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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

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: rosalnp@gmail.com
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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

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