title: Trans-10, cis-12 conjugated linoleic acid reduces neutral lipid content and may affect cryotolerance of in vitro-produced crossbred bovine embryos.
abstract: BACKGROUND
  Due to high neutral lipids accumulation in the cytoplasm, in vitro-produced embryos from Bos primigenius indicus and their crosses are more sensitive to chilling and cryopreservation than those from Bos primigenius taurus. The objective of the present study was to evaluate the effects of trans-10, cis-12 conjugated linoleic acid (CLA) on the development and cryotolerance of crossbred Bos primigenius taurus x Bos primigenius indicus embryos produced in vitro, and cultured in the presence of fetal calf serum. Bovine zygotes (n = 1,692) were randomly assigned to one of the following treatment groups: 1) Control, zygotes cultured in Charles Rosenkrans 2 amino acid (CR2aa) medium (n = 815) or 2) CLA, zygotes cultured in CR2aa medium supplemented with 100 μmol/L of trans-10, cis-12 CLA (n = 877). Embryo development (cleavage and blastocyst rates evaluated at days 3 and 8 of culture, respectively), lipid content at morula stage (day 5) and blastocyst cryotolerance (re-expansion and hatching rates, evaluated 24 and 72 h post-thawing, respectively) were compared between groups. Additionally, selected mRNA transcripts were measured by Real-Time PCR in blastocyst stage.
  
  
  RESULTS
  The CLA treatment had no effect on cleavage and blastocyst rates, or on mRNA levels for genes related to cellular stress and apoptosis. On the other hand, abundance of mRNA for the 1-acylglycerol-3-phosphate 0-acyltransferase-encoding gene (AGPAT), which is involved in triglycerides synthesis, and consequently neutral lipid content, were reduced by CLA treatment. A significant increase was observed in the re-expansion rate of embryos cultured with trans-10, cis-12 CLA when compared to control (56.3 vs. 34.4%, respectively, P = 0.002). However, this difference was not observed in the hatching rate (16.5 vs. 14.0%, respectively, P = 0.62).
  
  
  CONCLUSIONS
  The supplementation with trans-10, cis-12 CLA isomer in culture medium reduced the lipid content of in vitro produced bovine embryos by reducing the gene expression of 1-acylglycerol 3-phosphate 0-acyltransferase (AGPAT) enzyme. However, a possible improvement in embryo cryotolerance in response to CLA, as suggested by increased blastocyst re-expansion rate, was not confirmed by hatching rates.
authors.affiliation: Federal University of Juiz de Fora, Juiz de Fora, MG 36036-900, Brazil ; Embrapa Dairy Cattle Research Center, Juiz de Fora, MG 36038-330, Brazil.
authors.forename: Ribrio Ivan Tavares Pereira
authors.initials: RI
authors.lastname: Batista
authors.affiliation: Federal University of Juiz de Fora, Juiz de Fora, MG 36036-900, Brazil.
authors.forename: Nádia Rezende Barbosa
authors.initials: NR
authors.lastname: Raposo
authors.affiliation: Federal University of Minas Gerais, Belo Horizonte, MG 31270-901, Brazil.
authors.forename: Paulo Henrique Almeida
authors.initials: PH
authors.lastname: Campos-Junior
authors.affiliation: Federal University of Juiz de Fora, Juiz de Fora, MG 36036-900, Brazil.
authors.forename: Michele Munk
authors.initials: MM
authors.lastname: Pereira
authors.affiliation: Embrapa Dairy Cattle Research Center, Juiz de Fora, MG 36038-330, Brazil.
authors.forename: Luiz Sergio Almeida
authors.initials: LS
authors.lastname: Camargo
authors.affiliation: Embrapa Dairy Cattle Research Center, Juiz de Fora, MG 36038-330, Brazil.
authors.forename: Bruno Campos
authors.initials: BC
authors.lastname: Carvalho
authors.affiliation: Embrapa Dairy Cattle Research Center, Juiz de Fora, MG 36038-330, Brazil.
authors.forename: Marco Antonio Sundfeld
authors.initials: MA
authors.lastname: Gama
authors.affiliation: Embrapa Dairy Cattle Research Center, Juiz de Fora, MG 36038-330, Brazil.
authors.forename: João Henrique Moreira
authors.initials: JH
authors.lastname: Viana
keywords: Blastocysts; CLA; Crossbred cattle; Cryopreservation; Lipid content