Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-26T13:56:02.667Z Has data issue: false hasContentIssue false
  • English
  • Français

Fat area and lipid droplet morphology of porcine oocytes during in vitro maturation with trans-10, cis-12 conjugated linoleic acid and forskolin

Published online by Cambridge University Press:  26 October 2012

E. G. Prates ,
C. C. Marques ,
M. C. Baptista ,
M. I. Vasques ,
N. Carolino ,
A. E. M. Horta ,
R. Charneca ,
J. T. Nunes  and
R. M. Pereira
E. G. Prates
Affiliation:
L-INIAV Santarém, Quinta da Fonte Boa, 00-048 Vale de Santarém, Portugal Universidade de Évora – Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), 7002-554 Évora, Portugal
C. C. Marques
Affiliation:
L-INIAV Santarém, Quinta da Fonte Boa, 00-048 Vale de Santarém, Portugal
M. C. Baptista
Affiliation:
L-INIAV Santarém, Quinta da Fonte Boa, 00-048 Vale de Santarém, Portugal
M. I. Vasques
Affiliation:
L-INIAV Santarém, Quinta da Fonte Boa, 00-048 Vale de Santarém, Portugal
N. Carolino
Affiliation:
L-INIAV Santarém, Quinta da Fonte Boa, 00-048 Vale de Santarém, Portugal Escola Universitária Vasco da Gama, Mosteiro de S. Jorge de Milréu, 3040-714 Coimbra, Portugal
A. E. M. Horta
Affiliation:
L-INIAV Santarém, Quinta da Fonte Boa, 00-048 Vale de Santarém, Portugal
R. Charneca
Affiliation:
Universidade de Évora – Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), 7002-554 Évora, Portugal
J. T. Nunes
Affiliation:
Universidade de Évora – Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), 7002-554 Évora, Portugal
R. M. Pereira*
Affiliation:
L-INIAV Santarém, Quinta da Fonte Boa, 00-048 Vale de Santarém, Portugal Escola Universitária Vasco da Gama, Mosteiro de S. Jorge de Milréu, 3040-714 Coimbra, Portugal
*
†E-mail: [email protected]
Get access

Abstract

Lipid droplets (LD) in porcine oocytes form a dark mass reaching almost all cytoplasm. Herein we investigated changes in fat areas, cytoplasmic tone and LD morphology during in vitro maturation (IVM) of porcine oocytes cultured with 100 μM trans-10, cis-12 conjugated linoleic acid (t10,c12 CLA) or 10 μM forskolin at different time periods. Four groups were constituted: control, excipient, t10,c12 CLA and forskolin, with drugs being supplemented during 44 to 48 h and the initial 22 to 24 h in Experiments 1 and 2, respectively. In Experiment 3, forskolin was supplemented for the first 2 h. Matured oocytes were inseminated with frozen-thawed boar semen and cleavage rate recorded. Before and during IVM, samples of oocytes were evaluated for LD, total and fat areas and fat gray value or for meiotic progression. Results showed that forskolin supplementation during 44 to 48 h or 22 to 24 h inhibits oocyte maturation (exp. 1: forskolin = 5.1 ± 8.0%, control = 72.6 ± 5.0%; exp. 2: forskolin = 24.3 ± 7.4%, control = 71.6 ± 5.6%) and cleavage (exp. 1: forskolin = 0.0 ± 0.0%, control = 55.4 ± 4.1%; exp. 2: forskolin = 8.3 ± 3.3%, control = 54.5 ± 3.0%). Forskolin also reduced oocyte and fat areas. In Experiment 3, forskolin negative effect on oocyte maturation and cleavage disappeared, although minor (P ⩽ 0.03) LD and oocyte fat areas were identified at 22 to 24 h of IVM. Oocytes supplemented with t10,c12 CLA during 44 to 48 h presented a lighter (P ⩽ 0.04) colour tone cytoplasm than those of control and forskolin. In conclusion, t10,c12 CLA and forskolin were capable of modifying the distribution and morphology of cytoplasmic LD during porcine oocyte maturation, thus reducing its lipid content in a time-dependent manner.

Keywords

lipid dropletsporcine oocyte maturationlipid contentconjugated linoleic acidforskolin
Type
Physiology and functional biology of systems
Information
animal , Volume 7 , Issue 4 , April 2013 , pp. 602 - 609
Copyright
Copyright © The Animal Consortium 2012

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

Ambruosi, B, Lacalandra, GM, Iorga, AI, De Santis, T, Mugnier, S, Matarrese, R, Goudet, G, Dell'Aquila, ME 2009. Cytoplasmic lipid droplets and mitochondrial distribution in equine oocytes: implications on oocyte maturation, fertilization and developmental competence after ICSI. Theriogenology 71, 1093–1104.Google Scholar
Corino, C, Pastorelli, G, Douard, V, Rossi, R, Musella, M, Mourot, J 2006. L'acid linoléique conjugue en nutrition porcine. INRA Productions Animales 19, 39–46.Google Scholar
Dobrinsky, JR, Nagashima, H, Pursel, VG, Long, CR, Johnson, LA 1999. Cryopreservation of swine embryos with reduced lipid content. Theriogenology 51, 164.Google Scholar
Fu, X, Wu, G, Li, G, Hou, Y, Zhou, G, Wang, Y, Zhu, S 2011. Positive effects of Forskolin (stimulator of lipolysis) treatment on cryosurvival of in vitro matured porcine oocytes. Theriogenology 75, 268–275.Google Scholar
Fujihira, T, Kinoshita, M, Sasaki, M, Ohnishi, M, Ishikawa, H, Ohsumi, S, Fukui, Y 2004. Comparative studies on lipid analysis and ultrastructure in porcine and southern minke whale (Balaenoptera bonaerensis) oocytes. Journal of Reproduction and Development 50, 525–532.Google Scholar
Gardner, AJ, Evans, JP 2006. Mammalian membrane block to polyspermy: new insights into how mammalian eggs prevent fertilisation by multiple sperm. Reproduction, Fertility and Development 18, 53–61.Google Scholar
Genicot, G, Leroy, JLMR, Van Soom, A, Donnay, I 2005. The use of a fluorescent dye, Nile red, to evaluate the lipid content of single mammalian oocytes. Theriogenology 63, 1181–1194.Google Scholar
Gesink, LDC, Maclehose, RF, Longnecker, MP 2007. Obesity and time to pregnancy. Human Reproduction 22, 414–420.Google Scholar
Gil, MA, Abeydeera, LR, Day, BN, Vazquez, JM, Roca, J, Martinez, EA 2003. Effect of the volume of medium and number of oocytes during in vitro fertilization on embryo development in pigs. Theriogenology 60, 767–776.Google Scholar
Isachenko, V, Isachenko, E, Rahimi, G, Mallmann, P, Krivokhrchenko, A, Nawroth, F 2003. Ultra-structure of intracellular lipid vesicles of porcine GV-oocytes after in vitro fertilization and parthenogenetic activation. Anatomia, Histologia, Embryologia 32, 126–128.Google Scholar
Kikuchi, K, Ekwall, H, Tienthai, P, Kawai, Y, Noguchi, J, Kaneko, H, Rodriguez-Martinez, H 2002. Morphological features of lipid droplet transition during porcine oocyte fertilisation and early embryonic development to blastocyst in vivo and in vitro. Zygote 10, 355–366.Google Scholar
José, AAFBV, Gama, MAS, Lanna, DDP 2008. Effects of trans-10, cis-12 conjugated linoleic acid on gene expression and lipid metabolism of adipose tissue of growing pigs. Genetics and Molecular Research 78, 284–294.Google Scholar
Laforest, MF, Pouliot, É, Guéguen, L, Richard, FJ 2005. Fundamental significance of specific phosphodiesterases in the control of spontaneous meiotic resumption in porcine oocytes. Molecular Reproduction and Development 70, 361–372.Google Scholar
Lapa, M, Marques, CC, Alves, SP, Vasques, MI, Baptista, MC, Carvalhais, I, Silva Pereira, M, Horta, AEM, Bessa, RJB, Pereira, RM 2011. Effect of trans-10, cis-12 conjugated linoleic acid on bovine oocyte competence and fatty acid composition. Reproduction Domestic Animals 46, 904–910.Google Scholar
McEvoy, TG, Coull, GD, Broadbent, PJ, Hutchinnson, JSM, Speake, BK 2000. Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida. Journal of Reproduction and Fertility 118, 163–170.Google Scholar
Men, H, Agca, Y, Riley, LK, Critser, JK 2006. Improved survival of vitrified porcine embryos after partial delipation through chemically stimulated lipolysis and inhibition of apoptosis. Theriogenology 66, 2008–2016.Google Scholar
Pereira, RM, Marques, CC 2008. Animal oocyte and embryo cryopreservation. Cell Tissue Banking 9, 267–277.Google Scholar
Pereira, RM, Baptista, MC, Vasques, MI, Horta, AEM, Portugal, PV, Bessa, RJB, Silva, JC, Pereira, MS, Marques, CC 2007. Cryosurvival of bovine blastocysts is enhanced by culture with trans-10 cis-12 conjugated linoleic acid (10t,12c CLA). Animal Reproduction Science 98, 293–301.CrossRefGoogle Scholar
Pereira, RM, Carvalhais, I, Pimenta, J, Baptista, MC, Vasques, MI, Horta, AEM, Santos, IC, Marques, MR, Reis, A, Pereira, MS, Marques, CC 2008. Biopsied and vitrified bovine embryos viability is improved by trans-10 cis-12 conjugated linoleic acid supplementation during in vitro embryo culture. Animal Reproduction Science 106, 322–332.Google Scholar
Petters, RM, Wells, KD 1993. Culture of pig embryos. Journal of Reproduction and Fertility 48, 61–73.Google Scholar
Racowsky, C 1985. Effect of forskolin on maintenance of meiotic arrest and stimulation of cumulus expansion, progesterone and cyclic AMP production by pig oocyte-cumulus complexes. Journal of Reproduction and Fertility 74, 9–21.Google Scholar
Silva, RC, Báo, SN, Jivago, JL, Lucci, CM 2011. Ultrastructural characterization of porcine oocytes and adjacent follicular cells during follicle development: lipid component evolution. Theriogenology 76, 1647–1657.Google Scholar
Thomas, RE, Thompson, JG, Armstrong, DT, Gilchrist, RB 2004. Effect of specific phosphodiesterase isoenzyme inhibitors during in vitro maturation of bovine oocytes on meiotic and developmental capacity. Biology of Reproduction 71, 1142–1149.Google Scholar
Walther, TC, Farese, RV 2009. The life of lipid droplets. Biochimica et Biophysica Acta 1791, 459–466.Google Scholar
Wang, H, Rodriguez-Osorio, N, Jean Feugang, JM, Jung, S, Garrison, K, Wolgemuth, C, Greer, L, Crenshaw, M, Memili, E 2009. Effects of culture media and inhibitors on biology of porcine early embryonic development in vitro. Livestock Science 121, 102–107.Google Scholar
Xia, GL, Kikuchi, K, Noguchi, J, Izaike, Y 2000. Short time priming of cumulus–oocyte complexes with FSH and forskolin in the presence of hypoxanthine stimulates cumulus cells to secrete a meiosis-activating substance. Theriogenology 53, 1807–1815.Google Scholar
Zehmer, JK, Huang, Y, Peng, G, Pu, J, Anderson, RGW, Liu, P 2009. A role for lipid droplets in inter-membrane lipid traffic. Proteomics 9, 914–921.Google Scholar