Ali TAŞKIN, Nilhan COŞKUN, Ahmet KOCABAY

Farklı Partenogenetik Aktivasyon Sürelerinin Fare Embriyo Gelişimi ve Kalitesi Üzerine Etkileri

Çalışmamızın amacı, partenogenetik aktivasyonda farklı aktivasyon sürelerinin in vitro embriyo gelişimi ve kalitesi üzerindeki etkilerinin araştırılmasıdır. Superovule B6D2F1 ırkı dişi farelere uygulanan insan koryonik gonadotropin (hCG) enjeksiyonundan 14 saat sonra oositler elde edildi ve 18 saat sonra partenogenetik aktivasyona başlandı. Oositler, 10 mM stronsiyum klorür (SrCl2) + 5 μg/mL-1 sitokalazin B (CB) + 5 nM trikostatin A (TSA) Ca 2+ içermeyen Chatot Ziomek Brinster (CZB) medyumu içerisinde 3, 4, 5 ve 6 saat bekletildi. Aktivasyon sonrası, embriyo kültür medyumu + TSA’da inkübatörde 37°C ve %5 CO2 ortamında 2 saat bekletildi. Son olarak, tüm embriyolar 120 saat süre ile kültüre edildi. Bu çalışmadan elde edilen sonuçlar göre, 6 saatlik partenogenetik aktivasyon başarısının, 3, 4 ve 5 saatlik sürelere göre daha yüksek olduğu saptandı.

Anahtar Kelimeler:

Fare, Oosit, Partenogenetik, Aktivasyon Süresi

Effects of Different Parthenogenetic Activation Periods on Mouse Embryo Development and Quality

In the present study we investigate the effects of parthenogenetic activation on in vitro embryo development and quality in different activation periods. oocytes were obtained 14 hours after human chorionic gonadotropin (hCG) injection from superovulated B6D2F1 female mice then parthenogenetic activation started 18 hours after hCG injection. The oocytes were activated at different activation periods for 3, 4, 5 or 6 hours in 10 mM strontium chloride (SrCl2) + 5 μg/mL-1 Cytohalasine B (CB) + 5 nM Trichostatin A (TSA) containing a Ca 2+ free Chatot Ziomek Brinster (CZB) activation medium, followed by further incubation for two hours at 37°C and 5% CO2 in embryo culturing medium + TSA. The results in the present study suggested that the parthenogenetic activation of the 6 hour activation period was found to be higher than at 3, 4 and 5 hours.Keywords: Mouse, Oocyte, Parthenogenetic, Activation period

Keywords:

Mouse, Oocyte, Parthenogenetic, Activation period,

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Alberio R, Zakhartchenko V, Motlik J, Wolf E. Mammalian oocyte activation: lessons from the sperm and implications for nuclear transfer. Int J. Dev. Biol. 2001;45(7): 797-809.
Bai GY, Song SH, Wang ZD, Shan ZY, Sun RZ et al. Embryos aggregation improves development and imprinting gene expression in mouse parthenogenesis. Dev Growth Differ. 2016;58 (3): 270-279.
Campbell KHS. Nuclear equivalence, nuclear transfer, and the cell cycle. Cloning. 1999; 1(1): 3–15.
Cuthbertson KS, Whittingham DG, Cobbold PH. Free Ca2+ increases in exponential phases during mouse oocyte activation. Nature. 1981; 294(5843): 754-757.
Dandekar PV, Glass RH. Development of mouse embryos in vitro is affected by strain and culture medium. Gamete Res. 1987;17(4): 279–285.
Daughtry B, Mitalipov S. Concise review: parthenote stem cells for regenerative medicine: genetic, epigenetic, and developmental features. Stem. Cells. Transl. Med. 2014;3(3): 290-298.
De Sousa PA, Dobrinsky JR, Zhu J, Archibald AL, Ainslie A et al. Somatic cell nuclear transfer in the pig: control of pronuclear formation and integration with improved methods for activation and maintenance of pregnancy. Biol. Reprod. 2002;66(3): 642–650.
Demir K, Can A, Ertürk E, Özdemirci S, Karacam, H et al. Effect of different activation techniques on immature and in vitro matured cat oocytes. Kafkas Univ. Vet. Fak. Derg. 2014;20: 565-570.
Didié M, Christalla P, Rubart M, Muppala V, Döker S et al. Parthenogenetic stem cells for tissue-engineered heart repair. J. Clin. Invest. 2013;123(3): 1285-1298.
Fulka H, Hirose M, Inoue K, Ogonuki N, Wakisaka N et al. Production of mouse embryonic stem cell lines from maturing oocytes by direct conversion of meiosis into mitosis. Stem Cells. 2011;29(3): 517-527.
Gao W, Yu X, Hao J, Wang L, Qi M et al. Ascorbic acid improves parthenogenetic embryo development through TET proteins in mice. Biosci. Rep. 2019;39 (1): 1-8.
Han BS, Gao JL. Effects of chemical combinations on the parthenogenetic activation of mouse oocytes. Exp. Ther. Med. 2013;5(5): 1281-1288.
Heytens E, Soleimani R, Lierman S, De Meester S, Gerris J et al. Effect of ionomycin on oocyte activation and embryo development in mouse. Reprod. Biomed. Online. 2008;17 (6): 764-771.
Kishikawa H, Wakayama T, Yanagimachi R. Comparison of oocyte-activating agents for mouse cloning. Cloning. 1999;1(3): 153–159.
Kline D. Activation of the mouse egg. Theriogenology. 1996; 45(1): 81–90.
Ma SF, Liu XY, Miao DQ, Han ZB, Zhang X et al. Parthenogenetic activation of mouse oocytes by strontium chloride: A search for the best conditions. Theriogenology. 2005;64(5): 1142–1157.
Mallol A, Santaló J, Ibáñez E. Comparison of three differential mouse blastocyst staining methods. Syst. Biol. Reprod. Med. 2013;59 (2): 117-122.
Mallol A, Santaló J, Ibáñez E. Psammaplin a improves development and quality of somatic cell nuclear transfer mouse embryos. Cell Reprogram. 2014;16 (5): 392-406.
Sung LY, Chang CC, Amano T, Lin CJ, Amano M et al. Efficient derivation of embryonic stem cells from nuclear transfer and parthenogenetic embryos derived from cryopreserved oocytes. Cell Reprogram. 2010;12 (2): 203-211.
Taşkın AC, Kocabay A, Ebrahimi A, Karahüseyinoğlu S, Şahin GN et al. Leptin treatment of in vitro cultured embryos increases outgrowth rate of inner cell mass during embryonic stem cell derivation. In Vitro Cell Dev. Biol. Anim. 2019a;55 (7): 473-481.
Taşkın AC, Kocabay A. Leptin supplementation in embryo culture medium increases in vivo implantation rates in mice. Turk J Vet Anim Sci.2019b;43 (3): 359-363.
Versieren K, Heindryckx B, Lierman S, Gerris J, De Sutter P. Developmental competence of parthenogenetic mouse and human embryos after chemical or electrical activation. Reprod Biomed Online.2010;21(6): 769-775.
Wakayama T, Perry AC, Zuccotti M, Johnson KR, Yanagimachi R. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature. 1998;394 (6691): 369-374.