Kure-bayashi S, Miyake M, Okada K, Kato S
The Graduate School of Science and Technology, Kobe University, Hyogo, Japan.
Theriogenology. 2000 Mar 15;53(5):1105-19. doi: 10.1016/S0093-691X(00)00256-9.
In the present study, we derived parthenogenetic porcine fetuses from in vitro-matured oocytes following a simple activation process in order to evaluate their developmental limitations in-vivo. Follicular oocytes collected from gilts at local slaughterhouses were matured for 48 h. They were subjected to a single square pulse of direct current for 100 microsec at 1,500 V/cm and then treated with 5 microg/mL cytochalasin B for 4 h to obtain activated diploid oocytes. The diploids were cultured in modified Whitten's medium until transfer. Diploids which had cleaved to the 2- and 3- to 4-cell stages were transferred to oviducts of recipients. Live and/or dead parthenogenetic fetuses were recovered in 6 of 8 trials at 17, 18, 19, 24 and 29 d post activation. The total proportion of fetuses to transferred diploids was 31.3% (62/198). When fetuses were recovered at 19 d post activation, the proportion of development into fetuses was 71% (15/21). Our results, however, suggest that periods of gestation longer than 19 d resulted in a decrease of these proportions to 45% (18/40) at 24 d and to 18% (7/40) at 29 d. The hearts were beating in nearly all of the fetuses recovered at 19, 24 and 29 d post activation. Thus, parthenogenetic porcine diploids developed to at least the stage of limb-bud formation beyond the early heart-beating stage. Abnormalities were also externally visible on some fetuses. Formation of cyst-like structures in the heart and liver, and insufficient development of the head region and acephali were observed in some cases.
在本研究中，我们通过简单的激活过程从体外成熟的卵母细胞中获得孤雌生殖的猪胎儿，以评估它们在体内的发育局限性。从当地屠宰场的后备母猪收集的卵泡卵母细胞成熟48小时。将它们在1500V/cm下施加100微秒的单方形直流电脉冲，然后用5μg/mL细胞松弛素B处理4小时以获得激活的二倍体卵母细胞。二倍体细胞在改良的惠滕氏培养基中培养直至移植。已分裂至2细胞和3至4细胞阶段的二倍体细胞被移植到受体的输卵管中。在激活后17、18、19、24和29天的8次试验中的6次中回收了活的和/或死的孤雌生殖胎儿。胎儿与移植的二倍体细胞的总比例为31.3%（62/198）。当在激活后19天回收胎儿时，发育成胎儿的比例为71%（15/21）。然而，我们的结果表明，妊娠期超过19天会导致这些比例在24天时降至45%（18/40），在29天时降至18%（7/40）。在激活后19、24和29天回收的几乎所有胎儿中，心脏都在跳动。因此，孤雌生殖的猪二倍体至少发育到了早期心跳阶段之后的肢芽形成阶段。在一些胎儿上也能从外部观察到异常。在某些情况下，观察到心脏和肝脏中形成囊状结构，以及头部区域发育不全和无头畸形。
