TY - JOUR
T1 - Intracytoplasmic sperm injection (ICSI) of prepubertal goat oocytes using fresh and frozen-thawed semen
AU - Menéndez-Blanco, Irene
AU - Catala, Maria Gracia
AU - Roura, Montserrat
AU - Soto-Heras, Sandra
AU - Piras, Anna Rita
AU - Izquierdo, Dolors
AU - Paramio, Maria Teresa
PY - 2019/1/1
Y1 - 2019/1/1
N2 - The aim of this study was to test the effect of fresh and frozen-thawed semen on embryo development of prepubertal goat oocytes fertilized by intracytoplasmic sperm injection (ICSI). Oocytes were in vitro matured in BO-IVM medium for 26 h. Fresh and frozen-thawed semen were selected by centrifugation in BoviPure(center dot) commercial gradient. Before injection, spermatozoa were assessed for: capacitation through the detection of the tyrosine phosphorylation, viability through propidium iodide stain, and acrosomal status through fluorescein isothiocyanate-conjugated peanut agglutinin stain. Metaphase 11 oocytes were injected with a spermatozoon selected by hyaluronic-acid binding medium (SpermSlow (TM)). Results of frozen-thawed spermatozoa showed significantly higher level of capacitation than fresh spermatozoa (64.92 vs 34.67%, respectively). Moreover fresh semen group showed significantly higher (p <0.02) number of live acrosome reacted sperm (8.84%) than frozen-thawed group (6.42%). At 17 h post ICSI, zygotes (2 pronuclei) were evaluated but no differences were found using fresh and frozen-thawed semen (39.06 vs 37.50% respectively). Moreover, no differences were obtained comparing the percentage of blastocysts produced at day 9 post injection (18.46 vs 12.12%, respectively). In conclusion, either fresh or frozen-thawed semen did not have an effect on the embryo production of prepubertal goat oocytes fertilized by ICSI, despite the differences on viability, acrosomal status and sperm capacitation.
AB - The aim of this study was to test the effect of fresh and frozen-thawed semen on embryo development of prepubertal goat oocytes fertilized by intracytoplasmic sperm injection (ICSI). Oocytes were in vitro matured in BO-IVM medium for 26 h. Fresh and frozen-thawed semen were selected by centrifugation in BoviPure(center dot) commercial gradient. Before injection, spermatozoa were assessed for: capacitation through the detection of the tyrosine phosphorylation, viability through propidium iodide stain, and acrosomal status through fluorescein isothiocyanate-conjugated peanut agglutinin stain. Metaphase 11 oocytes were injected with a spermatozoon selected by hyaluronic-acid binding medium (SpermSlow (TM)). Results of frozen-thawed spermatozoa showed significantly higher level of capacitation than fresh spermatozoa (64.92 vs 34.67%, respectively). Moreover fresh semen group showed significantly higher (p <0.02) number of live acrosome reacted sperm (8.84%) than frozen-thawed group (6.42%). At 17 h post ICSI, zygotes (2 pronuclei) were evaluated but no differences were found using fresh and frozen-thawed semen (39.06 vs 37.50% respectively). Moreover, no differences were obtained comparing the percentage of blastocysts produced at day 9 post injection (18.46 vs 12.12%, respectively). In conclusion, either fresh or frozen-thawed semen did not have an effect on the embryo production of prepubertal goat oocytes fertilized by ICSI, despite the differences on viability, acrosomal status and sperm capacitation.
KW - Embryos
KW - Ruminants
KW - Sperm analysis
KW - VIVO
KW - PHOSPHORYLATION
KW - RISE
KW - EMBRYO DEVELOPMENT
KW - IN-VITRO
KW - HYALURONIC-ACID-BINDING
KW - NUCLEI
KW - BOAR SPERMATOZOA
KW - SELECTION
KW - ACROSOME REACTION
UR - http://www.mendeley.com/research/intracytoplasmic-sperm-injection-icsi-prepubertal-goat-oocytes-using-fresh-frozenthawed-semen
U2 - 10.1016/j.smallrumres.2018.11.026
DO - 10.1016/j.smallrumres.2018.11.026
M3 - Article
SN - 0921-4488
VL - 170
SP - 137
EP - 142
JO - Small Ruminant Research
JF - Small Ruminant Research
ER -