Abstract
Obesity impairs reproductive capacity, and the link between imprinting disorders and obesity has been discussed in many studies. Recent studies indicate that a high-fat diet may cause epigenetic changes in maternal and paternal genes, which may be transmitted to offspring and negatively affect their development. On this basis, our study aims to reveal the changes in DNA methylation and DNA methyltransferase enzymes in the ovaries and testes of C57BL/6 mice fed a high-fat diet and created a model of obesity, by comparing them with the control group. For this purpose, we demonstrated the presence and quantitative differences of DNA methyltransferase 1 and DNA methyltransferase 3a enzymes as well as global DNA methylation in ovaries and testis of C57BL/6 mice fed a high-fat diet by using immunohistochemistry and western blot methods. We found that a high-fat diet induces the levels of Dnmt1 and Dnmt3a proteins (p < 0.05). We observed increased global DNA methylation in testes but, interestingly, decreased global DNA methylation in ovaries. We think that our outcomes have significant value to demonstrate the effects of obesity on ovarian follicle development and testicular spermatogenesis and may bring a new perspective to obesity-induced infertility treatments. Additionally, to the best of our knowledge, this is the first study to document dynamic alteration of Dnmt1 and Dnmt3a as well as global DNA methylation patterns during follicle development in healthy mouse ovaries.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Code availability
Not applicable.
Data availability
Data and material available upon request.
References
Atzmon Y, Shoshan-Karchovsky E, Michaeli M, Aslih N, Shrem G, Ellenbogen A, Shalom-Paz E (2017) Obesity results with smaller oocyte in in vitro fertilization/intracytoplasmic sperm injection cycles—a prospective study. J Assist Reprod Genet 34(9):1145–1151. https://doi.org/10.1007/s10815-017-0975-6
Barbagallo F, Condorelli RA, Mongioi LM, Cannarella R, Cimino L, Magagnini MC, Crafa A, La Vignera S, Calogero AE (2021) Molecular mechanisms underlying the relationship between obesity and male infertility. Metabolites 11(12):840. https://doi.org/10.3390/metabo11120840
Campion J, Milagro FI, Martinez JA (2009) Individuality and epigenetics in obesity. Obes Rev 10(4):383–392. https://doi.org/10.1111/j.1467-789X.2009.00595.x
Catalano PM, Shankar K (2017) Obesity and pregnancy: mechanisms of short term and long term adverse consequences for mother and child. BMJ 356:j1. https://doi.org/10.1136/bmj.j1
Cheong Y, Sadek KH, Bruce KD, Macklon N, Cagampang FR (2014) Diet-induced maternal obesity alters ovarian morphology and gene expression in the adult mouse offspring. Fertil Steril 102(3):899–907. https://doi.org/10.1016/j.fertnstert.2014.06.015
Collaboration NCDRF (2016) Trends in adult body-mass index in 200 countries from 1975 to 2014: a pooled analysis of 1698 population-based measurement studies with 19.2 million participants. Lancet 387(10026):1377–1396. https://doi.org/10.1016/S0140-6736(16)30054-X
Dag ZO, Dilbaz B (2015) Impact of obesity on infertility in women. J Turk Ger Gynecol Assoc 16(2):111–117. https://doi.org/10.5152/jtgga.2015.15232
Davidson LM, Millar K, Jones C, Fatum M, Coward K (2015) Deleterious effects of obesity upon the hormonal and molecular mechanisms controlling spermatogenesis and male fertility. Hum Fertil (Camb) 18(3):184–193. https://doi.org/10.3109/14647273.2015.1070438
Dean W, Santos F, Reik W (2003) Epigenetic reprogramming in early mammalian development and following somatic nuclear transfer. Semin Cell Dev Biol 14(1):93–100. https://doi.org/10.1016/s1084-9521(02)00141-6
Deshpande SSS, Nemani H, Balasinor NH (2021) High fat diet-induced- and genetically inherited- obesity differential alters DNA demethylation pathways in the germline of adult male rats. Reprod Biol 21(3):100532. https://doi.org/10.1016/j.repbio.2021.100532
Fullston T, Ohlsson Teague EM, Palmer NO, DeBlasio MJ, Mitchell M, Corbett M, Print CG, Owens JA, Lane M (2013) Paternal obesity initiates metabolic disturbances in two generations of mice with incomplete penetrance to the F2 generation and alters the transcriptional profile of testis and sperm microRNA content. FASEB J 27(10):4226–4243. https://doi.org/10.1096/fj.12-224048
Fullston T, McPherson NO, Owens JA, Kang WX, Sandeman LY, Lane M (2015) Paternal obesity induces metabolic and sperm disturbances in male offspring that are exacerbated by their exposure to an obesogenic diet. Physiol Rep 3(3):e12336. https://doi.org/10.14814/phy2.12336
Huang Q, Ma C, Chen L, Luo D, Chen R, Liang F (2018) Mechanistic insights into the interaction between transcription factors and epigenetic modifications and the contribution to the development of obesity. Front Endocrinol (Lausanne) 9:370. https://doi.org/10.3389/fendo.2018.00370
Kageyama S, Liu H, Kaneko N, Ooga M, Nagata M, Aoki F (2007) Alterations in epigenetic modifications during oocyte growth in mice. Reproduction 133(1):85–94. https://doi.org/10.1530/REP-06-0025
Kasum M, Oreskovic S, Cehic E, Lila A, Ejubovic E, Soldo D (2018) The role of female obesity on in vitro fertilization outcomes. Gynecol Endocrinol 34(3):184–188. https://doi.org/10.1080/09513590.2017.1391209
Kroes M, Osei-Assibey G, Baker-Searle R, Huang J (2016) Impact of weight change on quality of life in adults with overweight/obesity in the United States: a systematic review. Curr Med Res Opin 32(3):485–508. https://doi.org/10.1185/03007995.2015.1128403
Kurihara Y, Kawamura Y, Uchijima Y, Amamo T, Kobayashi H, Asano T, Kurihara H (2008) Maintenance of genomic methylation patterns during preimplantation development requires the somatic form of DNA methyltransferase 1. Dev Biol 313(1):335–346. https://doi.org/10.1016/j.ydbio.2007.10.033
Lainez NM, Coss D (2019) Obesity, neuroinflammation, and reproductive function. Endocrinology 160(11):2719–2736. https://doi.org/10.1210/en.2019-00487
Liu Y, Ding Z (2017) Obesity, a serious etiologic factor for male subfertility in modern society. Reproduction 154(4):R123–R131. https://doi.org/10.1530/REP-17-0161
Lucifero D, Mann MR, Bartolomei MS, Trasler JM (2004) Gene-specific timing and epigenetic memory in oocyte imprinting. Hum Mol Genet 13(8):839–849. https://doi.org/10.1093/hmg/ddh104
Marques CJ, Joao Pinho M, Carvalho F, Bieche I, Barros A, Sousa M (2011) DNA methylation imprinting marks and DNA methyltransferase expression in human spermatogenic cell stages. Epigenetics 6(11):1354–1361. https://doi.org/10.4161/epi.6.11.17993
Maugeri A (2020) The effects of dietary interventions on DNA methylation: implications for obesity management. Int J Mol Sci 21(22):8670. https://doi.org/10.3390/ijms21228670
Mitchell M, Armstrong DT, Robker RL, Norman RJ (2005) Adipokines: implications for female fertility and obesity. Reproduction 130(5):583–597. https://doi.org/10.1530/rep.1.00521
Okano M, Xie S, Li E (1998) Cloning and characterization of a family of novel mammalian DNA (cytosine-5) methyltransferases. Nat Genet 19(3):219–220. https://doi.org/10.1038/890
Okano M, Bell DW, Haber DA, Li E (1999) DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell 99(3):247–257. https://doi.org/10.1016/s0092-8674(00)81656-6
Omisanjo OA, Biermann K, Hartmann S, Heukamp LC, Sonnack V, Hild A, Brehm R, Bergmann M, Weidner W, Steger K (2007) DNMT1 and HDAC1 gene expression in impaired spermatogenesis and testicular cancer. Histochem Cell Biol 127(2):175–181. https://doi.org/10.1007/s00418-006-0234-x
Ooi SL, Henikoff S (2007) Germline histone dynamics and epigenetics. Curr Opin Cell Biol 19(3):257–265. https://doi.org/10.1016/j.ceb.2007.04.015
Ozanne SE (2015) Epigenetic signatures of obesity. N Engl J Med 372(10):973–974. https://doi.org/10.1056/NEJMcibr1414707
Pan Z, Zhang J, Li Q, Li Y, Shi F, Xie Z, Liu H (2012) Current advances in epigenetic modification and alteration during mammalian ovarian folliculogenesis. J Genet Genom 39(3):111–123. https://doi.org/10.1016/j.jgg.2012.02.004
Potabattula R, Dittrich M, Schorsch M, Hahn T, Haaf T, El Hajj N (2019) Male obesity effects on sperm and next-generation cord blood DNA methylation. PLoS ONE 14(6):e0218615. https://doi.org/10.1371/journal.pone.0218615
Practice Committee of the American Society for Reproductive Medicine. Electronic address aao, Practice Committee of the American Society for Reproductive M (2021) Obesity and reproduction: a committee opinion. Fertil Steril 116(5):1266–1285. https://doi.org/10.1016/j.fertnstert.2021.08.018
Rao KR, Lal N, Giridharan NV (2014) Genetic & epigenetic approach to human obesity. Indian J Med Res 140(5):589–603
Rhee JS, Saben JL, Mayer AL, Schulte MB, Asghar Z, Stephens C, Chi MM, Moley KH (2016) Diet-induced obesity impairs endometrial stromal cell decidualization: a potential role for impaired autophagy. Hum Reprod 31(6):1315–1326. https://doi.org/10.1093/humrep/dew048
Robertson KD (2005) DNA methylation and human disease. Nat Rev Genet 6(8):597–610. https://doi.org/10.1038/nrg1655
Shah DK, Missmer SA, Berry KF, Racowsky C, Ginsburg ES (2011) Effect of obesity on oocyte and embryo quality in women undergoing in vitro fertilization. Obstet Gynecol 118(1):63–70. https://doi.org/10.1097/AOG.0b013e31821fd360
Snider AP, Wood JR (2019) Obesity induces ovarian inflammation and reduces oocyte quality. Reproduction 158(3):R79–R90. https://doi.org/10.1530/REP-18-0583
Sohrabi M, Roushandeh AM, Alizadeh Z, Vahidinia A, Vahabian M, Hosseini M (2015) Effect of a high fat diet on ovary morphology, in vitro development, in vitro fertilisation rate and oocyte quality in mice. Singap Med J 56(10):573–579. https://doi.org/10.11622/smedj.2015085
Sultan S, Patel AG, El-Hassani S, Whitelaw B, Leca BM, Vincent RP, le Roux CW, Rubino F, Aywlin SJB, Dimitriadis GK (2020) Male obesity associated gonadal dysfunction and the role of bariatric surgery. Front Endocrinol (Lausanne) 11:408. https://doi.org/10.3389/fendo.2020.00408
Turek-Plewa J, Jagodzinski PP (2005) The role of mammalian DNA methyltransferases in the regulation of gene expression. Cell Mol Biol Lett 10(4):631–647
Tzika E, Dreker T, Imhof A (2018) Epigenetics and metabolism in health and disease. Front Genet 9:361. https://doi.org/10.3389/fgene.2018.00361
Uysal F, Ozturk S (2017) DNA methyltransferases in mammalian oocytes. Results Probl Cell Differ 63:211–222. https://doi.org/10.1007/978-3-319-60855-6_10
Uysal F, Akkoyunlu G, Ozturk S (2015) Dynamic expression of DNA methyltransferases (DNMTs) in oocytes and early embryos. Biochimie 116:103–113. https://doi.org/10.1016/j.biochi.2015.06.019
Uysal F, Akkoyunlu G, Ozturk S (2016) DNA methyltransferases exhibit dynamic expression during spermatogenesis. Reprod Biomed Online 33(6):690–702. https://doi.org/10.1016/j.rbmo.2016.08.022
Uysal F, Ozturk S, Akkoyunlu G (2017) DNMT1, DNMT3A and DNMT3B proteins are differently expressed in mouse oocytes and early embryos. J Mol Histol 48(5–6):417–426. https://doi.org/10.1007/s10735-017-9739-y
Uysal F, Akkoyunlu G, Ozturk S (2019) Decreased expression of DNA methyltransferases in the testes of patients with non-obstructive azoospermia leads to changes in global DNA methylation levels. Reprod Fertil Dev. https://doi.org/10.1071/RD18246
Uysal F, Sukur G, Cinar O (2022) DNMT enzymes differentially alter global DNA methylation in a stage-dependent manner during spermatogenesis. Andrologia. https://doi.org/10.1111/and.14357
van Dijk SJ, Tellam RL, Morrison JL, Muhlhausler BS, Molloy PL (2015) Recent developments on the role of epigenetics in obesity and metabolic disease. Clin Epigenetics 7:66. https://doi.org/10.1186/s13148-015-0101-5
Wang N, Luo LL, Xu JJ, Xu MY, Zhang XM, Zhou XL, Liu WJ, Fu YC (2014) Obesity accelerates ovarian follicle development and follicle loss in rats. Metabolism 63(1):94–103. https://doi.org/10.1016/j.metabol.2013.09.001
Wang S, Pan MH, Hung WL, Tung YC, Ho CT (2019) From white to beige adipocytes: therapeutic potential of dietary molecules against obesity and their molecular mechanisms. Food Funct 10(3):1263–1279. https://doi.org/10.1039/c8fo02154f
Yoder JA, Soman NS, Verdine GL, Bestor TH (1997) DNA (cytosine-5)-methyltransferases in mouse cells and tissues. Studies with a mechanism-based probe. J Mol Biol 270(3):385–395. https://doi.org/10.1006/jmbi.1997.1125
You D, Nilsson E, Tenen DE, Lyubetskaya A, Lo JC, Jiang R, Deng J, Dawes BA, Vaag A, Ling C, Rosen ED, Kang S (2017) Dnmt3a is an epigenetic mediator of adipose insulin resistance. eLife. https://doi.org/10.7554/eLife.30766
Acknowledgements
We thank Prof. Erkan Yilmaz, who helped with animal diet regime, from Ankara University Biotechnology Institute.
Funding
This study was financially supported by TUBITAK 220S736.
Author information
Authors and Affiliations
Contributions
All authors qualify for authorship by contributing substantially to this article. G.S., F.U. and O.C. developed the original concept of this study. G.S. and F.U. collected data and performed statistical analyses, and G.S. and O.C. wrote the manuscript. All authors have contributed to critical discussion, reviewed the final version of the article and approved it for publication.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflicts of interest.
Consent to participate
Not applicable.
Consent for publication
Not applicable.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Sukur, G., Uysal, F. & Cinar, O. High-fat diet induced obesity alters Dnmt1 and Dnmt3a levels and global DNA methylation in mouse ovary and testis. Histochem Cell Biol 159, 339–352 (2023). https://doi.org/10.1007/s00418-022-02173-2
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00418-022-02173-2