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Journal of Assisted Reproduction and Genetics

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01-04-2015 | Gamete Biology

Effect of L-carnitine supplementation on maturation and early embryo development of immature mouse oocytes selected by brilliant cresyle blue staining

Authors: Zohreh Zare, Reza Masteri Farahani, Mohammad Salehi, Abbas Piryaei, Marefat Ghaffari Novin, Fatemeh Fadaei Fathabadi, Moslem Mohammadi, Maryam Dehghani-Mohammadabadi

Published in: Journal of Assisted Reproduction and Genetics | Issue 4/2015

Abstract

Purpose

The present study was designed to investigate the effect of L-carnitine treatment during IVM on nuclear and cytoplasmic maturation of immature oocytes selected by Brilliant Cresyle Blue (BCB) staining, and their subsequent developmental competence.

Materials & methods

Compact cumulus-oocyte complexes (COCs) were collected from NMRI mice ovaries and stained with BCB staining. BCB+ (colored cytoplasm) oocytes were then cultured in tissue culture medium (TCM) 199 with 0.0, 0.3 and 0.6 mg/ml L-carnitine.

Results

The both L-carnitine concentrations significantly increased the intracellular glutathione (P < 0.001), nuclear maturation (P < 0.01) and expression levels of cyclin-dependent kinase1 (CDK1) (P < 0.05). Moreover, treated oocytes with 0.6 mg/ml L-carnitine showed increased (P < 0.05) expression of mitogen-activated protein kinase1 (MAPK1) mRNA. Also, adding L-carnitine (0.6 mg/ml) to IVM medium significantly increased the cleavage rate (P < 0.05). The blastocyst development rate (BDR) in the both L-carnitine treated groups was significantly higher (P < 0.001) than the control group. L-carnitine had no significant effect on total blastocyst cell numbers.

Conclusions

These data indicated that L-carnitine supplementation during IVM of immature BCB+ oocytes improved preimplantation developmental competence of oocytes after IVF, probably by accelerating cytoplasmic and nuclear maturation of oocytes. It may provide a novel approach to improving ART outcomes in infertile couples.

Albuz FK, Sasseville M, Lane M, Armstrong DT, Thompson JG, Gilchrist RB. Simulated physiological oocyte maturation (SPOM): a novel in vitro maturation system that substantially improves embryo yield and pregnancy outcomes. Hum Reprod. 2010;25:2999–3011.CrossRefPubMed

Whelan 3rd JG, Vlahos NF. The ovarian hyperstimulation syndrome. Fertil Steril. 2000;73:883–96.CrossRefPubMed

Wang N, Le F, Zhan QT, Li L, Dong MY, Ding GL, et al. Effects of in vitro maturation on histone acetylation in metaphase II oocytes and early cleavage embryos. Obstet Gynecol Int. 2010;2010:989278.PubMedCentralPubMed

Baart EB, Martini E, Eijkemans MJ, Van Opstal D, Beckers NG, Verhoeff A, et al. Milder ovarian stimulation for in-vitro fertilization reduces aneuploidy in the human preimplantation embryo: a randomized controlled trial. Hum Reprod. 2007;22:980–8.CrossRefPubMed

Buckett WM, Chian RC, Holzer H, Dean N, Usher R, Tan SL. Obstetric outcomes and congenital abnormalities after in vitro maturation, in vitro fertilization, and intracytoplasmic sperm injection. Obstet Gynecol. 2007;110:885–91.CrossRefPubMed

Nagai T. The improvement of in vitro maturation systems for bovine and porcine oocytes. Theriogenology. 2001;55:1291–301.CrossRefPubMed

Su J, Wang Y, Li R, Peng H, Hua S, Li Q, et al. Oocytes selected using BCB staining enhance nuclear reprogramming and the in vivo development of SCNT embryos in cattle. PLoS ONE. 2012;7:e36181.CrossRefPubMedCentralPubMed

Torner H, Ghanem N, Ambros C, Holker M, Tomek W, Phatsara C, et al. Molecular and subcellular characterisation of oocytes screened for their developmental competence based on glucose-6-phosphate dehydrogenase activity. Reproduction. 2008;135:197–212.CrossRefPubMed

Roca J, Martinez E, Vazquez JM, Lucas X. Selection of immature pig oocytes for homologous in vitro penetration assays with the brilliant cresyl blue test. Reprod Fertil Dev. 1998;10:479–85.CrossRefPubMed

10.

Catala MG, Izquierdo D, Uzbekova S, Morato R, Roura M, Romaguera R, et al. Brilliant Cresyl Blue stain selects largest oocytes with highest mitochondrial activity, maturation-promoting factor activity and embryo developmental competence in prepubertal sheep. Reproduction. 2011;142:517–27.CrossRefPubMed

11.

Alm H, Torner H, Lohrke B, Viergutz T, Ghoneim IM, Kanitz W. Bovine blastocyst development rate in vitro is influenced by selection of oocytes by brillant cresyl blue staining before IVM as indicator for glucose-6-phosphate dehydrogenase activity. Theriogenology. 2005;63:2194–205.CrossRefPubMed

12.

Karja NW, Wongsrikeao P, Murakami M, Agung B, Fahrudin M, Nagai T, et al. Effects of oxygen tension on the development and quality of porcine in vitro fertilized embryos. Theriogenology. 2004;62:1585–95.CrossRefPubMed

13.

Mo X, Wu G, Yuan D, Jia B, Liu C, Zhu S, et al. Leukemia inhibitory factor enhances bovine oocyte maturation and early embryo development. Mol Reprod Dev. 2014;81:608–18.CrossRefPubMed

14.

Choe C, Shin YW, Kim EJ, Cho SR, Kim HJ, Choi SH, et al. Synergistic effects of glutathione and beta-mercaptoethanol treatment during in vitro maturation of porcine oocytes on early embryonic development in a culture system supplemented with L-cysteine. J Reprod Dev. 2010;56:575–82.CrossRefPubMed

15.

Dehghani-Mohammadabadi M, Salehi M, Farifteh F, Nematollahi S, Arefian E, Hajjarizadeh A, et al. Melatonin modulates the expression of BCL-xl and improve the development of vitrified embryos obtained by IVF in mice. J Assist Reprod Genet. 2014;31:453–61.CrossRefPubMed

16.

Vanella A, Russo A, Acquaviva R, Campisi A, Di Giacomo C, Sorrenti V, et al. L -propionyl-carnitine as superoxide scavenger, antioxidant, and DNA cleavage protector. Cell Biol Toxicol. 2000;16:99–104.CrossRefPubMed

17.

Stojkovic M, Machado SA, Stojkovic P, Zakhartchenko V, Hutzler P, Goncalves PB, et al. Mitochondrial distribution and adenosine triphosphate content of bovine oocytes before and after in vitro maturation: correlation with morphological criteria and developmental capacity after in vitro fertilization and culture. Biol Reprod. 2001;64:904–9.CrossRefPubMed

18.

Gulcin I. Antioxidant and antiradical activities of L-carnitine. Life Sci. 2006;78:803–11.CrossRefPubMed

19.

Qi SN, Zhang ZF, Wang ZY, Yoshida A, Ueda T. L-carnitine inhibits apoptotic DNA fragmentation induced by a new spin-labeled derivative of podophyllotoxin via caspase-3 in Raji cells. Oncol Rep. 2006;15:119–22.PubMed

20.

Winter BK, Fiskum G, Gallo LL. Effects of L-carnitine on serum triglyceride and cytokine levels in rat models of cachexia and septic shock. Br J Cancer. 1995;72:1173–9.CrossRefPubMedCentralPubMed

21.

Abdelrazik H, Sharma R, Mahfouz R, Agarwal A. L-carnitine decreases DNA damage and improves the in vitro blastocyst development rate in mouse embryos. Fertil Steril. 2009;91:589–96.CrossRefPubMed

22.

Cook JA, Pass HI, Iype SN, Friedman N, DeGraff W, Russo A, et al. Cellular glutathione and thiol measurements from surgically resected human lung tumor and normal lung tissue. Cancer Res. 1991;51:4287–94.PubMed

23.

Agarwal A, Allamaneni SS, Said TM. Chemiluminescence technique for measuring reactive oxygen species. Reprod BioMed Online. 2004;9:466–8.CrossRefPubMed

24.

Sharma RK, Agarwal A. Role of reactive oxygen species in gynecologic diseases. Reprod Med Biol. 2004;3:177–99.CrossRef

25.

Zuccotti M, Boiani M, Ponce R, Guizzardi S, Scandroglio R, Garagna S, et al. Mouse Xist expression begins at zygotic genome activation and is timed by a zygotic clock. Mol Reprod Dev. 2002;61:14–20.CrossRefPubMed

26.

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;25:402–8.CrossRefPubMed

27.

Sirard MA, Richard F, Blondin P, Robert C. Contribution of the oocyte to embryo quality. Theriogenology. 2006;65:126–36.CrossRefPubMed

28.

Opiela J, Lipinski D, Slomski R, Katska-Ksiazkiewicz L. Transcript expression of mitochondria related genes is correlated with bovine oocyte selection by BCB test. Anim Reprod Sci. 2010;118:188–93.CrossRefPubMed

29.

Thiyagarajan B, Valivittan K. Ameliorating effect of vitamin E on in vitro development of preimplantation buffalo embryos. J Assist Reprod Genet. 2009;26:217–25.CrossRefPubMedCentralPubMed

30.

Kowaltowski AJ, Vercesi AE. Mitochondrial damage induced by conditions of oxidative stress. Free Radic Biol Med. 1999;26:463–71.CrossRefPubMed

31.

Agarwal A, Gupta S, Sharma RK. Role of oxidative stress in female reproduction. Reprod Biol Endocrinol. 2005;3:28.CrossRefPubMedCentralPubMed

32.

Shanti A, Santanam N, Morales AJ, Parthasarathy S, Murphy AA. Autoantibodies to markers of oxidative stress are elevated in women with endometriosis. Fertil Steril. 1999;71:1115–8.CrossRefPubMed

33.

Karuputhula NB, Chattopadhyay R, Chakravarty B, Chaudhury K. Oxidative status in granulosa cells of infertile women undergoing IVF. Syst Biol Reprod Med. 2013;59:91–8.CrossRefPubMed

34.

Natarajan R, Shankar MB, Munuswamy D. Effect of alpha-tocopherol supplementation on in vitro maturation of sheep oocytes and in vitro development of preimplantation sheep embryos to the blastocyst stage. J Assist Reprod Genet. 2010;27:483–90.CrossRefPubMedCentralPubMed

35.

Fenkci SM, Fenkci V, Oztekin O, Rota S, Karagenc N. Serum total L-carnitine levels in non-obese women with polycystic ovary syndrome. Hum Reprod. 2008;23:1602–6.CrossRefPubMed

36.

Wu GQ, Jia BY, Li JJ, Fu XW, Zhou GB, Hou YP, et al. L-carnitine enhances oocyte maturation and development of parthenogenetic embryos in pigs. Theriogenology. 2011;76:785–93.CrossRefPubMed

37.

You J, Lee J, Hyun SH, Lee E. L-carnitine treatment during oocyte maturation improves in vitro development of cloned pig embryos by influencing intracellular glutathione synthesis and embryonic gene expression. Theriogenology. 2012;78:235–43.CrossRefPubMed

38.

Rausell F, Pertusa JF, Gomez-Piquer V, Hermenegildo C, Garcia-Perez MA, Cano A, et al. Beneficial effects of dithiothreitol on relative levels of glutathione S-transferase activity and thiols in oocytes, and cell number, DNA fragmentation and allocation at the blastocyst stage in the mouse. Mol Reprod Dev. 2007;74:860–9.CrossRefPubMed

39.

Luberda Z. The role of glutathione in mammalian gametes. Reprod Biol. 2005;5:5–17.PubMed

40.

Yoshida M, Ishigaki K, Nagai T, Chikyu M, Pursel VG. Glutathione concentration during maturation and after fertilization in pig oocytes: relevance to the ability of oocytes to form male pronucleus. Biol Reprod. 1993;49:89–94.CrossRefPubMed

41.

Somfai T, Kaneda M, Akagi S, Watanabe S, Haraguchi S, Mizutani E, et al. Enhancement of lipid metabolism with L-carnitine during in vitro maturation improves nuclear maturation and cleavage ability of follicular porcine oocytes. Reprod Fertil Dev. 2011;23:912–20.CrossRefPubMed

42.

Jeulin C, Lewin LM. Role of free L-carnitine and acetyl-L-carnitine in post-gonadal maturation of mammalian spermatozoa. Hum Reprod Update. 1996;2:87–102.CrossRefPubMed

43.

Dunning KR, Cashman K, Russell DL, Thompson JG, Norman RJ, Robker RL. Beta-oxidation is essential for mouse oocyte developmental competence and early embryo development. Biol Reprod. 2010;83:909–18.CrossRefPubMed

44.

Zhou W, Xiang T, Walker S, Farrar V, Hwang E, Findeisen B, et al. Global gene expression analysis of bovine blastocysts produced by multiple methods. Mol Reprod Dev. 2008;75:744–58.CrossRefPubMed

45.

Racedo SE, Wrenzycki C, Lepikhov K, Salamone D, Walter J, Niemann H. Epigenetic modifications and related mRNA expression during bovine oocyte in vitro maturation. Reprod Fertil Dev. 2009;21:738–48.CrossRefPubMed

46.

Salhab M, Dhorne-Pollet S, Auclair S, Guyader-Joly C, Brisard D, Dalbies-Tran R, et al. In vitro maturation of oocytes alters gene expression and signaling pathways in bovine cumulus cells. Mol Reprod Dev. 2013;80:166–82.CrossRefPubMed

47.

Paradis F, Novak S, Murdoch GK, Dyck MK, Dixon WT, Foxcroft GR. Temporal regulation of BMP2, BMP6, BMP15, GDF9, BMPR1A, BMPR1B, BMPR2 and TGFBR1 mRNA expression in the oocyte, granulosa and theca cells of developing preovulatory follicles in the pig. Reproduction. 2009;138:115–29.CrossRefPubMed

48.

Lee SE, Kim EY, Choi HY, Moon JJ, Park MJ, Lee JB, et al. Rapamycin rescues the poor developmental capacity of aged porcine oocytes. Asian-Australas J Anim Sci. 2014;27:635–47.CrossRefPubMedCentralPubMed

49.

Hussein TS, Thompson JG, Gilchrist RB. Oocyte-secreted factors enhance oocyte developmental competence. Dev Biol. 2006;296:514–21.CrossRefPubMed

50.

Zhang DX, Park WJ, Sun SC, Xu YN, Li YH, Cui XS, et al. Regulation of maternal gene expression by MEK/MAPK and MPF signaling in porcine oocytes during in vitro meiotic maturation. J Reprod Dev. 2011;57:49–56.CrossRefPubMed

51.

De Smedt V, Poulhe R, Cayla X, Dessauge F, Karaiskou A, Jessus C, et al. Thr-161 phosphorylation of monomeric Cdc2. Regulation by protein phosphatase 2C in Xenopus oocytes. J Biol Chem. 2002;277:28592–600.CrossRefPubMed

52.

Kim KH, Kim EY, Kim Y, Kim E, Lee HS, Yoon SY, et al. Gas6 downregulation impaired cytoplasmic maturation and pronuclear formation independent to the MPF activity. PLoS ONE. 2011;6:e23304.CrossRefPubMedCentralPubMed

53.

Takahashi T, Inaba Y, Somfai T, Kaneda M, Geshi M, Nagai T, et al. Supplementation of culture medium with L-carnitine improves development and cryotolerance of bovine embryos produced in vitro. Reprod Fertil Dev. 2013;25:589–99.CrossRefPubMed

Title: Effect of L-carnitine supplementation on maturation and early embryo development of immature mouse oocytes selected by brilliant cresyle blue staining
Authors: Zohreh Zare
Reza Masteri Farahani
Mohammad Salehi
Abbas Piryaei
Marefat Ghaffari Novin
Fatemeh Fadaei Fathabadi
Moslem Mohammadi
Maryam Dehghani-Mohammadabadi
Publication date: 01-04-2015
Publisher: Springer US
Published in: Journal of Assisted Reproduction and Genetics / Issue 4/2015
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-015-0430-5

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Effect of L-carnitine supplementation on maturation and early embryo development of immature mouse oocytes selected by brilliant cresyle blue staining

Abstract

Purpose

Materials & methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Materials & methods

Results

Conclusions

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