena Maule
Elisabetta Aldieri
Maria Diletta D’Eufemia
Francesca Evangelista
Nicola Colacurci
Chiara Benedetto
Publication date
01-12-2017
Publisher
BioMed Central
Published in
Reproductive Biology and Endocrinology / Issue 1/2017
Electronic ISSN: 1477-7827
DOI
https://doi.org/10.1186/s12958-017-0265-2

Other articles of this Issue 1/2017

Reproductive Biology and Endocrinology 1/2017 Go to the issue

Short Communication

Oleic acid induces down-regulation of the granulosa cell identity marker FOXL2, and up-regulation of the Sertoli cell marker SOX9 in bovine granulosa cells

Research

A new insight into male fertility preservation for patients with completely immotile spermatozoa

Review

Risks, benefits size and clinical implications of combined oral contraceptive use in women with polycystic ovary syndrome

Research

Towards a better understanding of preimplantation genetic screening for aneuploidy: insights from a virtual trial for women under the age of 40 when transferring embryos one at a time

Review

Role of microRNAs in premature ovarian insufficiency

Review

5α-reductase activity in women with polycystic ovary syndrome: a systematic review and meta-analysis