Simulated physiological oocyte maturation has side effects on bovine oocytes and embryos

TY - JOUR

T1 - Simulated physiological oocyte maturation has side effects on bovine oocytes and embryos

AU - Razza, Eduardo

AU - Pedersen, Hanne Skovsgaard

AU - Strøbech, Lotte

AU - Fontes, Patricia

AU - Kadarmideen, Haja

AU - Callesen, Henrik

AU - Pihl, Maria

AU - Nogueira, Marcelo

AU - Hyttel, Poul

PY - 2019/3

Y1 - 2019/3

N2 - Purpose: Oocyte maturation is a complex process involving nuclear and cytoplasmic modulations, during which oocytes acquire their ability to become fertilized and support embryonic development. The oocyte is apparently “primed” for maturation during its development in the dominant follicle. As bovine oocytes immediately resume meiosis when cultured, it was hypothesized that delaying resumption of meiosis with cyclic nucleotide modulators before in vitro maturation (IVM) would allow the oocytes to acquire improved developmental competence. Methods: We tested the Simulated Physiological Oocyte Maturation (SPOM) system that uses forskolin and 3-isobutyl-1-methylxanthine for 2 h prior to IVM against two different systems of conventional IVM (Con-IVM). We evaluated the ultrastructure of matured oocytes and blastocysts and also assessed the expression of 96 genes related to embryo quality in the blastocysts. Results: In summary, the SPOM system resulted in lower blastocyst rates than both Con-IVM systems (30 ± 9.1 vs. 35 ± 8.7; 29 ± 2.6 vs. 38 ± 2.8). Mature SPOM oocytes had significantly increased volume and number of vesicles, reduced volume and surface density of large smooth endoplasmic reticulum clusters, and lower number of mitochondria than Con-IVM oocytes. SPOM blastocysts showed only subtle differences with parallel undulations of adjacent trophectoderm plasma membranes and peripherally localized ribosomes in cells of the inner cell mass compared with Con-IVM blastocysts. SPOM blastocysts, however, displayed significant downregulation of genes related to embryonic developmental potential when compared to Con-IVM blastocysts. Conclusions: Our results show that the use of the current version of the SPOM system may have adverse effects on oocytes and blastocysts calling for optimized protocols for improving oocyte competence.

AB - Purpose: Oocyte maturation is a complex process involving nuclear and cytoplasmic modulations, during which oocytes acquire their ability to become fertilized and support embryonic development. The oocyte is apparently “primed” for maturation during its development in the dominant follicle. As bovine oocytes immediately resume meiosis when cultured, it was hypothesized that delaying resumption of meiosis with cyclic nucleotide modulators before in vitro maturation (IVM) would allow the oocytes to acquire improved developmental competence. Methods: We tested the Simulated Physiological Oocyte Maturation (SPOM) system that uses forskolin and 3-isobutyl-1-methylxanthine for 2 h prior to IVM against two different systems of conventional IVM (Con-IVM). We evaluated the ultrastructure of matured oocytes and blastocysts and also assessed the expression of 96 genes related to embryo quality in the blastocysts. Results: In summary, the SPOM system resulted in lower blastocyst rates than both Con-IVM systems (30 ± 9.1 vs. 35 ± 8.7; 29 ± 2.6 vs. 38 ± 2.8). Mature SPOM oocytes had significantly increased volume and number of vesicles, reduced volume and surface density of large smooth endoplasmic reticulum clusters, and lower number of mitochondria than Con-IVM oocytes. SPOM blastocysts showed only subtle differences with parallel undulations of adjacent trophectoderm plasma membranes and peripherally localized ribosomes in cells of the inner cell mass compared with Con-IVM blastocysts. SPOM blastocysts, however, displayed significant downregulation of genes related to embryonic developmental potential when compared to Con-IVM blastocysts. Conclusions: Our results show that the use of the current version of the SPOM system may have adverse effects on oocytes and blastocysts calling for optimized protocols for improving oocyte competence.

KW - Bovine blastocyst

KW - Gene expression

KW - In vitro maturation

KW - SPOM

KW - Ultrastructure

KW - ACTIVATION

KW - VIVO

KW - MOUSE

KW - COMPETENCE

KW - CYCLIC-AMP

KW - IN-VITRO MATURATION

KW - REDISTRIBUTION

KW - MEIOSIS

KW - CUMULUS CELLS

KW - CAMP

KW - Meiosis/genetics

KW - Oogenesis/drug effects

KW - Ovarian Follicle/drug effects

KW - Oocytes/drug effects

KW - Cattle

KW - Fertilization in Vitro/drug effects

KW - Female

KW - In Vitro Oocyte Maturation Techniques

KW - Colforsin/administration & dosage

KW - Ribosomes/drug effects

KW - 1-Methyl-3-isobutylxanthine/administration & dosage

KW - Pregnancy

KW - Animals

KW - Cumulus Cells/drug effects

KW - Embryonic Development/drug effects

KW - Blastocyst/drug effects

U2 - 10.1007/s10815-018-1365-4

DO - 10.1007/s10815-018-1365-4

M3 - Journal article

C2 - 30443692

SN - 1058-0468

VL - 36

SP - 413

EP - 424

JO - Journal of Assisted Reproduction and Genetics

JF - Journal of Assisted Reproduction and Genetics

IS - 3

ER -