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

  • E. G. Prates (a1) (a2), C. C. Marques (a1), M. C. Baptista (a1), M. I. Vasques (a1), N. Carolino (a1) (a3), A. E. M. Horta (a1), R. Charneca (a2), J. T. Nunes (a2) and R. M. Pereira (a1) (a3)...
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.

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      Fat area and lipid droplet morphology of porcine oocytes during in vitro maturation with trans-10, cis-12 conjugated linoleic acid and forskolin
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E-mail: rosalnp@gmail.com
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B Ambruosi , GM Lacalandra , AI Iorga , T De Santis , S Mugnier , R Matarrese , G Goudet , ME Dell'Aquila 2009. Cytoplasmic lipid droplets and mitochondrial distribution in equine oocytes: implications on oocyte maturation, fertilization and developmental competence after ICSI. Theriogenology 71, 10931104.

JR Dobrinsky , H Nagashima , VG Pursel , CR Long , LA Johnson 1999. Cryopreservation of swine embryos with reduced lipid content. Theriogenology 51, 164.

X Fu , G Wu , G Li , Y Hou , G Zhou , Y Wang , S Zhu 2011. Positive effects of Forskolin (stimulator of lipolysis) treatment on cryosurvival of in vitro matured porcine oocytes. Theriogenology 75, 268275.

T Fujihira , M Kinoshita , M Sasaki , M Ohnishi , H Ishikawa , S Ohsumi , Y Fukui 2004. Comparative studies on lipid analysis and ultrastructure in porcine and southern minke whale (Balaenoptera bonaerensis) oocytes. Journal of Reproduction and Development 50, 525532.

AJ Gardner , JP Evans 2006. Mammalian membrane block to polyspermy: new insights into how mammalian eggs prevent fertilisation by multiple sperm. Reproduction, Fertility and Development 18, 5361.

G Genicot , JLMR Leroy , A Van Soom , I Donnay 2005. The use of a fluorescent dye, Nile red, to evaluate the lipid content of single mammalian oocytes. Theriogenology 63, 11811194.

LDC Gesink , RF Maclehose , MP Longnecker 2007. Obesity and time to pregnancy. Human Reproduction 22, 414420.

MA Gil , LR Abeydeera , BN Day , JM Vazquez , J Roca , EA Martinez 2003. Effect of the volume of medium and number of oocytes during in vitro fertilization on embryo development in pigs. Theriogenology 60, 767776.

V Isachenko , E Isachenko , G Rahimi , P Mallmann , A Krivokhrchenko , F Nawroth 2003. Ultra-structure of intracellular lipid vesicles of porcine GV-oocytes after in vitro fertilization and parthenogenetic activation. Anatomia, Histologia, Embryologia 32, 126128.

MF Laforest , É Pouliot , L Guéguen , FJ Richard 2005. Fundamental significance of specific phosphodiesterases in the control of spontaneous meiotic resumption in porcine oocytes. Molecular Reproduction and Development 70, 361372.

M Lapa , CC Marques , SP Alves , MI Vasques , MC Baptista , I Carvalhais , M Silva Pereira , AEM Horta , RJB Bessa , RM Pereira 2011. Effect of trans-10, cis-12 conjugated linoleic acid on bovine oocyte competence and fatty acid composition. Reproduction Domestic Animals 46, 904910.

TG McEvoy , GD Coull , PJ Broadbent , JSM Hutchinnson , BK Speake 2000. Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida. Journal of Reproduction and Fertility 118, 163170.

H Men , Y Agca , LK Riley , JK Critser 2006. Improved survival of vitrified porcine embryos after partial delipation through chemically stimulated lipolysis and inhibition of apoptosis. Theriogenology 66, 20082016.

RM Pereira , CC Marques 2008. Animal oocyte and embryo cryopreservation. Cell Tissue Banking 9, 267277.

RM Pereira , MC Baptista , MI Vasques , AEM Horta , PV Portugal , RJB Bessa , JC Silva , MS Pereira , CC Marques 2007. Cryosurvival of bovine blastocysts is enhanced by culture with trans-10 cis-12 conjugated linoleic acid (10t,12c CLA). Animal Reproduction Science 98, 293301.

RM Pereira , I Carvalhais , J Pimenta , MC Baptista , MI Vasques , AEM Horta , IC Santos , MR Marques , A Reis , MS Pereira , CC Marques 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, 322332.

C Racowsky 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, 921.

RC Silva , SN Báo , JL Jivago , CM Lucci 2011. Ultrastructural characterization of porcine oocytes and adjacent follicular cells during follicle development: lipid component evolution. Theriogenology 76, 16471657.

RE Thomas , JG Thompson , DT Armstrong , RB Gilchrist 2004. Effect of specific phosphodiesterase isoenzyme inhibitors during in vitro maturation of bovine oocytes on meiotic and developmental capacity. Biology of Reproduction 71, 11421149.

TC Walther , RV Farese 2009. The life of lipid droplets. Biochimica et Biophysica Acta 1791, 459466.

H Wang , N Rodriguez-Osorio , JM Jean Feugang , S Jung , K Garrison , C Wolgemuth , L Greer , M Crenshaw , E Memili 2009. Effects of culture media and inhibitors on biology of porcine early embryonic development in vitro. Livestock Science 121, 102107.

GL Xia , K Kikuchi , J Noguchi , Y Izaike 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, 18071815.

JK Zehmer , Y Huang , G Peng , J Pu , RGW Anderson , P Liu 2009. A role for lipid droplets in inter-membrane lipid traffic. Proteomics 9, 914921.

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animal
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