Effect of Glycosaminoglycans on <italic>In vitro</italic> Fertilizing Ability and <italic>In vitro</italic> Developmental Potential of Bovine Embryos

Abstract

The glycosaminoglycans (GAGs) present in the female reproductive tract promote sperm capacitation. When bovine sperm were exposed to 10 μg/ml of one of four GAGs (Chondroitin sulfate, CS; Dermatan sulfate, DS; Hyaluronic acid, HA; Heparin, HP) for 5 h, the total motility (TM), straight-line velocity (VSL), and curvilinear velocity (VCL) were higher in the HP- or HA-treated sperm, relative to control and CS- or DS-treated sperm. HP and HA treatments increased the levels of capacitated and acrosome-reacted sperm over time, compared to other treatment groups (p<0.05). In addition, sperm exposed to HP or HA for 1 h before IVF exhibited significantly improved fertilizing ability, as assessed by 2 pronucleus (PN) formation and cleavage rates at d 2. Exposure to these GAGs also enhanced in vitro embryo development rates and embryo quality, and increased the ICM and total blastocyst cell numbers at d 8 after IVF (p<0.05). A real-time PCR analysis showed that the expression levels of pluripotency (Oct 4), cell growth (Glut 5), and anti-apoptosis (Bax inhibitor) genes were significantly higher in embryos derived from HA- or HP-treated sperm than in control or other treatment groups, while pro-apoptotic gene expression (caspase-3) was significantly lower in all GAG treatment groups (p<0.05). These results demonstrated that exposure of bovine sperm to HP or HA positively correlates with in vitro fertilizing ability, in vitro embryo developmental potential, and embryonic gene expression.

Post-Death Cloning of Endangered Jeju Black Cattle (Korean Native Cattle): Fertility and Serum Chemistry in a Cloned Bull and Cow and Their Offspring

“…Two microliters each of heparin (2 µg/ml) and PHE (18.2 M penicillamine, 9.1 M hypotaurine and 1.8 M epinephrine) were also added into the 44-µl IVF drop. Sperm was prepared using the two-step swim-up protocol (patent no: KR101064415) [16]. To remove the egg yolk, thawed semen was slowly added to 3 ml of 20% Triladyl ® solution (Triladyl ® :distilled water = 1:4) and centrifuged at 200 g (2000 rpm) for 1 min.…”

Evaluation of Semen Quality of Jeju Black Cattle (JBC) to Select Bulls Optimal for Breeding and Establish Freezing Conditions Suitable for JBC Sperm

“…Sperm propulsion is caused by the active motion of their flagella. Sperm quality has been assessed by measuring sperm motility, including measurements of VCL, VSL, and VAP [ 14 , 15 , 16 , 17 , 18 ]. The mean VCL, VSL, and VAP of sperm from Karan Fries bulls, a composite Indian breed, were found to be 188.63 ± 23.4 μm/s, 81.39 ± 16.2 μm/s, and 102.58 ± 20.5 μm/s, respectively [ 18 ].…”