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 STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Journal of Ovarian Research
Published in: Journal of Ovarian Research 1/2018

Open Access 01-12-2018 | Research

Retinoic acid enhances germ cell differentiation of mouse skin-derived stem cells

Authors: Paul W. Dyce, Neil Tenn, Gerald M. Kidder

Published in: Journal of Ovarian Research | Issue 1/2018

Login to get access

Abstract

Background

Retinoic acid (RA) signaling has been identified as a key driver in male and female gamete development. The presence of RA is a critical step in the initiation of meiosis and is required for the production of competent oocytes from primordial germ cells. Meiosis has been identified as a difficult biological process to recapitulate in vitro, when differentiating stem cells to germ cells. We have previously shown that primordial germ cell-like cells, and more advanced oocyte-like cells (OLCs), can be formed by differentiating mouse skin-derived stem cells. However, the OLCs remain unable to function due to what appears to be failure of meiotic initiation. The aim of this study was to determine the effect of RA treatment, during stem cell differentiation to germ cells, particularly on the initiation of meiosis.

Results

Using qPCR we found significant increases in the meiosis markers Stra8 and Sycp3 and a significant reduction in the meiosis inhibitor Nanos2, in the differentiating populations. Furthermore, OLCs from the RA treated group, expressed significantly more of the meiosis regulatory gene Marf1 and the oocyte marker Oct4. At the protein level RA treatment was found to increase the expression of the gap junction protein CX43 and the pluripotency marker OCT4. Moreover, the expression of SYCP3 was significantly upregulated and the localization pattern better matched that of control fetal ovarian cells. RA treatment also improved the structural integrity of the OLCs produced by initiating the expression of all three zona pellucida transcripts (Zp1–3) and improving ZP3 expression levels and localization. Finally, the addition of RA during differentiation led to an almost two-fold increase in the number of OLCs recovered and increased their in vitro growth.

Conclusion

RA is a key driver in the formation of functioning gametes and its addition during stem cell to germ cell differentiation improves OLCs entry into meiosis.
Literature
1.
Speed RM. Meiosis in the foetal mouse ovary. I. An analysis at the light microscope level using surface-spreading. Chromosoma. 1982;85:427–37.CrossRefPubMed
2.
Bowles J, Knight D, Smith C, Wilhelm D. Retinoid signaling determines germ cell fate in mice. Science. 2006;312(5773):596–600.CrossRefPubMed
3.
Baltus AE, Menke DB, Hu Y-CC, Goodheart ML, Carpenter AE, de Rooij DG, Page DC. In germ cells of mouse embryonic ovaries, the decision to enter meiosis precedes premeiotic DNA replication. Nat Genet. 2006;38:1430–4.CrossRefPubMed
4.
Menke DB, Koubova J, Page DC. Sexual differentiation of germ cells in XX mouse gonads occurs in an anterior-to-posterior wave. Dev Biol. 2003;262:303–12.CrossRefPubMed
5.
White JA, Ramshaw H, Taimi M, Stangle W, Zhang A, Everingham S, Creighton S, Tam SP, Jones G, Petkovich M. Identification of the human cytochrome P450, P450RA1-2, which is predominantly expressed in the adult cerebellum and is responsible for all-trans-retinoic acid metabolism. Proc Natl Acad Sci. 2000;97:6403–8.CrossRefPubMedPubMedCentral
6.
Zhou Q, Li Y, Nie R, Friel P, Mitchell D, Evanoff RM, Pouchnik D, Banasik B, McCarrey JR, Small C, Griswold MD. Expression of stimulated by retinoic acid gene 8 (Stra8) and maturation of murine gonocytes and spermatogonia induced by retinoic acid in vitro. Biol Reprod. 2008;78:537–45.CrossRefPubMed
7.
Adams IR, McLaren A. Sexually dimorphic development of mouse primordial germ cells: switching from oogenesis to spermatogenesis. Development. 2002;129(5):1155–64.PubMed
8.
9.
Li H, Clagett-Dame M. Vitamin a deficiency blocks the initiation of meiosis of germ cells in the developing rat ovary in vivo. Biol Reprod. 2009;81(5):996–1001.CrossRefPubMed
10.
Raverdeau M, Gely-Pernot A, Féret B. Retinoic acid induces Sertoli cell paracrine signals for spermatogonia differentiation but cell autonomously drives spermatocyte meiosis. Proc Natl Acad Sci. 2012;109(41):16582–7.CrossRefPubMedPubMedCentral
11.
Farini D, Scaldaferri ML, Iona S, Sala LG. Growth factors sustain primordial germ cell survival, proliferation and entering into meiosis in the absence of somatic cells. Dev Biol. 2005;285(1):49–56.CrossRefPubMed
12.
Koshimizu U, Watanabe M, Nakatsuji N. Retinoic acid is a potent growth activator of mouse primordial germ cells in vitro. Dev Biol. 1995;168(2):683–5.CrossRefPubMed
13.
Hübner K, Fuhrmann G, Christenson LK, Kehler J, Reinbold R, De La Fuente R, Wood J, Strauss JF, Boiani M, Schöler HR. Derivation of oocytes from mouse embryonic stem cells. Science. 2003;300:1251–6.CrossRefPubMed
14.
Geijsen N, Horoschak M, Kim K, Gribnau J, Eggan K, Daley GQ. Derivation of embryonic germ cells and male gametes from embryonic stem cells. Nature. 2004;427:148–54.CrossRefPubMed
15.
16.
17.
Dyce PW, Wen L, Li J. In vitro germline potential of stem cells derived from fetal porcine skin. Nat Cell Biol. 2006;8:384–90.CrossRefPubMed
18.
Dyce PW. Differentiation of newborn mouse skin derived stem cells into germ-like cells in vitro. J Vis Exp. 2013;77:e50486.
19.
Dyce PW, Liu J, Tayade C, Kidder GM, Betts DH, Li J. In vitro and in vivo germ line potential of stem cells derived from newborn mouse skin. PLoS One. 2011;6(5):e20339.CrossRefPubMedPubMedCentral
20.
Dyce PW, Shen W, Huynh E, Shao H, Villagómez DA, Kidder GM, King WA, Li J. Analysis of oocyte-like cells differentiated from porcine fetal skin-derived stem cells. Stem Cells Dev. 2011;20:809–19.CrossRefPubMed
21.
Ge W, Ma H-GG, Cheng S-FF, Sun Y-CC, Sun L-LL, Sun X-FF, Li L, Dyce P, Li J, Shi Q-HH, Shen W. Differentiation of early germ cells from human skin-derived stem cells without exogenous gene integration. Sci Rep. 2015;5:13822.CrossRefPubMedPubMedCentral
22.
Nayernia K, Nolte J, Michelmann HW, Lee JH. In vitro-differentiated embryonic stem cells give rise to male gametes that can generate offspring mice. Dev Cell. 2006;11(1):125–32.CrossRefPubMed
23.
Zhou Q, Wang M, Yuan Y, Wang X, Fu R, Wan H, Xie M, Liu M, Guo X, Zheng Y, Feng G, Shi Q, Zhao X-YY, Sha J, Zhou Q. Complete meiosis from embryonic stem cell-derived germ cells in vitro. Cell Stem Cell. 2016;18:330–40.CrossRefPubMed
24.
Toma J, Akhavan M, Fernandes K, Barnabé-Heider F, Sadikot A, Kaplan D, Miller F. Isolation of multipotent adult stem cells from the dermis of mammalian skin. Nat Cell Biol. 2001;3:778–84.CrossRefPubMed
25.
Winterhager E, Kidder GM. Gap junction connexins in female reproductive organs: implications for women's reproductive health. Hum Reprod Update. 2015;21(3):340–52.CrossRefPubMed
26.
Ge W, Cheng SF, Dyce PW, De Felici M, Shen W. Skin-derived stem cells as a source of primordial germ cell-and oocyte-like cells. Cell Death Dis. 2016;7(11):e2471.CrossRefPubMedPubMedCentral
27.
Semba RD (2012). On the “discovery” of vitamin a. Annals of nutrition and metabolism.
28.
29.
30.
Su Q, Sugiura S, Pendola C, Handel S, Eppig. MARF1 regulates essential Oogenic processes in mice. Science. 2012;335:14961499.CrossRef
31.
Su Y-Q, Sun F, Handel M, Schimenti J, Eppig J. Meiosis arrest female 1 (MARF1) has nuage-like function in mammalian oocytes. Proc Natl Acad Sci. 2012;109:18653–60.CrossRefPubMedPubMedCentral
32.
Liang L, Soyal SM, Dean J. FIGalpha, a germ cell specific transcription factor involved in the coordinate expression of the zona pellucida genes. Development. 1997;124(24):4939–47.PubMed
Metadata
Title
Retinoic acid enhances germ cell differentiation of mouse skin-derived stem cells
Authors
Paul W. Dyce
Neil Tenn
Gerald M. Kidder
Publication date
01-12-2018
Publisher
BioMed Central
Published in
Journal of Ovarian Research / Issue 1/2018
Electronic ISSN: 1757-2215
DOI
https://doi.org/10.1186/s13048-018-0390-3

Other articles of this Issue 1/2018

Journal of Ovarian Research 1/2018 Go to the issue

Research

Is the ovarian reserve influenced by vitamin D deficiency and the dress code in an infertile Iranian population?

Research

Mitochondrial membrane depolarization enhances TRAIL-induced cell death in adult human granulosa tumor cells, KGN, through inhibition of BIRC5

Research

MR findings of primary ovarian granulosa cell tumor with focus on the differentiation with other ovarian sex cord-stromal tumors

Research

Ovarian teratoma in children: a plea for collaborative clinical study

Research

Clinical study of a CT evaluation model combined with serum CA125 in predicting the treatment of newly diagnosed advanced epithelial ovarian cancer

Research

Gonadotropin and steroid hormones regulate pluripotent very small embryonic-like stem cells in adult mouse uterine endometrium