The in vivo gene networks involved in coordinating human fetal ovarian development remain obscure. In this study, quantitative mass spectrometry was conducted on ovarian tissue collected at key stages during the first two trimesters of human gestational development, confirming the expression profiling data using immunofluorescence, as well as in vitro modeling with human oogonial stem cells (OSCs) and human embryonic stem cells (ESCs). A total of 3,837 proteins were identified in samples spanning developmental days 47–137. Bioinformatics clustering and Ingenuity Pathway Analysis identified DNA mismatch repair and base excision repair as major pathways upregulated during this time. In addition, MAEL and TEX11, two key meiosis-related proteins, were identified as highly expressed during the developmental window associated with fetal oogenesis. These findings were confirmed and extended using in vitro differentiation of OSCs into in vitro derived oocytes and of ESCs into primordial germ cell–like cells and oocyte–like cells, as models. In conclusion, the global protein expression profiling data generated by this study have provided novel insights into human fetal ovarian development in vivo and will serve as a valuable new resource for future studies of the signaling pathways used to orchestrate human oogenesis and folliculogenesis.
Get full access to this article
View all access options for this article.
References
1.
BakerTG and FranchiLL. (1967). The fine structure of oogonia and oocytes in human ovaries. J Cell Sci, 2:213–224.
2.
AndersonRA, FultonN, CowanG, CouttsS and SaundersPT. (2007). Conserved and divergent patterns of expression of DAZL, VASA and OCT4 in the germ cells of the human fetal ovary and testis. BMC Dev Biol, 7:136.
3.
GondosB. (1987). Comparative studies of normal and neoplastic ovarian germ cells: 2. Ultrastructure and pathogenesis of dysgerminoma. Int J Gynecol Pathol, 6:124–131.
4.
BendsenE, ByskovAG, AndersenCY and WestergaardLG. (2006). Number of germ cells and somatic cells in human fetal ovaries during the first weeks after sex differentiation. Hum Reprod, 21:30–35.
5.
MottaPM, MakabeS and NottolaSA. (1997). The ultrastructure of human reproduction. I. The natural history of the female germ cell: origin, migration and differentiation inside the developing ovary. Hum Reprod Update, 3:281–295.
6.
ForaboscoA and SforzaC. (2007). Establishment of ovarian reserve: a quantitative morphometric study of the developing human ovary. Fertil Steril, 88:675–683.
7.
AlbamonteMS, WillisMA, AlbamonteMI, JensenF, EspinosaMB and VitulloAD. (2008). The developing human ovary: immunohistochemical analysis of germ-cell-specific VASA protein, BCL-2/BAX expression balance and apoptosis. Hum Reprod, 23:1895–1901.
8.
PesceM, WangX, WolgemuthDJ and SchølerH. (1998). Differential expression of the Oct-4 transcription factor during mouse germ cell differentiation. Mech Dev, 71:89–98.
9.
StoopH, HoneckerF, CoolsM, de KrijgerR, BokemeyerC and LooijengaLH. (2005). Differentiation and development of human female germ cells during prenatal gonadogenesis: an immunohistochemical study. Hum Reprod, 20:1466–1476.
10.
HeJ, StewartK, KinnellHL, AndersonRA and ChildsAJ. (2013). A developmental stage-specific switch from DAZL to BOLL occurs during fetal oogenesis in humans, but not mice. PLoS One, 8:e73996.
11.
Le BouffantR, GuerquinMJ, DuquenneC, FrydmanN, CoffignyH, Rouiller-FabreV, FrydmanR, HabertR and LiveraG. (2010). Meiosis initiation in the human ovary requires intrinsic retinoic acid synthesis. Hum Reprod, 25:2579–2590.
12.
MamsenLS, ErnstEH, BorupR, LarsenA, OlesenRH, ErnstE, AndersonRA, KirstensenSG and AndersenCY. (2017). Temporal expression pattern of genes during the period of sex differentiation in human embryonic gonads. Sci Rep, 7:15961.
13.
HoumardB, SmallC, YangL, Naluai-CecchiniT, ChengE, HassoldT and GriswoldM. (2009). Global gene expression in the human fetal testis and ovary. Biol Reprod, 81:438–443.
14.
FowlerPA, FlanniganS, MathersA, GillandersK, LeaRG, WoodMJ, MaheshwariA, BhattacharyaS, Collie-DuguidES, et al. (2009). Gene expression analysis of human fetal ovarian primordial follicle formation. J Clin Endocrinol Metab, 94:1427–1435.
15.
SteuerwaldN, CohenJ, HerreraRJ, SandalinasM and BrennerCA. (2001). Association between spindle assembly checkpoint expression and maternal age in human oocytes. Mol Hum Reprod, 7:49–55.
16.
SteuerwaldNM, BermúdezMG, WellsD, MunnéS and CohenJ. (2007). Maternal age-related differential global expression profiles observed in human oocytes. Reprod Biomed Online, 14:700–708.
17.
GrøndahlML, AndersenCY, BogstadJ, NielsenFC, MeinertzH and BorupR. (2010). Gene expression profiles of single human mature oocytes in relation to age. Hum Reprod, 25:957–968.
18.
WoodJR, DumesicDA, AbbottDH and StraussJF3rd. (2007). Molecular abnormalities in oocytes from women with polycystic ovary syndrome revealed by microarray analysis. J Clin Endocrinol Metab, 92:705–713.
19.
MarkholtS, GrøndahlML, ErnstEH, AnderseCY, ErnstE and Lykke-HartmannK. (2012). Global gene analysis of oocytes from early stages in human folliculogenesis shows high expression of novel genes in reproduction. Mol Hum Reprod, 18:96–110.
20.
JonesGM, CramDS, SongB, MagliMC, GianaroliL, Lacham-KaplanO, FindlayJK, JenkinG and TrounsonAO. (2008). Gene expression profiling of human oocytes following in vivo or in vitro maturation. Hum Reprod, 23:1138–1144.
21.
HikabeO, HamazakiN, NagamatsuG, ObataY, HiraoY, HamadaN, ShimamotoS, ImamuraT, NakashimaK, SaitouM and HayashiK. (2016). Reconstitution in vitro of the entire cycle of the mouse female germ line. Nature, 539:299–303.
22.
IrieN, WeinbergerL, TangWWC, KobayashiT, ViukovS, ManorYS, DietmannS, HannaJH and SuraniMA. (2015). Sox17 is a critical specifier of human primordial germ cell fate. Cell, 160:253–268.
23.
SasakiK, YokobayashiS, NakamuraT, OkamotoI, YabutaY, KurimotoK, OhtaH, MoritokiY, IwataniC, et al. (2015). Robust in vitro induction of human germ cell fate from pluripotent stem cells. Cell Stem Cell, 17:178–194.
24.
JungD, XiongJ, YeM, QinX, LiL, ChengS, LuoM, PengJ, DongJ, et al. (2017). In vitro differentiation of human embryonic stem cells into ovarian follicle-like cells. Nat Commun, 8:15680.
25.
WhiteYAR, WoodsDC, TakaiY, IshiharaO, SekiH and TillyJL. (2012). Oocyte formation by mitotically active germ cells purified from ovaries of reproductive-age women. Nat Med, 18:413–421.
26.
WoodsDC and TillyJL. (2013). Isolation, characterization and propogation of mitotically active germ cells from adult mouse and human ovaries. Nat Protoc, 8:966–988.
27.
WoodsDC and TillyJL. (2015). Autologous germline mitochondrial energy transfer (AUGMENT) in human assisted reproduction. Semin Reprod Med, 33:410–421.
28.
SchwanhausserB, BusseD, LiN, DittmarG, SchuchhardtJ, WolfJ, ChenW and SelbachM. (2011). Global quantification of mammalian gene expression control. Nature, 473:337–342.
29.
TamayoP, SlonimD, MesirovJ, ZhuQ, KitareewanS, DmitrovskyE, LanderES and GolubTR. (1999). Interpreting patterns of gene expression with self-organizing maps: methods and application to hematopoietic differentiation. Proc Natl Acad Sci U S A, 96:2907–2912.
HeberleH, Vaz MeirellesG, da SilvaFR, TellesGP and MinghimR. (2015). InteractiVenn: a web-based tool for the analysis of sets through Venn diagrams. BMC Bioinformatics, 16:169.
32.
VizcainoJA, CoteRG, CsordasA, DianesJA, FabregatA, FosterJM, GrissJ, ApliE, BirimM, et al. (2013). The proteomics identifications (PRIDE) database and associated tools: status in 2013. Nucleic Acids Res, 41:D1063–D1069.
33.
KuriloLF. (1981). Oogenesis in antenatal development in man. Hum Genet, 57:86–92.
34.
KonishiI, FujiiS, OkamuraH, ParmleyT and MoriT. (1986). Development of interstitial cells and ovigerous cords in the human fetal ovary: an ultrastructural study. J Anat, 148:121–135.
35.
SatoK, NishidaKM, ShibuyaA, SiomiMC and SiomiH. (2011). Maelstrom coordinates microtubule organization during Drosophila oogenesis through interaction with components of the MTOC. Genes Dev, 25:2361–2373.
36.
AdelmanCA and PetriniJH. (2008). ZIP4H (TEX11) deficiency in the mouse impairs meiotic double strand break repair and the regulation of crossing over. PLoS Genet, 4:e1000042.
37.
DingX, LiuG, XuB, WuC, HuiN, NiX, WangJ, DuM, TengX and WuJ. (2016). Human GV oocytes generated by mitotically active germ cells obtained from follicular aspirates. Sci Rep, 6:28218.
38.
ClarkAT, BodnarMS, FoxM, RodriquezRT, AbeytaMJ, FirpoMT and PeraRA. (2004). Spontaneous differentiation of germ cells from human embryonic stem cells in vitro. Hum Mol Genet, 13:727–739.
39.
ChenHF, KuoHC, ChienCL, ShunCT, YaoYL, IpPL, ChuangCY, WangCC, YangYS and HoHN. (2007). Derivation, characterization and differentiation of human embryonic stem cells: comparing serum-containing versus serum-free media and evidence of germ cell differentiation. Hum Reprod, 22:567–577.
40.
TilgnerK, AtkinsonSP, GolebiewskaA, StojkovicM, LakoM and ArmstrongL. (2008). Isolation of primordial germ cells from differentiating human embryonic stem cells. Stem Cells, 26:3075–3085.
41.
GuW, TekurS, ReinboldR, EppigJJ, ChoiYC, ZhengeJZ, MurrayMT and HechtNB. (1998). Mammalian male and female germ cells express a germ cell-specific Y-Box protein, MSY2. Biol Reprod, 59:1266–1274.
42.
YangJ, MedvedexS, YuJ, TangLC, AgnoJE, MatzukMM, SchultzRM and HechtNB. (2005). Absence of the DNA-/RNA-binding protein MSY2 results in male and female infertility. Proc Natl Acad Sci U S A, 102:5755–5760.
43.
PageSL and HawleyRS. (2004). The genetics and molecular biology of the synaptonemal complex. Annu Rev Cell Dev Biol, 20:525–558.
44.
YuanL, LiuJG, HojaMR, WilbertzJ, NordqvistK and HoogC. (2002). Female germ cell aneuploidy and embryo death in mice lacking the meiosis-specific protein SCP3. Science, 296:1115–1118.
45.
SmithRW, AndersonRC, SmithJW, BrookM, RichardsonWA and GrayNK. (2011). DAZAP1, an RNA-binding protein required for development and spermatogenesis, can regulate mRNA translation. RNA, 17:1282–1295.
46.
EdelmannW, CohenPE, KneitzB, WinandN, LiaM, HeyerJ, KolodnerR, PollardJW and KucherlapatiR. (1999). Mammalian MutS homologue 5 is required for chromosome pairing in meiosis. Nat Genet, 21:123–127.
47.
de VriesSS, BaartEB, DekkerM, SiezenA, de RooijDG, de BoerP and te RieleH. (1999). Mouse MutS-like protein Msh5 is required for proper chromosome synapsis in male and female meiosis. Genes Dev, 13:523–531.
48.
KneitzB, CohenPE, AvdievichE, ZhuL, KaneMF, HouHJr, KolodnerRD, KucherlapatiR, PollardJW and EdelmannW. (2000). MutS homolog 4 localization to meiotic chromosomes is required for chromosome pairing during meiosis in male and female mice. Genes Dev, 14:1085–1097.
49.
JaroudiS, KakourouG, CawoodS, DoshiA, RanieriDM, SerhalP, HarperJC and SenGuptaSB. (2009). Expression profiling of DNA repair gnes in human oocytes and blastocysts using microarrays. Hum Reprod, 24:2649–2655.
50.
ChildsAJ, CowanG, KinnellHL, AndersonRA and SaundersPT. (2011). Retinoic acid signaling and the control of meiotic entry in the human fetal gonad. PLoS One, 6:e20249.
51.
Stoop-MyerC and AmonA. (1999). Meiosis: Rec8 is the reason for cohesion. Nat Cell Biol, 1:E125–E127.
52.
Garcia-CruzR, BrieñoMA, RoigI, GrossmannM, VelillaE, PujolA, CaberoL, PessarrodonaA, BarberoJL and Garcia CaldésM. (2010). Dynamics of cohesin proteins REC8, STAG3, SMC1 beta and SMC3 are consistent with a role in sister chromatid cohesion during meiosis in human oocytes. Hum Reprod, 25:2316–2327.
53.
HopkinsJ, HwangG, JacobJ, SappN, BedigianR, OkaK, OverbeekP, MurrayS and JordanPW. (2014). Meiosis-specific cohesin component, Stag3 is essential for maintaining centromere chromatid cohesion, and required for DNA repair and synapsis between homologous chromosomes. PLoS Genet, 10:e1004413.
54.
SyrjanenJL, PellegriniL and DaviesOR. (2014). A molecular model for the role of SYCP3 in meiotic chromosome organisation. Elife, 3:E02963.
55.
SaxeJP, ChenM, ZhaoH and LinH. (2013). Tdrkh is essential for spermatogenesis and participates in primary piRNA biogenesis in the germline. EMBO J, 32:1869–1885.
56.
SoperSF, van Der HeijdenGW, HardimanTC, GoodheartM, MartinSL, de BoerP and BortvinA. (2008). Mouse maelstrom, a component of nuage, is essential for spermatogenesis and transposon repression in meiosis. Dev Cell, 15:285–297.
57.
MalkiS, van der HeijdenGW, O'DonnellKA, MartinSL and BortvinA. (2014). A role for retrotransposon LINE-1 in fetal oocyte attrition in mice. Dev Cell, 29:521–533.
58.
XiaoL, WangY, ZhouY, SunY, SunW, WangL, ZhouC, ZhouJ and ZhangJ. (2010). Identification of a novel human cancer/testis gene MAEL that is regulated by DNA methylation. Mol Biol Rep, 37:2355–2360.
59.
LiL, DongJ, YanL, YongJ, LiuX, HuY, FanX, WuW, GuoH, et al. (2017). Single-cell RNA-Seq analysis maps development of human germline cells and gonadal niche interactions. Cell Stem Cell, 20:1–16.
60.
RosarioR, SmithRWP, AdamsIR and AndersonRA. (2017). RNA immunoprecipitation identifies novel targets of DAZL in human foetal ovary. Mol Hum Reprod, 23:177–186.
61.
YatsenkoAN, GeorgiadisAP, RöpkeA, BermanAJ, JaffeT, OlszewskaM, WesternströerB, SanfilippoJ, KurpiszM, et al. (2015). X-linked TEX11 mutations, meiotic arrest, and azoospermia in infertile men. N Engl J Med, 372:2097–2107.
62.
RichardsJS, RussellDL, OchsnerS and EspeyLL. (2002). Ovulation: new dimensions and new regulators of the inflammatory-like response. Annu Rev Physiol, 64:69–92.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
