An intravital microscopy method permitting continuous long-term observations of ovulation in vivo in the rabbit
An intravital microscopy method permitting continuous long-term observations of ovulation in vivo in the rabbit
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DOI：
10.1093/humrep/dei394
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发表时间：
2006-03-01
影响因子：
6.1
通讯作者：
Brännström, M
中科院分区：
文献类型：
--
作者：
Dahm-Kähler, P;Löfman, C;Brännström, M
背景：开发了一种家兔卵巢活体显微镜检查方法，以观察体内排卵期间的实时变化。目的是将这些事件与排卵特定阶段的生化事件相关联。方法：用马绒毛膜促性腺激素（CG）30-100 IU，2d后再用HCG 100 IU诱导排卵。在麻醉期间，通过手术将右侧卵巢取出并浸没在器官室中，显微镜透镜靠近卵巢放置。进行连续视频记录。结果：初始马CG引发实验显示最高的排卵率，没有过早的黄体化，30 IU马CG后。该预充方案随后显示HCG后11.5-14小时卵泡破裂。外置卵巢和对侧非外置卵巢的排卵数相似。排卵的典型特征依次为：卵泡顶端微循环关闭，卵泡壁出现瘀点，破裂处形成锥形结构，破裂出血明显，颗粒细胞和卵母细胞挤出速度稳定。结论：该方法捕获了排卵过程中的一系列结构变化。它可以与血液和卵泡液采样结合进行生化分析，并可用于研究排卵期间卵泡结构特定变化相关的生化反应。
BACKGROUND: A method for intravital microscopy of the rabbit ovary was developed to enable observations of real-time changes during ovulation in vivo. The aim was to correlate these events to biochemical events at specific stages of ovulation. METHODS: Virgin, female rabbits were primed with equine chorionic gonadotrophin (CG) (30-100 IU) then HCG (100 IU) 2 days later to induce ovulation. During anaesthesia, the right ovary was surgically exteriorized and submerged in an organ chamber with a microscopy lens positioned close to the ovary. Continuous video recordings were performed. RESULTS: Initial equine CG priming experiments revealed the highest ovulation rate, without premature luteinization, after 30 IU equine CG. This priming protocol subsequently demonstrated follicular ruptures 11.5-14 h after HCG. Numbers of ovulations from the exteriorized and contralateral non-exteriorized ovary were similar. The sequence of typical features of ovulation was: shutdown of microcirculation in the follicular apex, formation of petechiae in the follicular wall and a cone-shaped structure over the future rupture site, marked bleeding in connection with follicular rupture and a fairly steady extrusion velocity of granulosa cells and the oocyte. CONCLUSION: This method captured a sequence of structural changes during ovulation. It could be combined with blood and follicular fluid sampling for biochemical analysis and could be used in studies on biochemical reactions in relation to specific changes in the follicular structure during ovulation.