Skip to main content Accessibility help

Linolenic acid improves oocyte developmental competence and decreases apoptosis of in vitro-produced blastocysts in goat

  • Arash Veshkini (a1) (a2), Ali Akbar Khadem (a1), Abdollah Mohammadi-Sangcheshmeh (a3) (a1), Ali Asadi Alamouti (a1), Masoud Soleimani (a4) and Eduardo L. Gastal (a5)...


The effects of α-linolenic acid (ALA) on developmental competence of oocytes in goats were evaluated in this study. Initially, the level of ALA in small and large antral follicles was determined to be in a range of 0.018–0.028 mg/ml (64.6–100.6 μM, respectively). In vitro maturation was performed in the presence of various concentrations (10, 50, 100, or 200 μM) of ALA. Cumulus expansion, meiotic maturation, levels of intracellular glutathione (GSH), embryonic cleavage, blastocyst formation following parthenogenetic activation (PA) and in vitro fertilization (IVF), number of total and apoptotic cells in blastocyst, and expression of Bax, Bcl-2, and p53 genes in blastocyst cells were determined. Compared with the control, no improvement was observed in cumulus expansion in ALA-treated groups. At 50 μM concentration, ALA increased meiotic maturation rate but had no effect on GSH level. When oocytes treated with 50 μM ALA were subsequently used for PA or IVF, a higher rate of blastocyst formation was observed, and these embryos had a higher total cell number and a lower apoptotic cell number. Expression analyses of genes in blastocysts revealed lesser transcript abundances for Bax gene, and higher transcript abundances for Bcl-2 gene in 50 μM ALA group. Expression of p53 gene was also less observed in ALA-treated blastocysts. Our results show that ALA treatment at 50 μM during in vitro maturation (IVM) had a beneficial effect on maturation of goat oocytes and this, in turn, stimulated embryonic development and regulated apoptotic gene expression.


Corresponding author

All correspondence to: Abdollah Mohammadi-Sangcheshmeh. Department of Animal and Poultry Science, College of Aburaihan, University of Tehran, Pakdasht, Theran P.O.Box: 11365/7117, Iran. Fax: +98 21 36040907. E-mail:


Hide All
Abayasekara, D.R. & Wathes, D.C. (1999). Effects of altering dietary fatty acid composition on prostaglandin synthesis and fertility. Prostag. Leukot. Ess. 61, 275–87.
Abazari-Kia, A.H., Mohammadi-Sangcheshmeh, A., Dehghani-Mohammadabadi, M., Jamshidi-Adegani, F., Veshkini, A., Zhandi, M., Cinar, M.U. & Salehi, M. (2014). Intracellular glutathione content, developmental competence and expression of apoptosis-related genes associated with G6PDH-activity in goat oocyte. J. Assist. Reprod Genet. 31, 313–21.
Al Darwich, A., Perreau, C., Petit, M.H., Papillier, P., Dupont, J., Guillaume, D., Mermillod, P. & Guignot, F. (2010). Effect of PUFA on embryo cryoresistance, gene expression and AMPKalpha phosphorylation in IVF-derived bovine embryos. Prostag. Oth. Lipid M. 93, 30–6.
Amundson, S.A., Myers, T.G. & Fornace, A.J. Jr. (1998). Roles for p53 in growth arrest and apoptosis: putting on the brakes after genotoxic stress. Oncogene 17, 3287–99.
Andrade, L.N., de Lima, T.M., Curi, R. & Castrucci, A.M. (2005). Toxicity of fatty acids on murine and human melanoma cell lines. Toxicol. In Vitro 19, 553–60.
Barcelo-Coblijn, G. & Murphy, E.J. (2009). Alpha-linolenic acid and its conversion to longer chain n-3 fatty acids: benefits for human health and a role in maintaining tissue n-3 fatty acid levels. Prog. Lipid Res. 48, 355–74.
Bender, K., Walsh, S., Evans, A.C., Fair, T. & Brennan, L. (2010). Metabolite concentrations in follicular fluid may explain differences in fertility between heifers and lactating cows. Reproduction 139, 1047–55.
Browne, R.W. & Armstrong, D. (2000). HPLC analysis of lipid-derived polyunsaturated fatty acid peroxidation products in oxidatively modified human plasma. Clin. Chem. 46, 829–36.
Burlacu, A. (2003). Regulation of apoptosis by Bcl-2 family proteins. J. Cell. Mol. Med. 7, 249–57.
Cha, K.Y. & Chian, R.C. (1998). Maturation in vitro of immature human oocytes for clinical use. Hum. Reprod. Update 4, 103–20.
Childs, S., Hennessy, A.A., Sreenan, J.M., Wathes, D.C., Cheng, Z., Stanton, C., Diskin, M.G. & Kenny, D.A. (2008). Effect of level of dietary n-3 polyunsaturated fatty acid supplementation on systemic and tissue fatty acid concentrations and on selected reproductive variables in cattle. Theriogenology 70, 595611.
Chrenek, P., Kubovicova, E., Olexikova, L., Makarevich, A.V., Toporcerova, S. & Ostro, A. (2014). Effect of body condition and season on yield and quality of in vitro produced bovine embryos. Zygote Epub ahead of print.
Coyral-Castel, S., Rame, C., Fatet, A. & Dupont, J. (2010). Effects of unsaturated fatty acids on progesterone secretion and selected protein kinases in goat granulosa cells. Domest. Anim. Endocrinol. 38, 272–83.
Elvin, J.A., Yan, C. & Matzuk, M.M. (2000). Growth differentiation factor-9 stimulates progesterone synthesis in granulosa cells via a prostaglandin E2/EP2 receptor pathway. Proc. Natl. Acad. Sci. USA 97, 10288–93.
Fouladi-Nashta, A.A., Wonnacott, K.E., Gutierrez, C.G., Gong, J.G., Sinclair, K.D., Garnsworthy, P.C. & Webb, R. (2009). Oocyte quality in lactating dairy cows fed on high levels of n-3 and n-6 fatty acids. Reproduction 138, 771–81.
Funston, R.N. (2004). Fat supplementation and reproduction in beef females. J. Anim. Sci. 82 (E-Suppl.), E154–61.
Ghaffarilaleh, V., Fouladi-Nashta, A. & Paramio, M.T. (2014). Effect of alpha-linolenic acid on oocyte maturation and embryo development of prepubertal sheep oocytes. Theriogenology 82, 686–96.
Gladine, C., Rock, E., Morand, C., Bauchart, D. & Durand, D. (2007). Bioavailability and antioxidant capacity of plant extracts rich in polyphenols, given as a single acute dose, in sheep made highly susceptible to lipoperoxidation. Br. J. Nutr. 98, 691701.
Gross, A., McDonnell, J.M. & Korsmeyer, S.J. (1999). BCL-2 family members and the mitochondria in apoptosis. Genes Dev. 13, 1899–911.
Hao, Y., Lai, L., Mao, J., Im, G.S., Bonk, A. & Prather, R.S. (2003). Apoptosis and in vitro development of preimplantation porcine embryos derived in vitro or by nuclear transfer. Biol. Reprod. 69, 501–7.
Harrington, N.P., Surujballi, O.P., Waters, W.R. & Prescott, J.F. (2007). Development and evaluation of a real-time reverse transcription-PCR assay for quantification of gamma interferon mRNA to diagnose tuberculosis in multiple animal species. Clin. Vaccine Immunol. 14, 1563–71.
Korkmaz, C., Tekin, Y.B., Sakinci, M. & Ercan, C.M. (2015). Effects of maternal ageing on ICSI outcomes and embryo development in relation to oocytes morphological characteristics of birefringent structures. Zygote 23, 550–5.
Lan, G.C., Han, D., Wu, Y.G., Han, Z.B., Ma, S.F., Liu, X.Y., Chang, C.L. & Tan, J.H. (2005). Effects of duration, concentration, and timing of ionomycin and 6-dimethylaminopurine (6-DMAP) treatment on activation of goat oocytes. Mol. Reprod. Dev. 71, 380–8.
Lasiene, K., Vitkus, A., Valanciute, A. & Lasys, V. (2009). Morphological criteria of oocyte quality. Medicina (Kaunas) 45, 509–15.
Leroy, J.L., Vanholder, T., Mateusen, B., Christophe, A., Opsomer, G., de Kruif, A., Genicot, G. & Van Soom, A. (2005). Non-esterified fatty acids in follicular fluid of dairy cows and their effect on developmental capacity of bovine oocytes in vitro . Reproduction 130, 485–95.
Lussier, J.G., Matton, P., Guilbault, L.A., Grasso, F., Mapletoft, R.J. & Carruthers, T.D. (1994). Ovarian follicular development and endocrine responses in follicular-fluid-treated and hemi-ovariectomized heifers. J. Reprod. Fertil. 102, 95105.
Marei, W.F., Wathes, D.C. & Fouladi-Nashta, A.A. (2009). The effect of linolenic acid on bovine oocyte maturation and development. Biol. Reprod. 81, 1064–72.
Marei, W.F., Wathes, D.C. & Fouladi-Nashta, A.A. (2010). Impact of linoleic acid on bovine oocyte maturation and embryo development. Reproduction 139, 979–88.
Marei, W.F., Wathes, D.C. & Fouladi-Nashta, A.A. (2012). Differential effects of linoleic and alpha-linolenic fatty acids on spatial and temporal mitochondrial distribution and activity in bovine oocytes. Reprod. Fertil. Dev. 24, 679–90.
Mattos, R., Staples, C.R. & Thatcher, W.W. (2000). Effects of dietary fatty acids on reproduction in ruminants. Rev. Reprod. 5, 3845.
McKeegan, P.J. & Sturmey, R.G. (2012). The role of fatty acids in oocyte and early embryo development. Reprod. Fertil. Dev. 24, 5967.
Mohammadi-Sangcheshmeh, A., Soleimani, M., Deldar, H., Salehi, M., Soudi, S., Hashemi, S.M., Schellander, K. & Hoelker, M. (2011). Prediction of oocyte developmental competence in ovine using glucose-6-phosphate dehydrogenase (G6PDH) activity determined at retrieval time. J. Assist. Reprod. Genet. 29, 153–8.
O’Fallon, J.V., Busboom, J.R., Nelson, M.L. & Gaskins, C.T. (2007). A direct method for fatty acid methyl ester synthesis: application to wet meat tissues, oils, and feedstuffs. J. Anim. Sci. 85, 1511–21.
Ruvolo, P.P., Deng, X. & May, W.S. (2001). Phosphorylation of Bcl2 and regulation of apoptosis. Leukemia 15, 515–22.
Santos, J.E., Bilby, T.R., Thatcher, W.W., Staples, C.R. & Silvestre, F.T. (2008). Long chain fatty acids of diet as factors influencing reproduction in cattle. Reprod. Domest. Anim. 43 (Suppl. 2), 2330.
Song, J.H., Fujimoto, K. & Miyazawa, T. (2000). Polyunsaturated (n-3) fatty acids susceptible to peroxidation are increased in plasma and tissue lipids of rats fed docosahexaenoic acid-containing oils. J. Nutr. 130, 3028–33.
Staples, C.R., Burke, J.M. & Thatcher, W.W. (1998). Influence of supplemental fats on reproductive tissues and performance of lactating cows. J. Dairy Sci. 81, 856–71.
Sturmey, R.G., Reis, A., Leese, H.J. & McEvoy, T.G. (2009). Role of fatty acids in energy provision during oocyte maturation and early embryo development. Reprod. Domest. Anim. 44 (Suppl. 3), 50–8.
Thompson, J.G., Gardner, D.K., Pugh, P.A., McMillan, W.H. & Tervit, H.R. (1995). Lamb birth weight is affected by culture system utilized during in vitro pre-elongation development of ovine embryos. Biol. Reprod. 53, 1385–91.
Torres, A., Batista, M., Diniz, P., Silva, E., Mateus, L. & Lopes-da-Costa, L. (2014). Effects of oocyte donor age and embryonic stage of development on transcription of genes coding for enzymes of the prostaglandins and progesterone synthesis pathways in bovine in vitro produced embryos. Zygote 2014, 111 DOI: 10.1017/S0967199414000446.
Turathum, B., Saikhun, K., Sangsuwan, P. & Kitiyanant, Y. (2010). Effects of vitrification on nuclear maturation, ultrastructural changes and gene expression of canine oocytes. Reprod. Biol. Endocrinol. 8, 70.
Ulloa, S.M., Heinzmann, J., Herrmann, D., Timmermann, B., Baulain, U., Grossfeld, R., Diederich, M., Lucas-Hahn, A. & Niemann, H. (2015). Effects of different oocyte retrieval and in vitro maturation systems on bovine embryo development and quality. Zygote 23, 367–77.
Van Hoeck, V., Leroy, J.L., Arias Alvarez, M., Rizos, D., Gutierrez-Adan, A., Schnorbusch, K., Bols, P.E., Leese, H.J. & Sturmey, R.G. (2013). Oocyte developmental failure in response to elevated nonesterified fatty acid concentrations: mechanistic insights. Reproduction 145, 3344.
Veshkini, A., Asadi, H., Khadem, A.A., Mohammadi-Sangcheshmeh, A., Khazabi, S., Aminafshar, M., Deldar, H., Soleimani, M. & Cinar, M.U. (2015). Effect of Linolenic acid during in vitro maturation of ovine oocytes: embryonic developmental potential and mRNA abundances of genes involved in apoptosis. J. Assist. Reprod. Genet. 32, 653–59.
Wan, P.C., Hao, Z.D., Zhou, P., Wu, Y., Yang, L., Cui, M.S., Liu, S.R. & Zeng, S.M. (2009). Effects of SOF and CR1 media on developmental competence and cell apoptosis of ovine in vitro fertilization embryos. Anim. Reprod. Sci. 114, 279–88.
Wang, Q. & Sun, Q.Y. (2007). Evaluation of oocyte quality: morphological, cellular and molecular predictors. Reprod. Fertil. Dev. 19, 112.
Wonnacott, K.E., Kwong, W.Y., Hughes, J., Salter, A.M., Lea, R.G., Garnsworthy, P.C. & Sinclair, K.D. (2009). Dietary omega-3 and -6 polyunsaturated fatty acids affect the composition and development of sheep granulosa cells, oocytes and embryos. Reproduction 139, 5769.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

  • ISSN: 0967-1994
  • EISSN: 1469-8730
  • URL: /core/journals/zygote
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed