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Parthenogenetic Activation of Pig Oocytes: RESULTS(4)

As shown in Table 3, when oocytes treated with A23187 were inseminated after culture for 3.5 h (experiment 2), the penetration rate (12.0 ± 9.6%) was the same as that after culture for 10 min (12.2 ± 12.7%), and these rates were significantly lower than those in controls (82.4 ± 8.6-85.2 ± 9.7%). The proportion of oocytes with male pronuclei was lower in A23187-treated oocytes (63.3 ± 29.6%) that had been cultured for 3.5 h before insemination than in control oocytes (88.1 ± 6.3%). Polyspermic penetrations in oocytes were significantly lower in A23187-treated oocytes than in control oocytes. ventolin inhalers

Penetrability of Plasma Membrane of Pig Oocytes Activated by A23187

As shown in Table 4, when A23187-treated oocytes were inseminated after ZP were removed, the same (p > 0.05) rate of sperm penetration was observed as that in untreated ZP-free oocytes. However, as in experiment 1, ZP-intact oocytes showed a significantly lower penetration rate (1.3 ± 2.2%) in A23187-treated oocytes than in control oocytes (88.3 ± 12.0%). High proportions (85.3 ± 9.3-100%) of oocytes also formed a male pronucleus regardless of A23187 treatment. Polyspermic penetration was significantly higher in ZP-free than in ZP-intact oocytes. More spermatozoa were also observed in ZP-free (2.6 ± 1.5-2.7 ± 1.4/oocyte) than in ZP-intact oocytes (1.0 -1.5 ± 0.7/ oocyte).

TABLE 3. Penetration of zona-intact pig oocytes treated with A23187 before insemination.*

Culture time beforeinsemination A23187 No. of oocytes examined No. of oocytes penetrated(%Y No. of oocytes with FPN+(%)a No. of oocytes with MPN*(%)b No. of polyspermic oocytes(%)b
10 min 247 210 (85.2 ± 9.7)’ 210 (85.2 ± 9.7)’ 192 (91.7 ± 8.5)’ 129 (65.6 ± 14.6)’
+ 219 28 (12.2 ± 12.7)d 157 (72.0 ± 20.6)’d 23 (73.6 ± 25.6)’d 3 (4.6 ± 10.2)d
3.5 h 257 212 (82.4 ± 8.6)’ 212 (82.4 ± 8.6)’ 188 (88.1 ± 6.3)’ 120 (59.3 ± 15.2)c
+ 242 32 (12.0 ± 9.6)d 143 (60.8 ± 1 7.2)d 19 (63.3 ± 29.6)d 7 (20.7 ± 20.1 )d

*    Oocytes were treated with 100 |xM A23187 for 5 min and then cultured for 10 min or 3.5 h before IVF. Experiments were repeated nine times. The data were expressed as mean ± SD.
+ FPN: female pronucleus.
*    MPN: male pronucleus.

TABLE 4.

Treatment of oocytes by A23187 Presence (+) or absence of ZP at IVF No. of oocytes examined No. of oocytes penetrated(%)a No. of oocytes with FPN*(%Y No. of oocytes with MPNS(%)b No. of polyspermic oocytes(%)b Average no. of spermatozoa in penetrated oocytes
+ * + 65 1 (1.3 ±2.2)’ 46(72.3 ± 12.5) 1 0′ 1.0 ± 0.0′
+ * _t 52 48 (92.7 ± 1,5)d 46 (88.7 ± 4.6) 43 (90.0 ± 5.3) 34 (69.3 ± 13.4)d 2.7 ± 1.4d
+ 77 67 (88.3 ± 12.0)d 67 (87.3 ± 7.1) 66 (98.7 ± 2.3) 22 (32.6 ± 3.1)’ 1.5 ± 0.7е
50 44(89.7 ± 11.7)d 38 (76.0 ± 9.5) 38 (85.3 ± 9.3) 30(67.3 ± 8.1 )d 2.6 ± 1.5d

*    Oocytes (cultured for 44 h) were treated with 100 jjlM A23187 for 5 min before IVF. Experiments were repeated four times. The data were expressed as mean ± SD.
f Zona pellucida (ZP) was removed by exposure to 0.1% pronase.
*    FPN: female pronucleus. s MPN: male pronucleus.