Repositorio
Institucional

Salamone, Daniel Felipe

Gabriela Canel, Natalia#Suva, Mariana#Jimena Bevacqua, Romina#Elena Arias, Maria#Felmer, Ricardo

Improved embryo development using high cysteamine concentration during IVM and sperm co-culture with COCs previous to ICSI in bovine

ELSEVIER SCIENCE INC

THERIOGENOLOGY

en

In contrast to other species, intracytoplasmic sperm injection (ICSI) in bovine remains inefficient, resulting in low embryo developmental rates. It is unclear whether such inefficiency is due to the poor response of bovine ooplasms to the injection stimulus, or to the inability of bull sperm to induce oocyte activation. In order to facilitate these events, two strategies were assessed: the use of high concentration of cysteamine [Cys] during IVM, and the selection of sperm attached to cumulus cells after incubation with COCs for ICSI. First, COCs were IVM with increasing [Cys] and subjected to IVF. Zygotes from all groups were cultured under different O-2 tensions and development to blastocyst was evaluated. In a second experiment, sperm were co-cultured for 3 h with COCs and acrosome reaction was studied. Afterwards, the best IVM and IVC conditions determined on Experiment I were used for ICSI assay. COCs were matured for 21 h with I (Cys 1) or 0.1 mM Cys (Cys 0.1 groups, standard condition). In addition, COCs were incubated for >= 3 h with 16 x 10(6) spermiml and only sperm attached to cumulus cells were selected for ICSI (ICSI + Co-cult groups). After chemical activation, embryos were cultured in SOF medium under low O-2 tension. Cleavage and blastocyst rates were evaluated at days 2 and 7 of IVC, respectively. Finally, the relative expression of eight genes indicators of embryo quality was compared between ICSI and IVF control blastocysts by qPCR. Cleavage rates were higher for Cys 0.1 ICSI + Co-cult and Cys 1 ICSI + Co-cult groups (n = 117, 92% and n = 116, 79%, respectively) compared to their controls (n = 132, 60% for Cys 0.1 ICSI and n = 108, 52% for Cys 1 ICSI) (p <= 0.05). Interestingly, the combined treatment (Cys 1 ICSI + Co-cult) showed higher blastocyst rates than all other ICSI groups (23 vs. 11, 18 and 14% for Cys 0.1 ICSI + Co-cult, Cys 1 ICSI, and Cys 0.1 ICSI, respectively) (p <= 0.05). Moreover, incubation with COCs increased the rates of live acrosome reacted sperm (p <= 0.05). The relative abundance of mRNA5 coding for INFr, CAT, DNMTI, OCT4, and HDAC3 did not differ between treatments (p <= 0.05). SOD2, HADC1 and HADC2 expression was higher for Cys 0.1 ICSI than for IVF embryos (p <= 0.05). Group Cys1 ICSI did not differ from IVF for those three genes, neither did Cys 1 ICSI + Co-cult, except for HDACI (p <= 0.05). In conclusion, the use of 1 mM Cys during IVM and of sperm incubated with mature COCs might be a good strategy to improve ICSI outcomes in cattle. (C) 2018 Published by Elsevier Inc.

Artículo original

This work was supported by the grant from Agencia Nacional de Promocion Cientifica y Tecnologica PICT-2010-2716 from the Ministry of Science and Technology of Argentina.

Este trabajo fue apoyado por la subvención de la Agencia Nacional de Promoción Científica y Tecnológica PICT-2010-2716 del Ministerio de Ciencia y Tecnología de Argentina.

10.1016/j.theriogenology.2018.05.017

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Sperm injection# Cumulus cells# Pretreatment# In vitro maturation# Oxygen tension# Gene expression

http://dx.doi.org/10.1016/j.theriogenology.2018.05.017

Reproductive Biology# Veterinary Sciences

Inyección de esperma# Células de cúmulo# Pretratamiento# Maduración in vitro# Tensión de oxígeno# expresión genética

Ciencia Animal y lechería

Biología Reproductiva

Biotecnología agrícola

WOS:000436055900004

Improved embryo development using high cysteamine concentration during IVM and sperm co-culture with COCs previous to ICSI in bovine

hdl.handle.net/11336/91372

Artículo original

version publicada

ELSEVIER SCIENCE INC

THERIOGENOLOGY

Biología Reproductiva# Ciencias Veterinarias

0093-691X

en

ANPCyT PICT-2010-2716

ANICyT Argentina PICT-2010-2716

This work was supported by the grant from Agencia Nacional de Promocion Cientifica y Tecnologica PICT-2010-2716 from the Ministry of Science and Technology of Argentina.

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