选择对异氟烷敏感性不同的小鼠品系进行选择性繁殖。

Selective breeding of mice strains with different sensitivity to isoflurane.

机构信息

Department of Critical Care Medicine, Beijing Tiantan Hospital, Capital Medical University, Beijing 100050, China.

出版信息

Chin Med J (Engl). 2010 May 20;123(10):1315-9.

PMID:20529588

Abstract

BACKGROUND

The mechanisms of action of volatile anesthetics are still unknown. Recently, the use of genetics as a means to investigate anesthetic action has increased in scale. However, only limited forward genetic approach studies were performed in mammals, especially with volatile anesthetics as the selection agent. In the present study, a selective breeding process was designed to produce strains of mice with different sensitivity to isoflurane.

METHODS

One hundred and sixty male and female virgin outbred ICR/CD-1 mice at 65 - 70 days of age were selected as original generation, and the median effective dose (ED(50)) of inhaled isoflurane were measured by probit analysis with the loss of righting reflex as the endpoint of anesthesia. The most sensitive males and females were selected and mated one another randomly, as with the most resistant males and females. Thus two branches of mice (sensitive and resistant to isoflurane) were created and allowed to produce the next generation. At 65 - 70 days of age, screening experiment was performed in offspring, by selecting the most sensitive mice in sensitive branch and the most resistant mice in resistant branch. Selected males and females within each branch were mated one another randomly to produce the following generation. The same procedure was performed in the offspring. The process of screening and breeding was repeated for 8 generations, and then strains were conserved by mating the offspring one another randomly within each branch for 3 generations. Each pair of mice was allowed to produce the second litters as a backup, and isoflurane ED(50) was measured in mice from the second litters.

RESULTS

Isoflurane righting reflex ED(50)s (95% confidence limit (CL)) in original mice were 0.65% (0.58% - 0.72%) in females and 0.63% (0.56% - 0.69%) in males. After the 4th generation, isoflurane ED(50)s in resistant branch were significantly higher than those in sensitive branch (P < 0.05), for both in females and males. In the 11th generation, isoflurane ED(50) in the two branches differed by 32% in females and 36% in males.

CONCLUSIONS

After 8 generations of selective breeding and 3 generations of strain conservation, two strains of mice with high and low sensitivity to isoflurane were developed. The separation of inhaled anesthetic requirement in parents could be transferred to the offspring in mice.

摘要

背景

挥发性麻醉剂的作用机制仍不清楚。最近，利用遗传学作为研究麻醉作用的手段在规模上有所增加。然而，只有有限的正向遗传方法研究在哺乳动物中进行，特别是以挥发性麻醉剂作为选择剂。在本研究中，设计了一个选择性繁殖过程，以产生对异氟醚敏感性不同的小鼠品系。

方法

选择 160 只 65-70 日龄的雄性和雌性处女远交 ICR/CD-1 小鼠作为原始种群，并通过概率分析测量吸入异氟醚的半数有效剂量（ED(50)），以失去翻正反射作为麻醉终点。选择最敏感的雄性和雌性进行随机交配，最敏感的雄性和雌性进行随机交配。这样就创建了两个分支的小鼠（对异氟醚敏感和不敏感），并允许它们产生下一代。在 65-70 日龄时，通过在敏感分支中选择最敏感的小鼠，在抗性分支中选择最抗性的小鼠，对后代进行筛选实验。在每个分支中，选择的雄性和雌性随机交配产生下一代。在后代中重复进行相同的筛选和繁殖过程。筛选和繁殖过程重复进行 8 代，然后通过在每个分支内随机交配，使品系保存 3 代。每对小鼠允许产生第二胎作为备用，在第二胎中测量异氟醚 ED(50)。

结果

原始小鼠的异氟醚翻正反射 ED(50)s（95%置信区间（CL））为雌性 0.65%（0.58%-0.72%），雄性 0.63%（0.56%-0.69%）。在第 4 代后，抗性分支的异氟醚 ED(50)s 显著高于敏感分支（P<0.05），雌性和雄性均如此。在第 11 代，两个分支的异氟醚 ED(50)在雌性中相差 32%，在雄性中相差 36%。