Cell-cell communication within the follicle involves many signaling molecules, and this process may be mediated by secretion and uptake of exosomes that contain several bioactive molecules including extra-cellular miRNAs. Follicular fluid and cells from individual follicles of cattle were grouped based on Brilliant Cresyl Blue (BCB) staining of the corresponding oocytes. Both Exoquick precipitation and differential ultracentrifugation were used to separate the exosome and non-exosomal fraction of follicular fluid. Following miRNA isolation from both fractions, the human miRCURY LNA? Universal RT miRNA PCR array system was used to profile miRNA expression. This analysis found that miRNAs were present in both exosomal and non-exosomal fraction of bovine follicular fluid. We found 25 miRNAs differentially expressed (16 up and 9 down) in exosomes and 30 miRNAs differentially expressed (21 up and 9 down) in non-exosomal fraction of follicular fluid in comparison of BCB- versus BCB+ oocyte groups. Expression of selected miRNAs was detected in theca, granulosa and cumulus oocyte complex. To further explore the potential roles of these follicular fluid derived extra-cellular miRNAs, the potential target genes were predicted, and functional annotation and pathway analysis revealed most of these pathways are known regulators of follicular development and oocyte growth. In order to validate exosome mediated cell-cell communication within follicular microenvironment, we demonstrated uptake of exosomes and resulting increase of endogenous miRNA level and subsequent alteration of mRNA levels in follicular cells in vitro. This study demonstrates for the first time, the presence of exosome or non-exosome mediated transfer of miRNA in the bovine follicular fluid, and oocyte growth dependent variation in extra-cellular miRNA signatures in the follicular environment.
References
[1]
Eppig JJ (2001) Oocyte control of ovarian follicular development and function in mammals. Reproduction 122: 829–838.
[2]
Harwood BN, Cross SK, Radford EE, Haac BE, De Vries WN (2008) Members of the WNT signaling pathways are widely expressed in mouse ovaries, oocytes, and cleavage stage embryos. Dev Dyn 237: 1099–1111.
[3]
Luo S, Kleemann GA, Ashraf JM, Shaw WM, Murphy CT (2010) TGF-beta and insulin signaling regulate reproductive aging via oocyte and germline quality maintenance. Cell 143: 299–312.
[4]
Su YQ, Wu X, O'Brien MJ, Pendola FL, Denegre JN, et al. (2004) Synergistic roles of BMP15 and GDF9 in the development and function of the oocyte-cumulus cell complex in mice: genetic evidence for an oocyte-granulosa cell regulatory loop. Dev Biol 276: 64–73.
[5]
Patsoula E, Loutradis D, Drakakis P, Kallianidis K, Bletsa R, et al. (2001) Expression of mRNA for the LH and FSH receptors in mouse oocytes and preimplantation embryos. Reproduction 121: 455–461.
[6]
Krisher RL (2004) The effect of oocyte quality on development. J Anim Sci 82 Suppl: E14–23
[7]
Sirard MA, Richard F, Blondin P, Robert C (2006) Contribution of the oocyte to embryo quality. Theriogenology 65: 126–136.
[8]
Silva DS, Rodriguez P, Galuppo A, Arruda NS, Rodrigues JL (2011) Selection of bovine oocytes by brilliant cresyl blue staining: effect on meiosis progression, organelle distribution and embryo development. Zygote: 1–6.
[9]
Pujol M, Lopez-Bejar M, Paramio MT (2004) Developmental competence of heifer oocytes selected using the brilliant cresyl blue (BCB) test. Theriogenology 61: 735–744.
[10]
Catala MG, Izquierdo D, Uzbekova S, Morato R, Roura M, et al. (2011) Brilliant Cresyl Blue stain selects largest oocytes with highest mitochondrial activity, maturation-promoting factor activity and embryo developmental competence in prepubertal sheep. Reproduction 142: 517–527.
[11]
Armstrong DG, Webb R (1997) Ovarian follicular dominance: the role of intraovarian growth factors and novel proteins. Rev Reprod 2: 139–146.
[12]
Gosden RG, Hunter RH, Telfer E, Torrance C, Brown N (1988) Physiological factors underlying the formation of ovarian follicular fluid. J Reprod Fertil 82: 813–825.
[13]
Revelli A, Delle Piane L, Casano S, Molinari E, Massobrio M, et al. (2009) Follicular fluid content and oocyte quality: from single biochemical markers to metabolomics. Reprod Biol Endocrinol 7: 40.
[14]
McNatty KP, Baird DT (1978) Relationship between follicle-stimulating hormone, androstenedione and oestradiol in human follicular fluid. J Endocrinol 76: 527–531.
[15]
Espey LL (1994) Current status of the hypothesis that mammalian ovulation is comparable to an inflammatory reaction. Biol Reprod 50: 233–238.
[16]
Mittelbrunn M, Gutierrez-Vazquez C, Villarroya-Beltri C, Gonzalez S, Sanchez-Cabo F, et al. (2011) Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells. Nat Commun 2: 282.
[17]
Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, et al. (2007) Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 9: 654–659.
[18]
Taylor DD, Gercel-Taylor C (2008) MicroRNA signatures of tumor-derived exosomes as diagnostic biomarkers of ovarian cancer. Gynecol Oncol 110: 13–21.
[19]
Pegtel DM, Cosmopoulos K, Thorley-Lawson DA, van Eijndhoven MA, Hopmans ES, et al. (2010) Functional delivery of viral miRNAs via exosomes. Proc Natl Acad Sci U S A 107: 6328–6333.
[20]
da Silveira JC, Veeramachaneni DN, Winger QA, Carnevale EM, Bouma GJ (2011) Cell-Secreted Vesicles in Equine Ovarian Follicular Fluid Contain miRNAs and Proteins: A Possible New Form of Cell Communication Within the Ovarian Follicle. Biol Reprod 86: 71.
[21]
Hossain MM, Sohel MM, Schellander K, Tesfaye D (2012) Characterization and importance of microRNAs in mammalian gonadal functions. Cell Tissue Res 349: 679–690.
[22]
Baley J, Li J (2012) MicroRNAs and ovarian function. J Ovarian Res 5: 8.
[23]
Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, et al. (2008) Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci U S A 105: 10513–10518.
[24]
Park NJ, Zhou H, Elashoff D, Henson BS, Kastratovic DA, et al. (2009) Salivary microRNA: discovery, characterization, and clinical utility for oral cancer detection. Clin Cancer Res 15: 5473–5477.
[25]
Weber JA, Baxter DH, Zhang S, Huang DY, Huang KH, et al. (2010) The microRNA spectrum in 12 body fluids. Clin Chem 56: 1733–1741.
[26]
Simons M, Raposo G (2009) Exosomes—vesicular carriers for intercellular communication. Curr Opin Cell Biol 21: 575–581.
[27]
Arroyo JD, Chevillet JR, Kroh EM, Ruf IK, Pritchard CC, et al. (2010) Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma. Proc Natl Acad Sci U S A 108: 5003–5008.
[28]
Cortez MA, Welsh JW, Calin GA (2012) Circulating microRNAs as noninvasive biomarkers in breast cancer. Recent Results Cancer Res 195: 151–161.
[29]
Ghanem N, Holker M, Rings F, Jennen D, Tholen E, et al. (2007) Alterations in transcript abundance of bovine oocytes recovered at growth and dominance phases of the first follicular wave. BMC Dev Biol 7: 90.
[30]
Spanel-Borowski K, Ricken AM, Saxer M, Huber PR (1994) Long-term co-culture of bovine granulosa cells with microvascular endothelial cells: effect on cell growth and cell death. Mol Cell Endocrinol 104: 11–19.
[31]
Wang WX, Wilfred BR, Hu Y, Stromberg AJ, Nelson PT (2009) Anti-Argonaute RIP-Chip shows that miRNA transfections alter global patterns of mRNA recruitment to microribonucleoprotein complexes. RNA 16: 394–404.
[32]
Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP, Burge CB (2003) Prediction of mammalian microRNA targets. Cell 115: 787–798.
[33]
Brennecke J, Stark A, Russell RB, Cohen SM (2005) Principles of microRNA-target recognition. PLoS Biol 3: e85.
[34]
Tesfaye D, Worku D, Rings F, Phatsara C, Tholen E, et al. (2009) Identification and expression profiling of microRNAs during bovine oocyte maturation using heterologous approach. Mol Reprod Dev 76: 665–677.
[35]
Gad A, Besenfelder U, Rings F, Ghanem N, Salilew-Wondim D, et al. (2011) Effect of reproductive tract environment following controlled ovarian hyperstimulation treatment on embryo development and global transcriptome profile of blastocysts: implications for animal breeding and human assisted reproduction. Hum Reprod 26: 1693–1707.
[36]
Thery C (2011) Exosomes: secreted vesicles and intercellular communications. F1000 Biol Rep 3: 15.
[37]
Cui W, Ma J, Wang Y, Biswal S (2011) Plasma miRNA as biomarkers for assessment of total-body radiation exposure dosimetry. PLoS One 6: e22988.
[38]
Yamaura Y, Nakajima M, Takagi S, Fukami T, Tsuneyama K, et al. (2012) Plasma microRNA profiles in rat models of hepatocellular injury, cholestasis, and steatosis. PLoS One 7: e30250.
[39]
Hunter MP, Ismail N, Zhang X, Aguda BD, Lee EJ, et al. (2008) Detection of microRNA expression in human peripheral blood microvesicles. PLoS One 3: e3694.
[40]
Gilad S, Meiri E, Yogev Y, Benjamin S, Lebanony D, et al. (2008) Serum microRNAs are promising novel biomarkers. PLoS One 3: e3148.
[41]
Schrauder MG, Strick R, Schulz-Wendtland R, Strissel PL, Kahmann L, et al. (2012) Circulating micro-RNAs as potential blood-based markers for early stage breast cancer detection. PLoS One 7: e29770.
[42]
Logozzi M, De Milito A, Lugini L, Borghi M, Calabro L, et al. (2009) High levels of exosomes expressing CD63 and caveolin-1 in plasma of melanoma patients. PLoS One 4: e5219.
[43]
Gallo A, Tandon M, Alevizos I, Illei GG (2012) The majority of microRNAs detectable in serum and saliva is concentrated in exosomes. PLoS One 7: e30679.
[44]
Torri AF, Hajduk SL (1988) Posttranscriptional regulation of cytochrome c expression during the developmental cycle of Trypanosoma brucei. Mol Cell Biol 8: 4625–4633.
[45]
Vickers KC, Palmisano BT, Shoucri BM, Shamburek RD, Remaley AT (2011) MicroRNAs are transported in plasma and delivered to recipient cells by high-density lipoproteins. Nat Cell Biol 13: 423–433.
[46]
Bentwich I, Avniel A, Karov Y, Aharonov R, Gilad S, et al. (2005) Identification of hundreds of conserved and nonconserved human microRNAs. Nat Genet 37: 766–770.
[47]
Fair T, Hyttel P, Greve T (1995) Bovine oocyte diameter in relation to maturational competence and transcriptional activity. Mol Reprod Dev 42: 437–442.
[48]
Huo LJ, Fan HY, Zhong ZS, Chen DY, Schatten H, et al. (2004) Ubiquitin-proteasome pathway modulates mouse oocyte meiotic maturation and fertilization via regulation of MAPK cascade and cyclin B1 degradation. Mech Dev 121: 1275–1287.
[49]
Suzumori N, Burns KH, Yan W, Matzuk MM (2003) RFPL4 interacts with oocyte proteins of the ubiquitin-proteasome degradation pathway. Proc Natl Acad Sci U S A 100: 550–555.
[50]
Ciechanover A, Schwartz AL (1994) The ubiquitin-mediated proteolytic pathway: mechanisms of recognition of the proteolytic substrate and involvement in the degradation of native cellular proteins. FASEB J 8: 182–191.
[51]
Dissen GA, Garcia-Rudaz C, Ojeda SR (2009) Role of neurotrophic factors in early ovarian development. Semin Reprod Med 27: 24–31.
[52]
Zhang M, Ouyang H, Xia G (2009) The signal pathway of gonadotrophins-induced mammalian oocyte meiotic resumption. Mol Hum Reprod 15: 399–409.
[53]
Boyer A, Goff AK, Boerboom D (2010) WNT signaling in ovarian follicle biology and tumorigenesis. Trends Endocrinol Metab 21: 25–32.
[54]
Zheng P, Vassena R, Latham K (2006) Expression and downregulation of WNT signaling pathway genes in rhesus monkey oocytes and embryos. Mol Reprod Dev 73: 667–677.
[55]
Pritchard CC, Kroh E, Wood B, Arroyo JD, Dougherty KJ, et al. (2011) Blood cell origin of circulating microRNAs: a cautionary note for cancer biomarker studies. Cancer Prev Res (Phila) 5: 492–497.
[56]
Zhou Q, Li M, Wang X, Li Q, Wang T, et al. (2012) Immune-related microRNAs are abundant in breast milk exosomes. Int J Biol Sci 8: 118–123.
[57]
Fitzner D, Schnaars M, van Rossum D, Krishnamoorthy G, Dibaj P, et al. (2011) Selective transfer of exosomes from oligodendrocytes to microglia by macropinocytosis. J Cell Sci 124: 447–458.
[58]
Tian T, Wang Y, Wang H, Zhu Z, Xiao Z (2010) Visualizing of the cellular uptake and intracellular trafficking of exosomes by live-cell microscopy. J Cell Biochem 111: 488–496.
[59]
Penna E, Orso F, Cimino D, Tenaglia E, Lembo A, et al. (2011) microRNA-214 contributes to melanoma tumour progression through suppression of TFAP2C. EMBO J 30: 1990–2007.
[60]
Mongan M, Wang J, Liu H, Fan Y, Jin C, et al. (2011) Loss of MAP3K1 enhances proliferation and apoptosis during retinal development. Development 138: 4001–4012.
[61]
Sutherland JM, Keightley RA, Nixon B, Roman SD, Robker RL, et al. (2011) Suppressor of cytokine signaling 4 (SOCS4): moderator of ovarian primordial follicle activation. J Cell Physiol 227: 1188–1198.
[62]
Forde N, Duffy GB, McGettigan PA, Browne JA, Mehta JP, et al. (2012) Evidence for an early endometrial response to pregnancy in cattle: both dependent upon and independent of interferon tau. Physiol Genomics 44: 799–810.
[63]
Ding F, Li HH, Li J, Myers RM, Francke U (2010) Neonatal maternal deprivation response and developmental changes in gene expression revealed by hypothalamic gene expression profiling in mice. PLoS One 5: e9402.
[64]
Yokoo M, Miyahayashi Y, Naganuma T, Kimura N, Sasada H, et al. (2002) Identification of hyaluronic acid-binding proteins and their expressions in porcine cumulus-oocyte complexes during in vitro maturation. Biol Reprod 67: 1165–1171.
[65]
Araujo VR, Silva GM, Duarte AB, Magalhaes DM, Almeida AP, et al. (2011) Vascular endothelial growth factor-A(165) (VEGF-A(165)) stimulates the in vitro development and oocyte competence of goat preantral follicles. Cell Tissue Res 346: 273–281.