Dimethylsulfoxide and conjugated linoleic acids affect bovine embryo development in vitro

Abstract

Conjugated linoleic acids (CLA) are employed to overcome the bovine periparturitional negative energy balance. Especially of interest are trans10,cis12 -linoleic acid (t10c12-CLA) and cis9,trans11-linoleic acid (c9t11-CLA). Their impact on embryonic development, though, is not clear. Here, effects of both above-mentioned CLA on bovine in vitro-produced embryos were assessed. Zygotes (n=2098) were allocated to one of seven groups: cultured with 50 or 100µM of either c9t11-CLA or t10c12-CLA, with 14 or 28mM DMSO or without supplement (control). Messenger RNA analysis of target gene transcripts (IGF1R, IGFBP2, IGFBP4, CPT2, ACAA1, ACAA2, FASN, SCD) via RT-qPCR was performed in single blastocysts. Cleavage rates did not differ, whereas development rates were decreased in both t10c12-supplemented groups in comparison to the unsupplemented group (31.7% ±2.2 control vs 20.2% ±2.0 50µM t10c12 vs 21.0% ±2.8 100µM t10c12). Compared with the unsupplemented group, SCD was expressed at a lower level in embryos cultured with 50µM c9t11-CLA. The relative amount of several transcripts was increased in embryos cultured with 14mM DMSO in comparison to those that developed in the presence of 50µM t10c12-CLA (IGFBP2, ACAA1, CPT2, FASN, SCD) or 50µM c9t11-CLA (IGF1R, IGFBP2, ACAA1, CPT2, FASN, SCD). The molecular analyses show that CLA influence embryonic fat metabolism.

Dietary Fat Supplementation and the Consequences for Oocyte and Embryo Quality: Hype or Significant Benefit for Dairy Cow Reproduction?

“…), while conjugated linoleic acids decrease embryo development rate and also suppress expression of stearoyl‐CoA desaturase‐1, the enzyme that converts stearic acid to oleic acid (Stinshoff et al. ). A very recent study conducted in Brazil could not show any positive effects of feeding extra linoleic acid to Nellore heifers on embryo production.…”

Lipid content and cryotolerance of in vitro-produced bovine embryos treated with forskolin before vitrification

“…Milimolar stock concentrations of forskolin were stored at 20 o C dissolved in anydrous dimethylsulphoxide (DMSO) solutions and were freshly diluted each day of the experiment to obtain a final concentration of 10mM, 5mM and 2.5mM. The maximum final concentration of DMSO in the cultures was 0.08% (according to previous results from Stinshoff et al (2014), the presence of 0.2% DMSO in the culture medium exerted no toxic effects on the developing embryos compared to control).…”

Effect of delipidant agents during in vitro culture on the development, lipid content, gene expression and cryotolerance of bovine embryos

“…Similarly, a lack of effect on Bx rate when cultured in the presence of L‐carnitine has also been reported (Chankitisakul et al, ; Held‐Hoelker et al, ; Phongnimitr et al, ). As demonstrated for carnitine, the results obtained using CLA have also been controversial, with some studies showing a beneficial effect (Ghanem et al, ; Takahashi et al, ) on the embryo and others not showing a beneficial effect (Batista et al, ; Leão et al, ; Stinshoff, Wilkening, Hanstedt, Bollwein, & Wrenzycki, ).…”