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Selection of developmentally competent immature equine oocytes with brilliant cresyl blue stain prior to in vitro maturation with equine growth hormone

Published online by Cambridge University Press:  01 February 2013

Gabriel R. Pereira ,
Pedro L. Lorenzo ,
Gustavo F. Carneiro ,
Sylvie Bilodeau-Goeseels ,
John P. Kastelic ,
Alejandro Esteller-Vico ,
Manel Lopez-Bejar  and
Irwin K.M. Liu
Gabriel R. Pereira*
Affiliation:
Federal University of Pelotas – UFPEL, School of Veterinary Medicine, Campus Capão do Leão, s/n° – Mailbox 354, Zip 96010–900, Pelotas, RS, Brasil. Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
Pedro L. Lorenzo
Affiliation:
Animal Physiology Department, Veterinary School, Universidad Complutense de Madrid, Madrid 28040, Spain.
Gustavo F. Carneiro
Affiliation:
Animal Reproduction Department, Federal Rural University of Pernambuco, Garanhuns, PE 52296–901, Brazil.
Sylvie Bilodeau-Goeseels
Affiliation:
Lethbridge Research Centre, P.O. Box 3000, 5403 1st Avenue S., Lethbridge, AB T1J4B1, Canada.
John P. Kastelic
Affiliation:
Lethbridge Research Centre, P.O. Box 3000, 5403 1st Avenue S., Lethbridge, AB T1J4B1, Canada.
Alejandro Esteller-Vico
Affiliation:
Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
Manel Lopez-Bejar
Affiliation:
Departament de Sanitat i d'Anatomia Animals, Autonomous University of Barcelona, Edifici V 08193 Bellaterra, Barcelona, Spain.
Irwin K.M. Liu
Affiliation:
Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
*
All correspondence to: Gabriel Ribas Pereira. Federal University of Pelotas – UFPEL, School of Veterinary Medicine, Campus Capão do Leão, s/n° – Mailbox 354, Zip 96010–900, Pelotas, RS, Brasil. Tel: +55 55 99972240 (Mobile) or +55 53 3275 7188 (Laboratory). E-mail: [email protected] or [email protected]
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Summary

Immature oocytes synthesize a variety of proteins that include the enzyme glucose-6-phosphate dehydrogenase (G6PDH). Brilliant cresyl blue (BCB) is a vital blue dye that assesses intracellular activity of G6PDH, an indirect measure of oocyte maturation. The objective was to evaluate the BCB test as a criterion to assess developmental competence of equine oocytes and to determine if equine growth hormone (eGH) enhanced in vitro maturation (IVM) of equine oocyte. Cumulus–oocytes complexes (COCs) were recovered by aspirating follicles <30 mm in diameter from abattoir-derived ovaries and were evaluated morphologically. Thereafter, COCs were exposed to BCB (26 μM) for 90 min at 39°C and selected based on the colour of their cytoplasm (BCB positive/BCB+ or BCB negative/BCB–). The COCs were allocated as follows: (a) IVM medium; (b) eGH group; (c) BCB–/IVM; (d) BCB+/IVM; (e) BCB–/eGH; and (f) BCB+/eGH. Then, COCs were cultured in vitro for 30 h, at 39°C in a 5%CO2 humidified air atmosphere. Cumulus-free oocytes were incubated in 10 μg/ml of bis-benzamide for 20 min at 39°C and nuclear maturation was evaluated with epifluorescence microscopy. Of the 39 COCs selected morphologically and subjected to BCB staining, 18/39 (46.2%) were classified as BCB+ and 21/39 (53.8%) as BCB– (P > 0.05). Maturation was not affected significantly by BCB classification, but the maturation rate was higher for oocytes that had been exposed to exogenous eGH versus controls (16/28, 57.1% versus 8/26, 30.8%, P < 0.05). In the present study, the BCB test was not useful for predicting competent equine oocytes prior to IVM. However, eGH enhanced equine oocyte maturation in vitro.

Keywords

Brilliant cresyl blueEquine growth hormoneHorseIn vitro maturationOocyte competence
Type
Research Article
Information
Zygote , Volume 22 , Issue 4 , November 2014 , pp. 500 - 504
Copyright
Copyright © Cambridge University Press 2013 

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References

Alm, H., Torner, H., Lohrke, B., Viergutz, T., Ghoneim, I.M. & Kanitz, W. (2005). Bovine blastocyst development rate in vitro is influenced by selection of oocytes by brilliant cresyl blue staining before IVM as indicator for glucose-6-phosphate dehydrogenase activity. Theriogenology 63, 2194–205.CrossRefGoogle ScholarPubMed
Del Campo, M.R., Donoso, M.X., Parrish, J.J. & Ginther, O.J. (1990). In vitro fertilization and in vitro matured equine oocytes. Equine Vet. Sci. 10, 18–22.CrossRefGoogle Scholar
Dell'Aquila, M.E., Fusco, S., Lacandra, G.M. & Maritato, F. (1997). Intracytoplasmic sperm injection (ICSI) versus conventional IVF on abattoir-derived and in vitro matured oocytes. Theriogenology 47, 1139–56.CrossRefGoogle Scholar
Ericsson, S.A., Boyce, M.L., Funahashi, H. & Day, B.N. (1993). Assessment of porcine oocytes using brilliant cresyl blue. Theriogenology 39, 214.CrossRefGoogle Scholar
Goovaerts, I.G.F., Leroy, J.L.M.R., Jorssen, E.P.A. & Bols, P.E.J. (2010). Noninvasive bovine oocyte quality assessment: possibilities of a single oocyte culture. Theriogenology 74, 1509–20.CrossRefGoogle ScholarPubMed
Goudet, G., Bézard, J., Duchamp, G., Gerard, N. & Palmer, E. (1997). Equine oocyte competence for nuclear and cytoplasmic in vitro maturation: effects of follicle size and hormonal environment. Biol. Reprod. 57, 232–45.CrossRefGoogle ScholarPubMed
Hinrichs, K. & Schmidt, A.L. (2000). Meiotic competence in horse oocytes: interactions among chromatin configuration, follicle size, cumulus morphology, and season. Biol. Reprod. 62, 1402–8.CrossRefGoogle ScholarPubMed
Ishizaki, C., Watanabe, H., Bhuiyan, M.M.U. & Fukui, Y. (2009). Developmental competence of porcine oocytes selected by brilliant cresyl blue and matured individually in a chemically defined culture medium. Theriogenology 72, 72–80.CrossRefGoogle Scholar
Izadyar, F., Colenbrander, B. & Bevers, M.M. (1996). In vitro maturation of bovine oocytes in the presence of growth hormone accelerates nuclear maturation and promotes subsequent embryonic development. Mol. Reprod. Dev. 45, 372–7.Google ScholarPubMed
Izadyar, F., Hage, W.J., Colenbrander, B. & Bevers, M.M. (1998). The promotory effect of growth hormone on the developmental competence of in vitro matured bovine oocytes is due to improved cytoplasmic maturation. Mol. Reprod. Dev. 49, 444–53.3.0.CO;2-U>CrossRefGoogle ScholarPubMed
Manjunatha, B.M., Gupta, P.S.P., Devaraj, M., Ravindra, J.P. & Nandi, S. (2007). Selection of developmentally competent buffalo oocytes by brilliant cresyl blue staining before IVM. Theriogenology 68, 1299–304.Google ScholarPubMed
Marchal, R., Caillaud, M., Martoriati, A., Gerard, N., Mermillod, P. & Goudet, G. (2003). Effect of growth hormone (GH) on in vitro nuclear and cytoplasmic oocyte maturation, cumulus expansion, hyaluronan synthases, and connexins 32 and 43 expression, and GH receptor messenger RNA expression in equine and porcine species. Biol. Reprod. 69, 1013–22.CrossRefGoogle ScholarPubMed
Opiela, J., Katska-Ksiazkiewicz, L., Lipinski, D., Słomski, R., Bzowska, M. & Rynska, B. (2008). Interactions among activity of glucose-6-phosphate dehydrogenase in immature oocytes, expression of apoptosis-related genes Bcl-2 and Bax, and developmental competence following IVP in cattle. Theriogenology 69, 546–55.CrossRefGoogle ScholarPubMed
Pereira, G.R., Lorenzo, P.L., Carneiro, G.F., Ball, B.A., Goncalves, P.B., Pegoraro, L.M., Bilodeau-Goeseels, S., Kastelic, J.P., Casey, P.J. & Liu, I.K.M. (2012). The effect of growth hormone (GH) and insulin-like growth factor-I (IGF-I) on in vitro maturation of equine oocytes. Zygote 20, 353–60.CrossRefGoogle ScholarPubMed
Pereira, G.R., Lorenzo, P.L., Carneiro, G.F., Bilodeau-Goeseels, S., Kastelic, J.P., Pegoraro, L.M., Pimentel, C.A., Esteller-Vico, A., Illera, J.C., Silvan, G., Casey, P.J. & Liu, I.K.M. (2006). Effect of equine growth hormone on in vitro maturation and steroid-levels response of equine oocytes. Anim. Reprod. Sci. 94, 364–5.Google Scholar
Pujol, M., Lopez-Bejar, M. & Paramio, M.T. (2004). Developmental competence of heifer oocytes selected using the brilliant cresyl blue (BCB) test. Theriogenology 61, 735–44.Google ScholarPubMed
Roca, J., Martinez, E., Vazquez, J.M. & Lucas, X. (1998). Selection of immature pig oocytes for homologous in vitro penetration assays with the brilliant cresyl blue test. Reprod. Fertil. Dev. 10, 479–85.CrossRefGoogle ScholarPubMed
Rodrigues, B.A., Rodriguez, P., Silva, A.E.F., Cavalcante, L.F., Fletrin, C. & Rodrigues, J.L. (2009). Preliminary study in immature canine oocytes stained with brilliant cresyl blue and obtained from bitches with low and high progesterone profiles. Reprod. Dom. Anim. 44, 255–8.CrossRefGoogle Scholar
Rodriguez-Gonzalez, E., Lopez-Bejar, M., Velilla, E. & Paramio, M.T. (2002). Selection of prepubertal goat oocytes using the brilliant cresyl blue test. Theriogenology 57, 1397–409.CrossRefGoogle ScholarPubMed