93 Peroxisome proliferator-activated receptor delta-PPARδ agonist (L-165041) enhances bovine embryo survival and post-vitrification viability
J. A. Sánchez Viafara A I , G. Lopes de Vasconcelos A , R. Maculan B , N. Gomes Alves A , M. Brandao Dias Ferreira C , M. J. Sudano D , G. Zoccal Mingoti E , G. Barros Nunes E , R. Ribeiro de Lima A , R. Martins Drumond F , R. Nunes dos Santos F , M. A. M. Donato G , C. A. Peixoto G , J. Jasmin H and J. Camisão de Souza AA Universidade Federal de Lavras, Lavras, Minas Gerais, Brasil
B Instituto Federal do Sul de Minas, Machado, Minas Gerais, Brasil
C EPAMIG, Belo Horizonte, Minas Gerais, Brasil
D Universidade Federal do ABC, Santo André, São Paulo, Brasil
E Escola de Medicina Veterinária, Laboratório de Fisiologia da Reprodução, Universidade Estadual Paulista, Campus Araçatuba, São Paulo, Brasil
F Cenatte Embriões, Minas Gerais, Brasil
G Centro de Pesquisas Aggeu Magalhães, Fiocruz/Pernambuco, Recife, Brasil
H NUMPEX-Bio, Universidade Federal do Rio de Janeiro, Campus Duque de Caxias, Rio de Janeiro, Brasil
I Universidad de Santander, Facultad de Ciencias Agrícolas y Veterinarias, Valledupar, Colombia
Reproduction, Fertility and Development 35(2) 173-173 https://doi.org/10.1071/RDv35n2Ab93
Published: 5 December 2022
© 2023 The Author(s) (or their employer(s)). Published by CSIRO Publishing on behalf of the IETS
The effect of L-165041 (PPARδ-agonist) on decreasing apoptosis and intracellular lipid content was assessed in fresh and vitrified-warmed in vitro-produced bovine embryos. It was hypothesised that the addition of L-165041 to the culture medium enhances development and cryopreservation. At the end of the maturation period (24 h), in vitro fertilisation (IVF, Day 0) was carried out for 18–22 h. Presumed zygotes were allocated to one of two treatments: (1) control: standard culture medium, synthetic oviduct fluid medium (SOFaa), along with 2.5% FBS, 22.0 μg/mL of pyruvate, 0.025 g/mL of BSA, and 83.4 μg/mL of amikacin (n = 609), or (2) 1 µM of L-165041 (n = 608) was added to the medium on Day 1 with no medium change. Ultrastructure, cleavage, and blastocyst rates (out of total cleaved embryos) were evaluated in fresh and post-vitrification-cultured embryos. The two-step technique was used for the vitrification of blastocysts: 98 were vitrified in the control group, and 111 were vitrified in the L-165041 group. Grade 1, 2, and 3 blastocysts were washed in maintenance solution (MS) (TCM-199 HEPES + 20% FBS) and transferred to the SV1 vitrification solution (7.5% ethylene glycol + 7.5% DMSO in TCM-199 HEPES) for 3 min. Subsequently, the embryos were transferred to the SV2 vitrification solution (18% ethylene glycol + 18% DMSO + 0.4 M sucrose in TCM-199 HEPES) for 30 s. Afterward, embryos were placed on Cryotop devices (Vitri-Inga®) in groups of three to five blastocysts and immediately immersed in liquid nitrogen. A subset of fresh embryos was fixed for TUNEL assay and for Sudan-Black-B histochemical staining. Vitrified-warmed embryos were assessed using MALDI-MS technique. Cleavage and blastocyst rates (control 49.4 ± 5.2; L-165041 51.8 ± 4.3) were not influenced by L-165041. The proportion of inner cell mass cells (ICM) was higher in fresh embryos (P < 0.05), and the rate of total and ICM apoptosis was lower in L-165041 (P < 0.05). In warmed embryos, total and ICM apoptosis was lower in L-165041 (P < 0.05). The overall hatching rate, calculated for different subsets of embryos, from 6–10 embryos per treatment at each time point (12–72 h) after thawing, was higher (P < 0.001) in L-165041 (66.62 ± 2.83% vs 53.19 ± 2.9%). There was less lipid accumulation in fresh L-165041 embryos. In conclusion, the use of L-165041 reduces apoptosis, improving the viability of in vitro-derived bovine embryos.
Thanks to Professor Álan Maia Borges of the Escola de Medicina Veterinária of the Universidade Federal de Minas Gerais – UFMG, for his collaboration in supplying the experimental embryo production media.
