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Standard patch-clamp technique was used to study the allosteric modulation of ATP sensitive by intracellular pH.

方法用人胚肾细胞表达Kit6.2野生型、R371H和P266T位点突变后的通道,用膜片钳技术研究突变后胞内pH对ATP敏感性变构调节的变化。

Ovary globose or ovoid; ovule 1, pendulous; style short or long; stigma capitate. Utricles globose or ovoid, indehiscent, falling off with perianth.

子房球状或卵球形;胚珠1,下垂;花柱短或长;柱头头状胞果球状或卵球形,不裂,与花被一同脱落。

Results: Embryoid bodies grew well in Collagen Gel, the differentiated cells grew into the gel gradually; electronmicroscope showed that some induced cells were rich in Golgi's bodies and endoplasmic reticulums, a few secretory granules were also visible, which was similar to human adult thyrocyte.

结果 拟胚体在鼠尾胶原上生长状态良好,分化细胞贴壁后逐渐陷入鼠尾胶原;电镜下诱导细胞高尔基体、内质网丰富,部分诱导细胞胞质内见不典型分泌颗粒,与人成体甲状腺细胞超微结构类似。

In conclusions,(1) The pronuclear formation was asynchronous after ICSI in buffalo, the female pronuclear formed 3 hours earlier than male pronuclei;(2) After ICSI and activation, Culture of oocytes in 1.9mmol/L 6-DMAP for 3 hours can improve their subsequent embryonic development;(3) Treatment of sperm with 5mmol/L DTT can promote sperm decondensation;(4) Dead sperms can be used for ICSI in buffaloes;(5) Treatment of sperm with GSH can improve the efficiency of ICSI in buffaloes.

以上结果表明:(1)水牛精子胞质内显微受精的雌、雄原核发育不同步,雌原核比雄原核早3h形成;(2)水牛卵母细胞ICSI和激活后用1.9mmol/L 6-DMAP培养处理3h能提高其胚胎发育率;(3)DTT预处理水牛精子能提高其ICSI后的精子解聚率;(4)死精子能用于水牛卵母细胞的ICSI;(5)GSH预处理水牛精子有助于提高其ICSI后的囊胚发育率。

In the present study, we collected cumulus cells oocyte complex from ovaries of two different strain mice. The cumulusenclosed oocytes were cultured for 6 h in MEM supplemented with growth factor and FSH. The meiotic maturation of these oocytes has progressed to pro-metaphse Ⅰ stage and the condensed chromosomes are visible under DIC microscope, metaphase Ⅰ spindle even can be detected under Polscope. The metaphase Ⅰ spindles of oocytes were exchanged under such microscopes. After electric stimuli, 91. 6% and 91. 6% karyoplasts-cytoplasm pairs were fused respectively. The resulting oocytes were cultured further in MEM and over 80% of oocytes released the first polar body. 79% and 77% of oocytes formed two pronuclei after in vitro fertilization and the embryos were cultured in KSOM supplemented with amino acids. Over 60% of embryos developed to blastocyst stage.

在本研究中我们在取得两种不同品系小鼠的卵丘卵母细胞复合体后,先将卵丘卵母细胞复合体置于含有多种生长因子和激素的MEM培养液中培养6小时,此时卵母细胞已进入第一次减数分裂的前中期,并且在DIC倒置显微镜下可以看到浓缩的染色体,用Polscope可以发现明显的纺锤体,借助这种显微镜通过显微操作将两种不同品系小鼠来源的卵母细胞的MI纺锤体进行互换,经过三次直流电脉冲作用后,分别有91.6%的胞质—MI核质体对融合,经过进一步的培养后,超过80%的重组卵母细胞排出第一极体,体外受精后分别有79%和77%的重组卵形成双原核,受精后的胚胎在KSOM胚胎培养液中体外培养4天后,超过60%的胚胎发育至囊胚。

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