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Vasoligation in Mongolian Gerbils   

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摘要:雄沙鼠输精管结扎是建立假孕雌性沙鼠和胚胎移植等工作的基础。我们综合考虑了年龄等因素,通过实验验证制定本技术规程。结果表明,年龄8周以上是结扎长爪沙鼠输精管理想的时间段,在结扎后至1年内可获得较好的交配和假孕效果,为后续进行胚胎移植等工作奠定了良好基础。

关键词: 长爪沙鼠, 输精管结扎术, 年龄

材料与试剂

  1. 35 mm无菌塑料培养皿(Corning,catalog number: 430165)
  2. 医用缝合线(上海浦东金环医疗用品股份有限公司)
  3. 麻醉剂Avertin(sigma)

仪器设备

  1. CO2培养箱(Thermo, Herocell 180)
  2. 手术器械(手术剪,解剖剪,眼科镊,显微镊,显微剪)
  3. 体视显微镜(Motic SN-168)

实验步骤

  1. 麻醉将长爪沙鼠称重,腹腔注射麻醉剂Avertin (sigma, 0.12-0.15 mL/10 g), 待雄鼠进入深度麻醉状态之后再进行以下操作。
  2. 沙鼠的保定和被毛 将长爪沙鼠腹面向上放在手术台上,剪去腹部被毛;用75%酒精擦拭长爪沙鼠腹部,消毒。
  3. 在雄性长爪沙鼠下肢上端水平线正中用剪刀横向剪开1.5 cm,用组织镊夹住肌肉层, 配合眼科镊撕开相同大小的开口。
  4. 用眼科镊向下方夹住一侧的脂肪垫将睾丸、附睾和输精管拉出切口。
  5. 找到输精管,钝性分离输精管约1 cm。
  6. 左手拿眼科镊将分离后的输精管轻轻夹起,右手拿弯镊,在酒精灯上将弯镊前端烧红之后,将夹住输精管的眼科镊两侧烧断输精管。
  7. 用眼科镊轻夹脂肪垫,将睾丸放回腹腔。
  8. 同上处理另一侧输精管。
  9. 缝合腹腔内部肌肉层和外部皮肤,单笼饲养。每天观察术后情况,连续3天。

致谢

感谢国家自然科学基金项目(Nos. 31572341, 31572347, 31772545, 31572348)、国家科技支撑计划项目(No. 2015BAI09B00)和“十三五”时期北京市属高校高水平教师队伍建设支持计划(IDHT20170516)的资助。

参考文献

  1. 王妍. (2019). 利用CRISPR/Cas9技术构建基因敲除长爪沙鼠动物模型. 首都医科大学硕士学位论文.
  2. Wang, Y., Zhao, P., Song, Z., Du, X., Huo, X., Lu, J., Liu, X., Lv, J., Li, C., Guo, M. and Chen, Z. (2020). Generation of gene-knockout Mongolian gerbils via CRISPR/Cas9 system. Front Bioeng Biotechnol 8: 780.
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Copyright: © 2021 The Authors; exclusive licensee Bio-protocol LLC.
引用格式:李长龙, 张静, 郭萌, 刘世景, 侯金莲, 朱艳敏, 吕珊, 杜小燕, 常在, 陈振文. (2021). 长爪沙鼠输精管结扎术. Bio-101: e1010931. DOI: 10.21769/BioProtoc.1010931.
How to cite: Li, C. L., Zhang, J., Guo, M., Lui, S. J. Hou, J., L, Zhu, Y., M., Lv, S., Du, X., Y., Chang, Z., and Chen, Z., W. (2021). Vasoligation in Mongolian Gerbils. Bio-101: e1010931. DOI: 10.21769/BioProtoc.1010931.
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