Mitochondrial organization in prepubertal goat oocytes during in vitro maturation and fertilization
Mitochondrial organization in prepubertal goat oocytes during in vitro maturation and fertilization
复制标题
青春期前山羊卵母细胞体外成熟和受精过程中的线粒体组织
DOI：
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发表时间：
2006
影响因子：
2.5
通讯作者：
M. Paramio
中科院分区：
文献类型：
--
作者：
E. Velilla;E. Rodríguez‐González;F. Vidal;D. Izquierdo;M. Paramio
本研究的目的是评估线粒体分布在体外成熟（在0，15，20，和27小时的IVM）和受精的青春期前山羊卵母细胞相比，排卵和体外受精的成年山羊卵母细胞的线粒体分布。从屠宰场回收青春期前山羊的卵母细胞，并在M199中用激素和血清成熟27小时。从促性腺激素处理的穆尔西纳山羊中收集排卵的卵母细胞。在Percoll梯度离心中选择冻融精子，并在含20%牛血清的DMH中获能1 h。将排卵和IVM‐卵母细胞在含牛血清和乳酸钙的DMH培养基中授精20 h。将卵母细胞和假定受精卵用Mitotraker绿色FM染色，并在共聚焦激光扫描显微镜下观察。透射电镜观察卵母细胞和受精卵的超微结构。青春期前山羊卵母细胞在生发泡期（GV）呈现线粒体定位于皮质和核周区。中期II的IVM-卵母细胞呈现线粒体外周极化至与中期纺锤体定位和极体内相对的区域。排卵期卵母细胞线粒体呈外周分布，线粒体聚集在MII纺锤体周围。在授精后20小时，线粒体分布在两个同步原核（2 PN rpar;在受精卵中排卵的卵母细胞中，而在青春期前的2 PN-受精卵中，线粒体呈现外周极化分布。TEM图像检测到，未成熟的青春期前的山羊卵母细胞的电密度较低，目前比在体外成熟的青春期前的山羊卵母细胞嵴少，这些特征是与肿胀的囊泡。摩尔Reprod. Dev. 73：617-626，2006 © 2006 Wiley利斯公司
The aim of this study was to evaluate mitochondrial distribution during in vitro maturation (at 0, 15, 20, and 27 hr of IVM) and fertilization of prepubertal goat oocytes compared to mitochondrial distribution of ovulated and in vitro fertilized oocytes from adult goats. Oocytes from prepubertal goats were recovered from a slaughterhouse and were matured in M199 with hormones and serum for 27 hr. Ovulated oocytes were collected from gonadotrophin‐treated Murciana goats. Frozen‐thawed spermatozoa were selected by centrifugation in Percoll gradient and were capacitated in DMH with 20% steer serum for 1 hr. Ovulated and IVM‐oocytes were inseminated in DMH medium with steer serum and calcium lactate for 20 hr. Oocytes and presumptive zygotes were stained with Mitotraker Green FM and observed under a confocal laser scanning microscope. Ultrastructural morphology of oocytes and presumptive zygotes were analyzed by transmission electron microscopy (TEM). Prepubertal goat oocytes at germinal vesicle stage (GV) presented mitochondria localized in the cortical and perinuclear region. IVM‐oocytes at metaphase II presented mitochondria peripheral polarized to the region opposite were the metaphase spindle is positioned and within the polar body. Ovulated oocytes presented peripheral mitochondria distribution and mitochondrial aggregation around the MII spindle. At 20 hr post‐insemination, mitochondria were distributed around the two synchronous pronuclei (2PN rpar; in zygotes ovulated oocytes whereas in prepubertal 2PN‐zygotes mitochondria presented a peripheral polarized distribution. Images by TEM detected that immature prepubertal goat oocytes that are less electrodense and present fewer cristae than in vitro matured prepubertal goat oocytes; these are characterized by being associated to swollen vesicles. Mol. Reprod. Dev. 73: 617–626, 2006 © 2006 Wiley‐Liss, Inc.
影响因子：
13.3
作者：
Bavister, BD
通讯作者：
Bavister, BD