Abstract
In this study, we evaluated the modulation effect of long-chain Acyl-CoA synthetase during early embryo development. Bovine embryos were cultured in four groups: positive modulation (ACS+) with GW3965 hydrochloride, negative modulation (ACS-) with Triacsin C, association of both modulators (ACS±), and control. Embryo development rates were not altered (P>0.05) by treatments. Embryonic cytoplasmic lipid content increased in ACS+ but reduced in ACS- compared to the control (P < 0.05), whereas the membrane phospholipids profile was not altered by treatments. The total number of blastomeres did not differ (P > 0.05) between groups; however, an increased apoptotic cells percentage was found in ACS- compared to control. Twenty-four hours after warming, ACS+ and control grade I embryos presented the best hatching rates, whereas the ACS+ group equaled the hatching rates between their embryos of grades I, II and III 48 hours after warming. The relative abundance of transcripts for genes associated with lipid metabolism (ACSL3, ACSL6, ACAT1, SCD, and AUH), heatshock (HSP90AA1 and HSF1), oxidative stress (GPX4), and angiogenesis (VEGF), among other important genes for embryo development were affected by at least one of the treatments. The treatments were effective in modulating the level of transcripts for ACSL3 and the cytoplasmic lipid content. The ACS- was not effective in increasing embryonic cryosurvival, whereas ACS+ restored survival rates after vitrification of embryos with low quality, making them equivalent to embryos of excellent quality.
Highlights
Over the past several decades, increased demand for in vitro produced (IVP) bovine embryos has triggered a constant search for improvements of the technique, aiming at the production of embryos of better quality that result in higher pregnancy rates
In view of the satisfactory results obtained with embryos produced in vivo, the reduced cryosurvival of IVP may be directly related to sub-optimal in vitro culture conditions during development, leading to structural and functional modifications that compromise embryo viability
The objective of the present work was to evaluate the effects of the addition of GW3965 hydrochloride and Triacsin C, positive (ACS+) and negative (ACS-) modulators of ACSL, respectively, on development, cryosurvival, lipid accumulation and profile, gene expression, and viability of bovine IVP embryos
Summary
Over the past several decades, increased demand for in vitro produced (IVP) bovine embryos has triggered a constant search for improvements of the technique, aiming at the production of embryos of better quality that result in higher pregnancy rates. The existence of morphological and metabolic differences in IVP embryos has been clearly demonstrated in relation to in vivo-produced ones Among these differences is the higher cytoplasmic lipid content, commonly associated with lower rates of embryonic survival after freezing [1] and deviations in the relative abundance of transcripts of important genes for embryonic development and establishment of gestation [2]. The mechanisms through which IVP embryos accumulate more lipids are not yet fully elucidated; these embryos are known to be less resistant to cryopreservation when compared to those produced in vivo Alternatives such as changes in media culture conditions, like the addition of lipolytic chemicals [3] or the reduction or removal of fetal calf serum [4,5,6] were proposed to decrease the amount of lipid droplets within the embryonic cytoplasm
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