Improving the Effects of Omega-3 Fatty Acid on the In Vitro Maturation of Oocytes

Masume Ghorbani Vahed, Ramazan Khanbabaee, Mehrdad Shariati, Mohammad Amin Edalatmanesh


This research was conducted in order to determine the effects of omega-3 on oocyte in vitro maturation and the level of expression of tribbles (TRIB1, TRIB2 and TRIB3 genes) in cumulus cells. Eight-ten weeks old NMRI mice were super-ovulated using 7.5 IU pregnant mare’s serum gonadotropin (PMSG, Intraperitoneal) and they were killed after 44 hours and their ovaries were removed. The oocytes were used for in vitro maturation and the cumulus-oocyte complexes (COCs) were released. Cumulus cells and oocytes were assigned into control, ethanol-treated and groups exposed to 10 and 100 μg/ml of omega-3. The cells were prepared to assess the maturation stage in order to evaluate the gene expression level. The data were statistically analyzed. Exposing oocytes to low dose (10 μg/ml) and high dose (100 μg/ml) of omega-3 resulted in a reduced rate of GV-stage oocytes, decreased MI-oocytes and increased MII-oocytes. The enhanced maturity of COCs was also detected in response to a high dose of omega-3 (100 μg/ml). Exposure of cumulus cells to omega-3 (10 and 100 μg/ml) induced TRIB2 and inhibited TRIB3 gene expression level; however, TRIB1 gene expression level increased and decreased in response to low (10 μg/ml) and high (100 μg/ml) concentrations of omega-3, respectively. The addition of omega-3 to the environment of oocytes or cumulus cells affected the maturation of oocytes and cumulus cells, which was followed by the differential expression of TRIB genes, suggesting that there was a role of fatty acid metabolism in the differentiation and maturation of cumulus cells.


Cumulus Cells, Maturation, Omega-3, Oocyte, TRIB Genes

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Assidi M, Dieleman SJ, Sirard M-A. 2010. Cumulus cell gene expression following the LH surge in bovine preovulatory follicles: potential early markers of oocyte competence. REPRODUCTION. 140:835–852.

Bauer RC, Yenilmez BO, Rader DJ. 2015. Tribbles-1: a novel regulator of hepatic lipid metabolism in humans. Biochem Soc Trans. 43:1079–1084.

Brisard D, Chesnel F, Elis S, Desmarchais A, Sánchez-Lazo L, Chasles M, Maillard V, Uzbekova S. 2014. Tribbles expression in cumulus cells is related to oocyte maturation and fatty acid metabolism. J Ovarian Res. 7:44.

Cao Y, Lu L, Liang J, Liu M, Li X, Sun R, Zheng Y, Zhang P. 2015. Omega-3 fatty acids and primary and secondary prevention of cardiovascular disease. Cell Biochem Biophys. 72:77–81.

Carro M, Buschiazzo J, Ríos GL, Oresti GM, Alberio RH. 2013. Linoleic acid stimulates neutral lipid accumulation in lipid droplets of maturing bovine oocytes. Theriogenology. 79:687–694.

Cheng Z, Abayasekara DRE, Ward F, Preece DMW, Raheem KA, Wathes DC. 2013. Altering n-3 to n-6 polyunsaturated fatty acid ratios affects prostaglandin production by ovine uterine endometrium. Anim Reprod Sci. 143:38–47.

Chiu YH, Karmon AE, Gaskins AJ, Arvizu M, Williams PL, Souter I, Rueda BR, Hauser R, Chavarro JE. 2018. Serum omega-3 fatty acids and treatment outcomes among women undergoing assisted reproduction. Hum Reprod. 33:156–165.

Dirandeh E, Towhidi A, Ansari Pirsaraei Z, Saberifar T, Akhlaghi A, Rezaei Roodbari A. 2015. The endometrial expression of prostaglandin cascade components in lactating dairy cows fed different polyunsaturated fatty acids. Theriogenology. 83:206–212.

Dugast E, Kiss-Toth E, Soulillou J-P, Brouard S, Ashton-Chess J. 2012. The tribbles-1 protein in humans: roles and functions in health and disease. Curr Mol Med. 13:80–85.

Dunning KR, Cashman K, Russell DL, Thompson JG, Norman RJ, Robker RL. 2010. Beta-oxidation is essential for mouse oocyte developmental competence and early embryo development. Biol Reprod 83:909–918.

Fayezi S, Leroy JLMR, Ghaffari Novin M, Darabi M. 2018. Oleic acid in the modulation of oocyte and preimplantation embryo development. Zygote. 26:1–13.

Gulliver CE, Friend MA, King BJ, Clayton EH. 2012. The role of omega-3 polyunsaturated fatty acids in reproduction of sheep and cattle. Anim Reprod Sci. 131:9–22.

Gumus E, Bulut HE, Kaloglu C. 2010. Cytoskeletal changes in oocytes and early embryos during in vitro fertilization process in mice. Anat Histol Embryol. 39:51–58.

Hernández-Montiel W, Collí-Dula RC, Ramón-Ugalde JP, Martínez-Núñez MA, Zamora-Bustillos R. 2019. RNA-seq transcriptome analysis in ovarian tissue of pelibuey breed to explore the regulation of prolificacy. Genes (Basel). 10:358.

Jeromson S, Gallagher I, Galloway S, Hamilton D. 2015. Omega-3 fatty acids and skeletal muscle health. mar drugs.

Khalil WA, Marei WFA, Khalid M. 2013. Protective effects of antioxidants on linoleic acid–treated bovine oocytes during maturation and subsequent embryo development. Theriogenology. 80:161–168.

Kirkup SE, Cheng Z, Elmes M, Wathes DC, Abayasekara DRE. 2010. Polyunsaturated fatty acids modulate prostaglandin synthesis by ovine amnion cells in vitro. REPRODUCTION. 140:943–951.

Legault M-A, Tardif J-C, Dubé M-P. 2018. Pharmacogenomics of blood lipid regulation. Pharmacogenomics. 19:651–665.

Lirangi M, Meydani M, Zingg JM, Azzi A. 2012. α-tocopheryl-phosphate regulation of gene expression in preadipocytes and adipocytes. BioFactors. 38:450–457.

Lohan F, Keeshan K. 2013. The functionally diverse roles of tribbles. Biochem Soc Trans. 41:1096–1100.

Lussier JG, Diouf MN, Lévesque V, Sirois J, Ndiaye K. 2017. Gene expression profiling of upregulated mRNAs in granulosa cells of bovine ovulatory follicles following stimulation with hCG. Reprod Biol Endocrinol. 15:88.

Mahla AS, Chaudhari RK, Verma AK, Singh AK, Singh SK, Singh G, Sarkar M, Dutta N, Kumar H, Krishnaswamy N. 2017. Effect of dietary supplementation of omega-3 polyunsaturated fatty acid (PUFA) rich fish oil on reproductive performance of the goat (Capra hircus). Theriogenology. 99:79–89.

Maillard V, Desmarchais A, Durcin M, Uzbekova S, Elis S. 2018. Docosahexaenoic acid (DHA) effects on proliferation and steroidogenesis of bovine granulosa cells. Reprod Biol Endocrinol. 16:40.

Marei WF, Wathes DC, Fouladi-Nashta AA. 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.

Meher AP, Joshi AA, Joshi SR. 2013. Preconceptional omega-3 fatty acid supplementation on a micronutrient-deficient diet improves the reproductive cycle in Wistar rats. Reprod Fertil Dev. 25:1085.

Meng H, Shen Y, Shen J, Zhou F, Shen S, Das UN. 2013. Effect of n-3 and n-6 unsaturated fatty acids on prostate cancer (PC-3) and prostate epithelial (RWPE-1) cells in vitro. Lipids Health Dis. 12:160.

Mirabi P, Chaichi MJ, Esmaeilzadeh S, Ali Jorsaraei SG, Bijani A, Ehsani M, hashemi Karooee SF. 2017. The role of fatty acids on ICSI outcomes: a prospective cohort study. Lipids Health Dis. 16:18.

Nehra D, Le HD, Fallon EM, Carlson SJ, Woods D, White YA, Pan AH, Guo L, Rodig SJ, Tilly JL, et al. 2012. Prolonging the female reproductive lifespan and improving egg quality with dietary omega-3 fatty acids. Aging Cell. 11:1046–1054.

Nikoloff N, Pascua AM, Anchordoquy JM, Anchordoquy JP, Sirini MA, Seoane A, Furnus CC. 2017. Effect of eicosapentaenoic acid on bovine cumulus-oocyte complex in vitro. Cell Biol Int [Internet]. 41:505–513. Available from:

Nikseresht M. 2015. The nuclear maturation and embryo development of mice germinal vesicle oocytes with and without cumulus cell after vitrification. J Clin Diagnostic Res. 9:AF01–AF04.

Örd Tiit, Örd D, Adler P, Vilo J, Örd Tõnis. 2015. TRIB3 enhances cell viability during glucose deprivation in HEK293-derived cells by upregulating IGFBP2, a novel nutrient deficiency survival factor. Biochim Biophys Acta - Mol Cell Res. 1853:2492–2505.

Ortiz ME, Bühler MI, Zelarayán LI. 2014. Involvement of PLA 2 , COX and LOX in Rhinella arenarum oocyte maturation. Zygote. 22:440–445.

Oseikria M, Elis S, Maillard V, Corbin E, Uzbekova S. 2016. N-3 polyunsaturated fatty acid DHA during IVM affected oocyte developmental competence in cattle. Theriogenology. 85:1625-1634.e2.

Safdar AHA, Sadeghi AA, Chamani M. 2017. Effects of different fat sources (saturated and unsaturated) on reproductive performance and biological indices of ewes during flushing period. Trop Anim Health Prod. 49:1447–1453.

Shaaker M, Rahimipour A, Nouri M, Khanaki K, Darabi M, Farzadi L, Shahnazi V, Mehdizadeh A. 2012. Fatty acid composition of human follicular fluid phospholipids and fertilization rate in assisted reproductive techniques. Iran Biomed J. 16:162–168.

Sirard M-A. 2011. Follicle environment and quality of in vitro matured oocytes. J Assist Reprod Genet. 28:483–488.

Veshkini A, Khadem AA, Mohammadi-Sangcheshmeh A, Alamouti AA, Soleimani M, Gastal EL. 2016. Linolenic acid improves oocyte developmental competence and decreases apoptosis of in vitro -produced blastocysts in goat. Zygote. 24:537–548.

Virant-Klun I, Knez K, Tomazevic T, Skutella T. 2013. Gene expression profiling of human oocytes developed and matured in vivo or in vitro. Biomed Res Int. 2013:1–20.

Wang L, Jing J, Fu Q, Tang X, Su L, Wu S, Li G, Zhou L. 2015. Association study of genetic variants at newly identified lipid gene TRIB1 with coronary heart disease in Chinese Han population. Lipids Health Dis. 14:46.

Wonnacott K, Kwong W, Hughes J, Salter A, Lea R, Garnsworthy P, Sinclair K. 2020. Dietary omega-3 and -6 polyunsaturated fatty acids affect the composition and development of sheep ranulose cells, oocytes and embryos. Reproduction. 139:57–69.

Wu S, Guo Y, Wu Y, Zhu S, He Z, Chen YQ. 2015. Omega-3 free fatty acids inhibit tamoxifen-induced cell apoptosis. Biochem Biophys Res Commun. 459:292–299.

Zachut M, Dekel I, Lehrer H, Arieli A, Livshitz L, Yakoby S, Moallem U. 2010. Effects of dietary fats differing in n-6:n-3 ratio fed to high-yielding dairy cows on fatty acid composition of ovarian compartments, follicular status, and oocyte quality. J Dairy Sci. 93:529–545.

Zajdel A, Wilczok A, Chodurek E, Gruchlik A, Dzierzewicz Z. 2013. Polyunsaturated fatty acids inhibit melanoma cell growth in vitro. Acta Pol Pharm. 70:365–9.

Zhang W, Wu M, Kim T, Jariwala RH, Garvey WJ, Luo N, Kang M, Ma E, Tian L, Steverson D, et al. 2016. Skeletal Muscle TRIB3 Mediates Glucose Toxicity in Diabetes and High- Fat Diet–Induced Insulin Resistance. Diabetes. 65:2380–2391.

Zhou C-J, Wu S-N, Shen J-P, Wang D-H, Kong X-W, Lu A, Li Y-J, Zhou H-X, Zhao Y-F, Liang C-G. 2016. The beneficial effects of cumulus cells and oocyte-cumulus cell gap junctions depends on oocyte maturation and fertilization methods in mice. PeerJ. 4:e1761.


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