Top

Journal of Assisted Reproduction and Genetics

Published in:

01-04-2017 | Reproductive Physiology and Disease

Extracellular microRNAs in follicular fluid and their potential association with oocyte fertilization and embryo quality: an exploratory study

Authors: Ronit Machtinger, Rodosthenis S. Rodosthenous, Michal Adir, Abdallah Mansour, Catherine Racowsky, Andrea A. Baccarelli, Russ Hauser

Published in: Journal of Assisted Reproduction and Genetics | Issue 4/2017

Abstract

Purpose

The purpose of this study is to determine the profile of extracellular microRNAs (exmiRNAs) in follicular fluid (FF) and explore their association with fertilization potential and embryo quality.

Methods

We collected FF from single follicles containing mature oocytes from 40 women undergoing IVF and we screened for the expression of 754 exmiRNAs in FF using the TaqMan OpenArray® qPCR platform. To determine the association of exmiRNAs and IVF outcomes, we compared their expression levels in FF samples that differ by fertilization status (normally, abnormally, and failed to fertilize) and embryo quality (top vs. non-top).

Results

We detected 207 exmiRNAs, of which miR-30d-5p, miR-320b, miR-10b-3p, miR-1291, and miR-720 were most prevalent. We identified four exmiRNAs with significant fold change (FC) when FF that contained normally fertilized was compared to failed to fertilize oocytes [miR-202-5p (FC = 1.82, p = 0.01), miR-206 (FC = 2.09, p = 0.04), miR-16-1-3p (FC = 1.88, p = 0.05), and miR-1244 (FC = 2.72, p = 0.05)]. We also found four exmiRNAs to be significantly differentially expressed in FF that yielded top quality versus non-top quality embryos [(miR-766-3p (FC = 1.95, p = 0.01), miR-663b (FC = 0.18, p = 0.02), miR-132-3p (FC = 2.45, p = 0.05), and miR-16-5p (FC = 3.80, p = 0.05)]. In-silico analysis revealed that several of these exmiRNAs are involved in pathways implicated in reproductive system diseases, organismal abnormalities, and organ development.

Conclusions

Our findings suggest that exmiRNAs in the follicular fluid can lead to downstream events that will affect fertilization and day 3 embryo morphology. We encourage further observational and experimental studies to confirm our findings and to determine the role of exmiRNAs in human reproduction.

Available only for authorised users

Eppig JJ. Oocyte control of ovarian follicular development and function in mammals. Reproduction. 2001;122(6):829–38.CrossRefPubMed

Eppig JJ, Chesnel F, Hirao Y, O’Brien MJ, Pendola FL, Watanabe S, et al. Oocyte control of granulosa cell development: how and why. Hum Reprod. 1997;12(11 Suppl):127–32.PubMed

Buccione R, Schroeder AC, Eppig JJ. Interactions between somatic cells and germ cells throughout mammalian oogenesis. Biol Reprod. 1990;43(4):543–7.CrossRefPubMed

Kidder GM, Vanderhyden BC. Bidirectional communication between oocytes and follicle cells: ensuring oocyte developmental competence. Can J Physiol Pharmacol. 2010;88(4):399–413. doi:10.1139/y10-009.CrossRefPubMedPubMedCentral

Winterhager E, Kidder GM. Gap junction connexins in female reproductive organs: implications for women’s reproductive health. Hum Reprod Update. 2015. doi:10.1093/humupd/dmv007.PubMed

Matzuk MM, Burns KH, Viveiros MM, Eppig JJ. Intercellular communication in the mammalian ovary: oocytes carry the conversation. Science. 2002;296(5576):2178–80. doi:10.1126/science.1071965.CrossRefPubMed

Yanez-Mo M, Siljander PR, Andreu Z, Zavec AB, Borras FE, Buzas EI, et al. Biological properties of extracellular vesicles and their physiological functions. J Extracellular Vesicles. 2015;4:27066. doi:10.3402/jev.v4.27066.CrossRef

Da Silveira JC, Sessions DR, Veeramachaneni DNR, Winger QA, Carnevale EM, Bouma GJ. MiRNAs within the ovarian follicle: Identification of cell-secreted vesicles as miRNA carriers. Biol Reprod. 2011;85(1).

Raposo G, Stoorvogel W. Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol. 2013;200(4):373–83. doi:10.1083/jcb.201211138.CrossRefPubMedPubMedCentral

10.

Zhang M, Ouyang H, Xia G. The signal pathway of gonadotrophins-induced mammalian oocyte meiotic resumption. Mol Hum Reprod. 2009;15(7):399–409. doi:10.1093/molehr/gap031.CrossRefPubMed

11.

Machtinger R, Laurent LC, Baccarelli AA. Extracellular vesicles: roles in gamete maturation, fertilization and embryo implantation. Hum Reprod Update. 2016;22(2):182–93. doi:10.1093/humupd/dmv055.

12.

Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116(2):281–97.CrossRefPubMed

13.

Lim LP, Lau NC, Garrett-Engele P, Grimson A, Schelter JM, Castle J, et al. Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature. 2005;433(7027):769–73. doi:10.1038/nature03315.CrossRefPubMed

14.

Li SC, Tang P, Lin WC. Intronic microRNA: discovery and biological implications. DNA Cell Biol. 2007;26(4):195–207. doi:10.1089/dna.2006.0558.CrossRefPubMed

15.

Krol J, Loedige I, Filipowicz W. The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet. 2010;11(9):597–610. doi:10.1038/nrg2843.PubMed

16.

Nagaraja AK, Andreu-Vieyra C, Franco HL, Ma L, Chen R, Han DY, et al. Deletion of Dicer in somatic cells of the female reproductive tract causes sterility. Mol Endocrinol. 2008;22(10):2336–52. doi:10.1210/me.2008-0142.CrossRefPubMedPubMedCentral

17.

Saetrom P, Snove Jr O, Rossi JJ. Epigenetics and microRNAs. Pediatr Res. 2007;61(5 Pt 2):17R–23R. doi:10.1203/pdr.0b013e318045760e.CrossRefPubMed

18.

Santonocito M, Vento M, Guglielmino MR, Battaglia R, Wahlgren J, Ragusa M, et al. Molecular characterization of exosomes and their microRNA cargo in human follicular fluid: bioinformatic analysis reveals that exosomal microRNAs control pathways involved in follicular maturation. Fertil Steril. 2014;102(6):1751–U590. doi:10.1016/j.fertnstert.2014.08.005.CrossRefPubMed

19.

da Silveira JC, de Andrade GM, Nogueira MF, Meirelles FV, Perecin F. Involvement of miRNAs and cell-secreted vesicles in mammalian ovarian antral follicle development. Reprod Sci. 2015;22(12):1474–83. doi:10.1177/1933719115574344.CrossRefPubMed

20.

Navakanitworakul R, Hung WT, Gunewardena S, Davis JS, Chotigeat W, Christenson LK. Characterization and small RNA content of extracellular vesicles in follicular fluid of developing bovine antral follicles. Sci Rep. 2016;6:25486. doi:10.1038/srep25486.CrossRefPubMedPubMedCentral

21.

da Silveira JC, Winger QA, Bouma GJ, Carnevale EM. Effects of age on follicular fluid exosomal microRNAs and granulosa cell transforming growth factor-beta signalling during follicle development in the mare. Reprod Fertil Dev. 2015;27(6):897–905. doi:10.1071/RD14452.CrossRefPubMed

22.

Sohel Md. Mahmodul H, Hoelker M, Noferesti SS, Salilew-Wondim D, Tholen E, Looft C et al. Exosomal and non-exosomal transport of extra-cellular microRNAs in follicular fluid: Implications for bovine oocyte developmental competence. PloS one. 2013;8(11).

23.

da Silveira JC, Veeramachaneni DNR, Winger QA, Carnevale EM, Bouma GJ. Cell-secreted vesicles in equine ovarian follicular fluid contain mirnas and proteins: A possible new form of cell communication within the ovarian follicle. Biology of Reproduction. 2012;86(3).

24.

Kim YJ, Ku SY, Kim YY, Liu HC, Chi SW, Kim SH, et al. MicroRNAs transfected into granulosa cells may regulate oocyte meiotic competence during in vitro maturation of mouse follicles. Hum Reprod. 2013;28(11):3050–61. doi:10.1093/humrep/det338.CrossRefPubMed

25.

Machtinger R, Bormann CL, Ginsburg ES, Racowsky C. Is the presence of a non-cleaved embryo on day 3 associated with poorer quality of the remaining embryos in the cohort? J Assist Reprod Genet. 2015. doi:10.1007/s10815-015-0455-9.PubMedPubMedCentral

26.

Alpha Scientists in Reproductive M, Embryology ESIGo. The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting. Hum Reprod. 2011;26(6):1270–83. doi:10.1093/humrep/der037.CrossRef

27.

Enderle D, Spiel A, Coticchia CM, Berghoff E, Mueller R, Schlumpberger M, et al. Characterization of RNA from exosomes and other extracellular vesicles isolated by a novel spin column-based method. PLoS One. 2015;10(8):e0136133. doi:10.1371/journal.pone.0136133.CrossRefPubMedPubMedCentral

28.

Racowsky C, Combelles CM, Nureddin A, Pan Y, Finn A, Miles L, et al. Day 3 and day 5 morphological predictors of embryo viability. Reprod Biomed Online. 2003;6(3):323–31.CrossRefPubMed

29.

Machtinger R, Racowsky C. Morphological systems of human embryo assessment and clinical evidence. Reprod Biomed Online. 2013;26(3):210–21. doi:10.1016/j.rbmo.2012.10.021.CrossRefPubMed

30.

Mestdagh P, Van Vlierberghe P, De Weer A, Muth D, Westermann F, Speleman F, et al. A novel and universal method for microRNA RT-qPCR data normalization. Genome Biol. 2009;10(6):R64. doi:10.1186/gb-2009-10-6-r64.CrossRefPubMedPubMedCentral

31.

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25(4):402–8. doi:10.1006/meth.2001.1262.CrossRefPubMed

32.

Agarwal V, Bell GW, Nam JW, Bartel DP. Predicting effective microRNA target sites in mammalian mRNAs. eLife. 2015;4. doi:10.7554/eLife.05005.

33.

Assou S, Al-edani T, Haouzi D, Philippe N, Lecellier CH, Piquemal D, et al. MicroRNAs: new candidates for the regulation of the human cumulus-oocyte complex. Hum Reprod. 2013;28(11):3038–49. doi:10.1093/humrep/det321.CrossRefPubMed

34.

Liu FJ, Shen XF. Comparative analysis of human reproductive proteomes identifies candidate proteins of sperm maturation. Mol Biol Rep. 2012;39(12):10257–63. doi:10.1007/s11033-012-1902-7.CrossRef

35.

Sang Q, Yao Z, Wang H, Feng R, Wang H, Zhao X, et al. Identification of MicroRNAs in human follicular fluid: characterization of MicroRNAs that govern steroidogenesis in vitro and are associated with polycystic ovary syndrome in vivo. J Clin Endocrinol Metab. 2013;98(7):3068–79.CrossRefPubMed

36.

Moreno JM, Nunez MJ, Quinonero A, Martinez S, de la Orden M, Simon C, et al. Follicular fluid and mural granulosa cells microRNA profiles vary in in vitro fertilization patients depending on their age and oocyte maturation stage. Fertil Steril. 2015;104(4):1037–46. doi:10.1016/j.fertnstert.2015.07.001. e1.CrossRefPubMed

37.

Velthut-Meikas A, Simm J, Tuuri T, Tapanainen JS, Metsis M, Salumets A. Research resource: small RNA-seq of human granulosa cells reveals miRNAs in FSHR and aromatase genes. Mol Endocrinol. 2013;27(7):1128–41. doi:10.1210/me.2013-1058.CrossRefPubMed

38.

Kishimoto T. Cell-cycle control during meiotic maturation. Curr Opin Cell Biol. 2003;15(6):654–63.CrossRefPubMed

39.

Stitzel ML, Seydoux G. Regulation of the oocyte-to-zygote transition. Science. 2007;316(5823):407–8. doi:10.1126/science.1138236.CrossRefPubMed

40.

Schier AF. The maternal-zygotic transition: death and birth of RNAs. Science. 2007;316(5823):406–7. doi:10.1126/science.1140693.CrossRefPubMed

41.

Giraldez AJ, Mishima Y, Rihel J, Grocock RJ, Van Dongen S, Inoue K, et al. Zebrafish MiR-430 promotes deadenylation and clearance of maternal mRNAs. Science. 2006;312(5770):75–9. doi:10.1126/science.1122689.CrossRefPubMed

42.

Whitaker M. Calcium at fertilization and in early development. Physiol Rev. 2006;86(1):25–88. doi:10.1152/physrev.00023.2005.CrossRefPubMedPubMedCentral

43.

Machaty Z. Signal transduction in mammalian oocytes during fertilization. Cell Tissue Res. 2016;363(1):169–83. doi:10.1007/s00441-015-2291-8.CrossRefPubMed

44.

Schatten H, Sun QY. New insights into the role of centrosomes in mammalian fertilization and implications for ART. Reproduction. 2011;142(6):793–801. doi:10.1530/REP-11-0261.CrossRefPubMed

45.

Mihaylova MM, Shaw RJ. The AMPK signalling pathway coordinates cell growth, autophagy and metabolism. Nat Cell Biol. 2011;13(9):1016–23. doi:10.1038/ncb2329.CrossRefPubMedPubMedCentral

46.

Cargnello M, Roux PP. Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. Microbiol Mol Biol Rev : MMBR. 2011;75(1):50–83. doi:10.1128/MMBR.00031-10.CrossRefPubMedPubMedCentral

47.

Odile ML, Heloise C, Julia M, Robert B, Patrick C. MAPK/ERK activity is required for the successful progression of mitosis in sea urchin embryos. Dev Biol. 2016. doi:10.1016/j.ydbio.2016.11.018.

48.

Feng R, Sang Q, Zhu Y, Fu W, Liu M, Xu Y, et al. MiRNA-320 in the human follicular fluid is associated with embryo quality in vivo and affects mouse embryonic development in vitro. Sci Rep. 2015;5:8689. doi:10.1038/srep08689.CrossRefPubMedPubMedCentral

49.

Kropp J, Khatib H. Characterization of microRNA in bovine in vitro culture media associated with embryo quality and development. J Dairy Sci. 2015;98(9):6552–63. doi:10.3168/jds.2015-9510.CrossRefPubMed

50.

Armstrong DA, Green BB, Seigne JD, Schned AR, Marsit CJ. MicroRNA molecular profiling from matched tumor and bio-fluids in bladder cancer. Mol Cancer. 2015;14:194. doi:10.1186/s12943-015-0466-2.CrossRefPubMedPubMedCentral

51.

Cha DJ, Franklin JL, Dou Y, Liu Q, Higginbotham JN, Demory Beckler M, et al. KRAS-dependent sorting of miRNA to exosomes. eLife. 2015;4:e07197. doi:10.7554/eLife.07197.CrossRefPubMedPubMedCentral

52.

Guzman N, Agarwal K, Asthagiri D, Yu L, Saji M, Ringel MD, et al. Breast cancer-specific miR signature unique to extracellular vesicles includes “microRNA-like” tRNA fragments. Mol Cancer Res : MCR. 2015;13(5):891–901. doi:10.1158/1541-7786.MCR-14-0533.CrossRefPubMedPubMedCentral

53.

Joshi GK, Deitz-McElyea S, Liyanage T, Lawrence K, Mali S, Sardar R, et al. Label-free nanoplasmonic-based short noncoding RNA sensing at attomolar concentrations allows for quantitative and highly specific assay of MicroRNA-10b in biological fluids and circulating exosomes. ACS Nano. 2015;9(11):11075–89. doi:10.1021/acsnano.5b04527.CrossRefPubMedPubMedCentral

54.

Melman YF, Shah R, Danielson K, Xiao J, Simonson B, Barth A, et al. Circulating MicroRNA-30d is associated with response to cardiac resynchronization therapy in heart failure and regulates cardiomyocyte apoptosis: a translational pilot study. Circulation. 2015;131(25):2202–16. doi:10.1161/CIRCULATIONAHA.114.013220.CrossRefPubMedPubMedCentral

Title: Extracellular microRNAs in follicular fluid and their potential association with oocyte fertilization and embryo quality: an exploratory study
Authors: Ronit Machtinger
Rodosthenis S. Rodosthenous
Michal Adir
Abdallah Mansour
Catherine Racowsky
Andrea A. Baccarelli
Russ Hauser
Publication date: 01-04-2017
Publisher: Springer US
Published in: Journal of Assisted Reproduction and Genetics / Issue 4/2017
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-017-0876-8

At a glance: The STEP trials

Springer Medicine

Extracellular microRNAs in follicular fluid and their potential association with oocyte fertilization and embryo quality: an exploratory study

Abstract

Purpose

Methods

Results

Conclusions

At a glance: The STEP trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 4/2017

Outcome of gestational surrogacy according to IVF protocol

Progesterone requires heat shock protein 90 (HSP90) in human sperm to regulate motility and acrosome reaction

Adiponectin and leptin gene polymorphisms in women with gestational diabetes mellitus

Redirecting reproductive immunology research toward pregnancy as a period of temporary immune tolerance

The impact of RABL2B gene (rs144944885) on human male infertility in patients with oligoasthenoteratozoospermia and immotile short tail sperm defects

Trendiness in human ARTs as technology transits from the macro to nano