Appendix 1—table 6. Rates of sperm penetration and binding to the bovine ZP after homologous (bovine sperm) and heterologous (human or murine sperm) EZPT using bovine ZPs co-incubated without bovine oviductal fluid (HoA, HeA) or with bovine oviductal fluid (HoB, HeB).
| Group N | Sperm | Oviductal fluid | 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 90 |
Bovine | - | 4.16±0.27 | 90 (100±0)a |
853 (9.13±1.23)a |
30 ± 1.41a | 90 (100±0)a |
830 (9.27±0.58)a |
23 ± 1.12a |
| HeAh 406 |
Human | - | 9.14±3.98 | 283 (69.7±12.1)b |
1588 (5.61±2.83)b |
23±4.2b | 96 (23.65±8.59)b |
188 (1.49±0.24)b |
6±1.27b |
| HeAm 83 |
Murine | - | 8.76±1.07 | 80 (96.25±5.3)a |
300 (3.73±0.31)C |
21±2.12b | 83 (100±0)a |
490 (5.92±0.81)c |
20 ± 3.54a |
| HoB 56 |
Bovine | + | 4.16±0.27 | 56 (100±0)a |
585 (10.52±0.56)a |
24±2.82b | 56 (100±0)a |
574 (10.02±1.81)a |
23 ± 2.12a |
| HeBh 131 |
Human | + | 9.14±3.98 | 0 (0±0)c |
0 (0±0)c |
0 ± 0c | 10 (7.42±2.53)c |
10 (1±0)d |
1 ± 0c |
| HeBm 48 |
Murine | + | 9.71±0.61 | 4 (0.08±0.005)c |
4 (1±0)c |
1 ± 0c | 20 (41.74±2.46)b |
45 (0.94±0.08)d |
5±0.71b |
HoA = homologous EZPT with bovine sperm. HeAh = heterologous EZPT with human sperm. HeAm = heterologous EZPT with murine sperm. HoB = homologous EZPT with bovine sperm (ZPs were co-incubated with bovine oviductal fluid for 30 min). HeBh = heterologous EZPT with human sperm (ZPs were co-incubated with bovine oviductal fluid for 30 min). HeBm = heterologous EZPT with murine sperm (ZPs were co-incubated with bovine oviductal fluid for 30 min). 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 ZPs 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.