Effect of different culture systems on adipocyte differentiation-related protein (ADRP) in bovine embryos

• Published in: Animal reproduction science Vol. 145; no. 3-4; pp. 105 - 113
• Main Authors: Al Darwich, A., Perreau, C., Tsikis, G., Coudert, E., Touzé, J.L., Briant, E., Beckers, J.F., Mermillod, P., Guignot, F.
• Format: Journal Article Web Resource
• Language: English
• Published: Netherlands Elsevier B.V 01.03.2014; Elsevier [1977-....]; Elsevier
• Subjects: ADRP quantification and localisation; Animal Science and Zoology; Animals; Cattle - embryology; Coculture Techniques - veterinary; Culture environment; Embryo Culture Techniques - veterinary; Embryo, Mammalian - metabolism; Endocrinology; Fertilization in Vitro; Food Animals; Gene Expression Regulation, Developmental - physiology; General Medicine; Life Sciences; Lipid droplet; Membrane Proteins; Membrane Proteins - genetics; Membrane Proteins - metabolism; Médecine vétérinaire & santé animale; Other; Perilipin-2; RNA, Messenger; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sciences du vivant; Sheep; Veterinary medicine & animal health; ADRP quantification and localisation; Culture environment; Lipid droplet; lipid droplet; culture environment; adpr quantification and localisation
• ISSN: 0378-4320; 1873-2232; 1873-2232
• DOI: 10.1016/j.anireprosci.2014.01.010

Bovine embryos cultured in serum-containing media abnormally accumulate lipid droplets, compared to their in vivo counterparts. The objective of this study was to investigate the effect of different culture systems on the mRNA expression and on the quantification and localisation of adipocyte differentiation-related protein (ADRP), a protein associated with lipid accumulation in bovine blastocysts. Two experiments were independently performed for ADRP mRNA expression analysis. In experiment A, blastocysts were produced in modified synthetic oviduct fluid (mSOF)+10% foetal calf serum (FCS), in coculture (bovine oviduct epithelial cells, Boec) and in ewe oviducts, whereas in experiment B, they were produced in mSOF+10μM docosahexaenoic acid (DHA) and in vivo. Control groups were also performed. ADRP mRNA expression was downregulated in the Boec, ewe oviduct and in vivo groups compared to the 10% FCS or DHA groups, respectively. Moreover, the expression of this protein was downregulated in the Boec group compared to the control group (P<0.05). A third experiment (experiment C) was performed to quantify and localise ADRP protein. Boec, in vivo and control groups were tested. After immunofluorescence staining followed by confocal microscopy analysis, embryonic ADRP was clearly localised around lipid droplets, indicating that ADRP is also a lipid droplet coat protein in bovine embryos. In conclusion, our results demonstrate that bovine embryos at the blastocyst stage expressed ADRP mRNA and protein, and that the embryonic culture system modified this expression.