Hostname: page-component-8448b6f56d-m8qmq Total loading time: 0 Render date: 2024-04-19T15:49:48.226Z Has data issue: false hasContentIssue false

Is it possible to alter the embryo lipid accumulation with reduction of fetal bovine serum and use of l-carnitine for in vitro maturation of bubaline oocytes?

Published online by Cambridge University Press:  10 January 2020

Marivaldo Rodrigues Figueiró*
Affiliation:
Embrapa Eastern Amazon, Belém, Pará, Brazil São Paulo State University, School of Agrarian and Veterinary Sciences, Department of Preventive Veterinary Medicine and Animal Reproduction, Jaboticabal, São Paulo, Brazil
Joaquim Mansano Garcia
Affiliation:
São Paulo State University, School of Agrarian and Veterinary Sciences, Department of Preventive Veterinary Medicine and Animal Reproduction, Jaboticabal, São Paulo, Brazil
Marina Ragagnin de Lima
Affiliation:
São Paulo State University, School of Agrarian and Veterinary Sciences, Department of Preventive Veterinary Medicine and Animal Reproduction, Jaboticabal, São Paulo, Brazil
Maite del Collado
Affiliation:
Veterinary Medicine Department, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga, São Paulo, Brazil
Naiara Zoccal Saraiva
Affiliation:
Embrapa Dairy Cattle, Juiz de Fora, Minas Gerais, Brazil
*
Author for correspondence: Marivaldo Rodrigues Figueiró. Embrapa Amazônia Oriental, Trav. Dr. Enéas Pinheiro, s/no, Caixa Postal 48, CEP 66095-903, Belém, PA, Brazil. Tel: +55 9132041236. E-mail: marivaldo.figueiro@embrapa.br

Summary

In vitro embryo production (IVEP) is a procedure that can promote genetic improvement in a short time frame. However, the success rates obtained with this biotechnology in water buffaloes are still inconsistent, and can be associated with the high concentration of lipids in the cytoplasm of oocytes and embryos. The objective of this study was to evaluate the effects of reduced concentration of fetal bovine serum (FBS) and/or use of l-carnitine during in vitro maturation (IVM) on the preimplantation development and lipid accumulation in bubaline embryos. In a first experiment, the lowest concentration of FBS in the IVM medium (0%, 2.5%, 5% or 10%) was determined, and the lowest concentration that maintained good embryo development rates was 5%. In a second experiment, the addition of 5 mM of l-carnitine into the maturation medium was evaluated. The blastocysts produced were submitted to lipid evaluation involving staining followed by observation using optical (Oil Red O) and confocal (BODIPY 493/503) microscopy. No difference was observed between the 5% and 10% FBS groups, which were superior to the 0% and 2.5% groups. Furthermore, the performance of the groups treated with 5% and 10% FBS was better than the groups supplemented with l-carnitine. There was no difference regarding embryo lipid accumulation. The results indicated that it is possible to reduce the FBS concentration to 5% in in vitro maturation medium for production of bubaline embryos, and supplementation with 5 mM l-carnitine does not increase embryo production.

Type
Research Article
Copyright
© Cambridge University Press 2020

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abdelrazik, H, Sharma, R, Mahfouz, R and Agarwal, A (2009) l-Carnitine decreases DNA damage and improves the in vitro blastocyst development rate in mouse embryos. Fertil Steril 91, 589–96.CrossRefGoogle ScholarPubMed
Abe, H and Hoshi, H (2003) Evaluation of bovine embryos produced in high performance serum-free media. J Reprod Dev 49, 193202.CrossRefGoogle ScholarPubMed
Ali, A and Sirard, MA (2002) Effect of the absence or presence of various protein supplements on further development of bovine oocyte during in vitro maturation. Biol Reprod 66, 901–5.CrossRefGoogle ScholarPubMed
Boni, R, Sangella, L, Dale, B, Rovello, S, Di Palo, R and Barbieri, V (1992) Maturazione in vitro di oociti bufalini: indagine ultrastrutturale. Acta Med Vet 38, 153–61.Google Scholar
Chankitisakul, V, Somfai, T, Inaba, Y, Techakumphu, M and Nagai, T (2013) Supplementation of maturation medium with l-carnitine improves cryo-tolerance of bovine in vitro matured oocytes. Theriogenology 79, 590–8.CrossRefGoogle ScholarPubMed
Chen, J, Hudson, E, Chi, MM, Chang, AS, Moley, KH, Hardie, DG, Downs, SM (2006) AMPK regulation of mouse oocyte meiotic resumption in vitro. Dev Biol 291, 227–38.CrossRefGoogle ScholarPubMed
Del Collado, M, Saraiva, NZ, Lopes, FL, Cruz, MH, Gaspar, RC, Oliveira, CS, Perecin, F and Garcia, JM (2014) Efeitos da redução ou substituição do soro fetal bovino por outros compostos na maturação in vitro de oócitos bovinos. Pesq Vet Bras 34, 689–94.CrossRefGoogle Scholar
Del Collado, M, Saraiva, NZ, Lopes, FL, Gaspar, RC, Padilha, LCCosta, RR, Rossi, GF, Vantini, R and Garcia, JM (2016) Influence of bovine serum albumin and fetal bovine serum supplementation during in vitro maturation on lipid and mitochondrial behaviour in oocytes and lipid accumulation in bovine embryos. Reprod Fertil Dev 28, 1721–32.CrossRefGoogle Scholar
Downs, SM, Mosey, JL and Klinger, J (2009) Fatty acid oxidation and meiotic resumption in mouse oocytes. Mol Reprod Dev 76, 844–53.CrossRefGoogle ScholarPubMed
Dunning, KR and Robker, LR (2012) Promoting lipid utilization with l-carnitine to improve oocyte quality. Anim Reprod Sci 134, 6975.CrossRefGoogle ScholarPubMed
Dunning, KR, Cashman, K, Russell, DL, Thompson, JG, Norman, RJ and Robker, RL (2010) Beta-oxidation is essential for mouse oocyte developmental competence and early embryo development. Biol Reprod 83, 909–18.CrossRefGoogle ScholarPubMed
Dunning, KR, Akison, LK, Russell, DL, Norman, RJ and Robker, RL (2011) Increased beta-oxidation and improved oocyte developmental competence in response to l-carnitine during ovarian in vitro follicle development in mice. Biol Reprod 85, 548–55.CrossRefGoogle ScholarPubMed
Fathi, M and El-Shahat, KH (2017) l-carnitine enhances oocyte maturation and improves in vitro development of embryos in dromedary camels (Camelus dromedaries). Theriogenology 104, 1822.CrossRefGoogle Scholar
Ferraz, ML, Watanabe, YF, Joaquim, DC, Watanabe, MR, Accorsi, MF, Meirelles, FV, Sá Filho, MF and Baruselli, PS (2005) Produção in vitro de embriões bubalinos – resultados preliminaries. Acta Sci Vet 33, 397.Google Scholar
Ferreira, GC and McKenna, MC (2017) l-Carnitine and acetyl-l-carnitine roles and neuroprotection in developing brain. Neurochem Res 42, 1661–75.CrossRefGoogle ScholarPubMed
Feugang, J M, Camargo-Rodríguez, O and Memili, E (2009) Culture systems for bovine embryos. Livest Sci 121, 141–9.CrossRefGoogle Scholar
Gasparrini, B (2002) In vitro embryo production in buffalo species: state of the art 2002. Theriogenology 57, 237–56.CrossRefGoogle Scholar
Gasparrini, B, Sayoud, H, Neglia, G, Matos, DG, Donnay, I and Zicarelli, L (2003) Glutathione synthesis during in vitro maturation of buffalo (Bubalus bubalis) oocytes: effects of cysteamine on embryo development. Theriogenology 60, 943–52.CrossRefGoogle ScholarPubMed
Gasparrini, B, Boccia, L, Marchandise, J, Di Palo, R, George, F, Donnay, I and Zicarelli, L (2006) Enrichment of in vitro maturation medium for buffalo (Bubalus bubalis) oocytes with thiol compounds: effects of cystine on glutathione synthesis and embryo development. Theriogenology 65, 275–87.CrossRefGoogle ScholarPubMed
Gasparrini, B, De Rosa, A, Attanasio, L, Boccia, L, Di Palo, R, Campanile, G and Zicarelli, L (2008) Influence of the duration of in vitro maturation and gamete co-incubation on the efficiency of in vitro embryo development in Italian Mediterranean buffalo (Bubalus bubalis). Anim Reprod Sci 105, 354–64.CrossRefGoogle Scholar
Ghanem, N, Ha, AN, Fakruzzaman, M, Bang, JI, Lee, SC and Kong, IK (2014) Differential expression of selected candidate genes in bovine embryos produced in vitro and cultured with chemicals modulating lipid metabolism. Theriogenology 82, 238–50.CrossRefGoogle ScholarPubMed
Held-Hoelker, E, Klein, SL, Rings, F, Salilew-Wondim, D, Saeed-Zidane, M, Neuhoff, C, Tesfaye, D, Schellander, K and Hoelker, M (2017) Cryosurvival of in vitro produced bovine embryos supplemented with l-carnitine and concurrent reduction of fatty acids. Theriogenology 96, 145–52.CrossRefGoogle ScholarPubMed
Khanmohammadi, N, Movahedin, M, Safari, M, Sameni, HMBehpour Yousefi, B, Jafari, B, Zarbakhsh, S and Zarbakhsh, S (2016) Effect of l-carnitine on in vitro developmental rate, the zona pellucida and hatching of blastocysts and their cell numbers in mouse embryos. Int J Reprod BioMed 14, 649–56.Google ScholarPubMed
Knitlova, D, Hulinska, P, Jeseta, M, Hanzalova, K, Kempisty, B and Machatkova, M (2017) Supplementation of l-carnitine during in vitro maturation improves embryo development from less competent bovine oocytes. Theriogenology 102, 1622.CrossRefGoogle ScholarPubMed
Leivas, FG, Brum, DS, Fialho, SS, Saliba, WP, Alvim, MTT, Bernardic, M L, Rubin, MIB and Silva, CAM (2011) Fetal calf serum enhances in vitro production of Bos taurus indicus embryos. Theriogenology 75, 429–33.CrossRefGoogle ScholarPubMed
Lim, KT, Jang, G, Ko, KH, Lee, WW, Park, HJ, Kim, JJ, Lee, SH, Hwang, WS, Lee, BC and Kang, SK (2007) Improved in vitro bovine embryo development and increased efficiency in producing viable calves using defined media. Theriogenology 67, 293302.CrossRefGoogle ScholarPubMed
Lim, KT, Jang, G, Ko, KH, Lee, WW, Park, HJ, Kim, JJ, Kang, SK and Lee, BC (2008) Improved cryopreservation of bovine preimplantation embryos cultured in chemically defined medium. Anim Reprod Sci 103, 239–48.CrossRefGoogle ScholarPubMed
Lima, MR (2015) Efeitos de reguladores do metabolismo lipídico no desenvolvimento in vitro de embriões bovinos e na sobrevivência à vitrificação Tese (Doutorado em Medicina Veterinária – Reprodução Animal) – Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista “Júlio de Mesquita Filho” Jaboticabal.Google Scholar
Liu, J, Head, E, Kuratsune, H, Cotman, C W and Ames, BN (2004) Comparison of the effects of l-carnitine and acetyl-l-carnitine on carnitine levels ambulatory activity and oxidative stress biomarkers in the brain of old rats. Ann NY Acad Sci 1033, 117–31.CrossRefGoogle ScholarPubMed
Longobardi, V, Salzano, A, Campanile, G, Marrone, R, Palumbo, F, Vitiello, M, Zullo, G, Gasparrini, B (2017) Carnitine supplementation decreases capacitation-like changes of frozen–thawed buffalo spermatozoa. Theriogenology 88, 236–43.CrossRefGoogle ScholarPubMed
Malaguarnera, M (2013) Acetyl-l-carnitine in hepatic encephalopathy. Metab Brain Dis 28, 193–9.CrossRefGoogle ScholarPubMed
Manjunatha, BM, Ravindra, JP, Gupta, PS, Devaraj, M AND Nandi, S (2009) Effect of breeding season on in vivo oocyte recovery and embryo production in non-descriptive Indian river buffaloes (Bubalus bubalis). Anim Reprod Sci 111, 376–83.CrossRefGoogle Scholar
Mingoti, GZ, Castro, VSDC, Méo, S C, Barreto, LSS and Garcia, JM (2011) The effects of macromolecular and serum supplements and oxygen tension during bovine in vitro procedures on kinetics of oocyte maturation and embryo development. In Vitro Cel Dev Biol 47, 361–7.CrossRefGoogle ScholarPubMed
Mishra, A, Reddy, IJ, Gupta, PSP and Mondal, S (2016) L-carnitine mediated reduction in oxidative stress and alteration in transcript level of antioxidant enzymes in sheep embryos produced in vitro. Reprod Dom Anim 51, 311–21.CrossRefGoogle ScholarPubMed
Moawad, AR, Xu, B, Tan, SL and Taketo, T (2014) L-carnitine supplementation during vitrification of mouse germinal vesicle stage-oocytes and their subsequent in vitro maturation improves meiotic spindle configuration and mitochondrial distribution in metaphase II oocytes. Hum Reprod 29, 2256–68.CrossRefGoogle ScholarPubMed
Phongnimitr, T, Liang, Y, Srirattana, K, Panyawai, K, Sripunya, N, Treetampinich, C and Parnpai, R (2013a) Effect of l-carnitine on maturation, cryo-tolerance and embryo developmental competence of bovine oocytes. Anim Sci J 84, 719–25.CrossRefGoogle ScholarPubMed
Phongnimitr, T, Liang, Y, Srirattana, K, Panyawai, K, Sripunya, N, Treetampinich, C and Parnpai, R (2013b) Effects of l-carnitine supplemented in maturation medium on the maturation rate of swamp buffalo oocytes. Buf Bull 32, 613–6.Google Scholar
Reader, KL, Cox, NR, Stanton, JAL and Juengel, JL (2015) Effects of acetyl-l-carnitine on lamb oocyte blastocyst rate, ultrastructure, and mitochondrial DNA copy number. Theriogenology 83, 1484–92.CrossRefGoogle ScholarPubMed
Rizos, D, Gutierrez-Adan, A, Perez-Garnelo, S, De La Fuente, J, Boland, M P and Lonergan, P (2003) Bovine embryo culture in the presence or absence of serum: implications for blastocyst development cryotolerance and messenger RNA expression. Biol Reprod 68, 236–43.CrossRefGoogle ScholarPubMed
Romek, M, Gajda, B, Krzysztofowicz, E, Kepczynski, M, Smorag, Z (2010) New technique to quantify the lipid composition of lipid droplets in porcine oocytes and pre-implantation embryos using Nile red fluorescent probe. Theriogenology 75, 4254.CrossRefGoogle ScholarPubMed
Sá Filho, MF, Carvalho, NAT, Gimenes, LU, Torres-Júnior, JR, Nasser, LF, Tonhati, H, Garcia, JM, Gasparrini, B, Zicarelli, L and Baruselli, PS (2009) Effect of recombinant bovine somatotropin (bST) on follicular population and on in vitro buffalo embryo production. Anim Reprod Sci 113, 51–9.CrossRefGoogle ScholarPubMed
Sayirkava, H, Misirlioglu, M, Kaya, A, First, NL, Parrish, JJ and Memili, E (2007) Developmental potential of bovine oocytes cultured in different maturation and culture conditions. Anim Reprod Sci 1001, 225–40.Google Scholar
Somfai, T, Kaneda, M, Akagi, S, Watanabe, S, Haraguchi, S, Mizutani, EDang-Nguyen, TQGeshi, MKikuchi, K and Nagai, T (2011) Enhancement of lipid metabolism with l-carnitine during in vitro maturation improves nuclear maturation and cleavage ability of follicular porcine oocytes. Reprod Fertil Dev 23, 912–20.CrossRefGoogle ScholarPubMed
Spricigo, JF, Morato, R, Arcarons, N, Yeste, M, Dode, MA, Lopez-Bejar, M and Mogas, T (2017) Assessment of the effect of adding l-carnitine and/or resveratrol to maturation medium before vitrification on in vitro-matured calf oocytes. Theriogenology 89, 4757.CrossRefGoogle ScholarPubMed
Wrenzycki, C, Herrmann, D and Niemann, H (2007) Messenger RNA in oocytes and embryos in relation to embryo vitality. Theriogenology 68, 7783.CrossRefGoogle Scholar
Xu, HY, Yang, XG, Lu, SS, Liang, XW, Lu, YQ, Zhang, M and Lu, KH (2018) Treatment with acetyl-l-carnitine during in vitro maturation of buffalo oocytes improves oocyte quality and subsequent embryonic development. Theriogenology 118, 80–9.CrossRefGoogle ScholarPubMed
Xu, HY, Geng, SS, Li, TT, Fu, Q, Lu, SS, Liang, XW, Lu, YQ, Zhang, M, Yang, XG and Lu, KH (2019) Maturation of buffalo oocytes in vitro with acetyl-l-carnitine improves cryotolerance due to changes in mitochondrial function and the membrane lipid profile. Reprod Fertil Dev 31, 386–94.CrossRefGoogle ScholarPubMed
Yamada, T, Imai, H and Yamada, M (2006) Beneficial effects of acetyl-l-carnitine treatment during IVM on post-fertilization development of bovine oocytes in vitro. Reprod Fertil Dev 18, 280–1.CrossRefGoogle Scholar
Yang, X, Dunning, KR, Wu, LL-Y, Hickey, TE, Norman, RJ, Russel, DL, Liang, X and Robker, RL (2010) Identification of perilipin-2 as a lipid droplet protein regulated in oocytes during maturation. Reprod Fertil Dev 22, 1262–71.CrossRefGoogle ScholarPubMed
Zare, Z, Farahani, RM, Salehi, M, Piryaei, A, Novin, MGFathabadi, FFMohammadi, M and Dehghani-Mohammadabadi, M (2015) Effect of l-carnitine supplementation on maturation and early embryo development of immature mouse oocytes selected by brilliant cresyl blue staining. J Assist Reprod Genet 32, 635–43.CrossRefGoogle Scholar
Zhang, R, Zhang, H, Zhang, Z, Wang, T, Niu, J, Cui, D and Xu, S (2012) Neuroprotective effects of pre-treatment with l-carnitine and acetyl-l-carnitine on ischemic injury in vivo and in vitro. Int J Mol Sci 13, 2078–90.CrossRefGoogle ScholarPubMed