Appendix 1—table 14. Rates of sperm penetration and binding to bovine ZP after homologous (bovine sperm) and heterologous (murine sperm) EZPT using bovine ZPs co-incubated with bovine OVGP1 previously treated with Neuraminidase (NMase) (HoA, HeA) and with non-treated bOVGP1 (HoB, HeB).
Control groups without ZPs treatment was added for both, homologous (HoC) and heterologous (HeC).
| Group N | Bovine OVGP1 | NMase treatment to bOVGP1 | TMC | Penetrated ZPs (%) | Sperm penetration inside ZP (mean) | Max. no. of sperm inside a single ZP | ZP with bound sperm (%) | Sperm bound to ZPs (mean) | Max. no. of sperm bound to a single ZP |
|---|---|---|---|---|---|---|---|---|---|
| HoA 15 |
+ | + | 5.25±0.60 | 76 (84.49±9.91)a |
473 (5.11±1.12)a |
22±1.53 | 87 (100±0) |
720 (8.02±1.06)a |
29±3.05 |
| HoB 87 |
+ | - | 5.25±0.60 | 15 (100±0)b |
254 (21.07±7.61)b |
29±3.05 | 15 (100±0) |
355 (23.67±7.11)b |
33±5.51 |
| HeA 82 |
+ | + | 5.34±0.05 | 45 (52.78±12.73)c |
123 (1.46±0.27)c |
6.00±1.00 | 65 (79.58±7.88) |
286 (3.51±0.31)c |
11±1.52 |
| HeB 15 |
+ | - | 5.34±0.05 | 0 (0±0)d |
0 (0±0)d |
0±0 | 4 (26.67±11.54) |
5 (0.33±0.23)d |
2±0.57 |
| HeC 20 |
- | - | 5.34±0.05 | 18 (88.10±6.73)a |
113 (5.70±0.19)a |
13±2.12 | 20 (100±0) |
185 (9.98±2.61)a |
19±3.54 |
HoA = homologous EZPT with bovine sperm (ZPs were co-incubated with bovine OVGP1 for 30 min. Bovine OVGP1 was previously treated by incubation with NMase at 10U/mL in acetate buffer, PH 4.5 at 38ºC for 18 hr). HoB = homologous EZPT with bovine sperm (ZPs were co-incubated with bovine OVGP1 for 30 min. Bovine OVGP1 was previously incubated in acetate buffer, PH 4.5 at 38ºC for 18 hr). HeA = heterologous EZPT with murine sperm (ZPs were co-incubated with bovine OVGP1 for 30 min. Bovine OVGP1 was previously treated with NMase at 10U/mL in acetate buffer, PH 4.5 at 38ºC for 18 hr). HeB = Heterologous EZPT with murine sperm (ZPs were co-incubated with bovine OVGP1 for 30 min. Bovine OVGP1 was previously treated in acetate buffer, PH 4.5 at 38ºC for 18 hr). HeC = heterologous EZPT with murine sperm (ZPs were co-incubated with acetate buffer, PH 4.5 at 38ºC for 18 hr). TMC = total motile sperm count after capacitation calculated by multiplying the volume by concentration (million sperm/mL) and by the motility (%) after capacitation. Penetrated ZPs refers to the mean number of ZPs penetrated by at least one sperm. ZPs with bound sperm was calculated as the mean number of ZP with, at least, one bound sperm. Sperm penetration was taken as the total number of sperm observed inside the zonas. Bound sperm was expressed as the mean number of sperm that remained bound to the ZP. ‘N’ refers to the total number of ZPs fertilized per treatment. Analysis was conducted by using a one-way ANOVA, followed by Tukey´s post hoc test. Different superscripts (a, b, c, d) in the same column indicate significant differences (p <0.05; n=3). Values are expressed as the mean ± SD.