RT Journal Article
T1 Acquisition of fertilization competence in guinea pig spermatozoa under different capacitation protocols
A1 Cañón Beltrán, Karina Esperanza
A1 Cajas Suárez, Yulia Nathaly
A1 González Martínez, María Encina
A1 Fernández-González, Raul
A1 Fierro, Natacha
A1 Lorenzo González, Pedro Luis
A1 Arias Álvarez, María
A1 García García, Rosa María
A1 Gutiérrez Adán, Alfonso
A1 Rizos, Dimitri
AB Guinea pig in vitro fertilization (IVF) are poorly developed due to the limited accessibility to oocytes and the lack of an efficient method of sperm capacitation. Thus, we aimed to evaluate different capacitation protocols that we validated through sperm analysis and using heterologous (He) IVF with zona-intact bovine oocytes. Spermatozoa of guinea pigs were collected and processed separately by 4 different protocols: A) Spermatozoa were obtained by flushing the lumen of one cauda epididymis and incubated in a minimal culture medium (MCM); B) One epididymis was placed in a prewarmed of M2 medium and gently minced with fine scissors. Spermatozoa were incubated in a modified human tubal fluid medium (HTF). In both protocols, the spermatozoa were capacitated at 37 °C under an atmosphere of 5% CO2 for 2 h. In the protocols C and D, the spermatozoa were collected by flushing the lumen of the cauda epididymis and selected by commercial density gradient Bovipure® (Nidacon Laboratories AB, Göthenborg, Sweden), according to the manufacturer's instructions. Then for Protocol C) spermatozoa were incubated in MCM medium supplemented with 10 mg/mL heparin (MCM-Hep); while for Protocol D) spermatozoa were incubated in FERT medium supplemented 10 mg/mL heparin (FERT-Hep). Incubation of C and D protocols were performed at 38.5 °C under an atmosphere of 5% CO2 for 2 h. Capacitation protocols C and D showed a higher percentage of viability, total and hyperactive-like motility, and acrosome reaction compared to protocols A and B. For this reason, protocols C and D were used for further He-IVF analysis. Guinea pig sperm and matured zona-intact bovine oocytes were co-incubated at 5% CO2 and 38.5 °C. Sperm-oocyte interaction was assessed at 2.5 h post-insemination (hpi) and pronuclear formation (PrF) were evaluated at 18, 20, 22, 24 and 26 hpi, while the cleavage rate was evaluated at 48 hpi. In protocol D, PrF was significantly higher than in protocol C (P ≤ 0.05) at every time point evaluated. Also, the cleavage rate at 48 hpi was higher (P ≤ 0.05) in He-IVF protocol D (69.8 ± 1.7%) compared to He-IVF protocol C (49.1 ± 1.1%). In conclusion, we determined the most adequate sperm capacitation conditions for guinea pig that allow zona-intact bovine oocyte penetration and lead to hybrid embryo formation, suggesting that these conditions could be optimal to develop IVF in guinea pigs.
PB Elsevier
SN 0093-691X
YR 2023
FD 2023-03-01
LK https://hdl.handle.net/20.500.14352/105623
UL https://hdl.handle.net/20.500.14352/105623
LA eng
NO Karina Cañón-Beltrán, Yulia N. Cajas, Encina González, Raúl Fernández-González, Natacha Fierro, Pedro L. Lorenzo, María Arias-Álvarez, Rosa M. García-García, Alfonso Gutiérrez-Adán, Dimitrios Rizos, Acquisition of fertilization competence in guinea pig spermatozoa under different capacitation protocols, Theriogenology, Volume 198, 2023, Pp 231-240, https://doi.org/10.1016/j.theriogenology.2022.12.042.
NO Ministerio de Ciencia e Innovación (España)
DS Docta Complutense
RD 8 ago 2024