Page 21 - 南京医科大学学报自然科学版

第43卷第8期 南京医科大学学报（自然科学版）
2023年8月 Journal of Nanjing Medical University（Natural Sciences） ·1055 ·

·基础研究·

Pumilio家族基因诱导性敲除小鼠模型的构建

梁海波，邓晓凤，臧 敏，赵婷婷 *
南京医科大学生殖医学与子代健康全国重点实验室，江苏 南京 211166

［摘 要］ 目的：为探究出生后Pumilio家族的功能及其缺失对个体的影响，构建Pumilio1（Pum1）和Pumilio2（Pum2）双基因诱
2-/-
T2
导性敲除的小鼠模型。方法：通过小鼠交配实验得到 R26⁃ER Cre/+Pum1 flox/flox Pum 小鼠，并将同窝雄鼠设为对照组和实验
组。分别对实验组3周和成年小鼠注射他莫昔芬，在DNA、RNA和蛋白水平表征主要脏器中Pum1的敲除效果；并表征精子发
生过程中的病理变化、细胞增殖及凋亡情况。结果：实验组3周和成年雄鼠的胸腺和睾丸组织中Pum1表达水平极低，敲除效
率均高于50%，Pum1和Pum2双基因诱导性敲除小鼠模型构建成功。在注射他莫昔芬结束的第21天，3周雄鼠胸腺、脾脏和睾
丸的重量明显下降，睾丸病理HE染色实验显示精子形成障碍，TUNEL和BrdU染色后计数结果显示睾丸中生精细胞凋亡增加、
增殖减慢。成年小鼠在注射他莫昔芬恢复后，Pum1在胸腺、心、肝、睾丸中的表达仍有降低，但睾丸病理HE染色显示精子发生未
见异常。结论：该模型的建立首次展示Pumilio家族蛋白作为未来药物靶标的潜能，为深入研究Pumilio的功能及作为肿瘤治疗靶
蛋白的研究奠定了理论基础。
［关键词］ Pumilio；诱导性敲除；他莫昔芬；生殖；小鼠
［中图分类号］ Q78 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）08⁃1055⁃06
doi：10.7655/NYDXBNS20230803

An inducible knookout mouse model for Pumilio gene familiy

*
LIANG Haibo，DENG Xiaofeng，ZANG Min，ZHAO Tingting
State Key Laboratory of Reproductive Medicine and Offspring Heaith，Nanjing Medical University，Nanjing 211166，
China

［Abstract］ Objective： To determine the roles of Pumilio family in the postnatal development，we established the Pumilio1/Pumilio2
（Pum1 and Pum2）inducible knockout mouse model. Methods：Pum1 flox/flox Pum 2-/- mice were mated with R26⁃ERT2Cre/Cre mice to
obtain R26⁃ERT2Cre/+Pum1 flox/flox Pum 2-/- mice. The experiment group（three week old mice and adult mice）was injected with tamoxifen，
while the control group was injected with the same amount of peanut oil. The knockout efficiency of Pum1 at DNA，RNA and protein
levels of multiple organs in mice was determined. The histology of the testis as well as cell proliferation and apoptosis of the testicular
cells were examined. Results：The Pumilio1 RNA and protein of thymus and testis were knocked down significantly. The knock down
efficiency of Pum1 varies from tissues to tissues but reaching a minimum of 50%. The mouse model of Pum1 and Pum2 gene induced
knockout was successfully constructed. The weight of thymus，spleen and testis of male mice in the 3⁃week experiment group decreased
significantly，supporting the critical role of PUM in postnatal organ growth. The hematoxylin and eosin（HE），TUNEL and BrdU
staining of testis pathology showed disrupted spermatogenesis，increased apoptosis and decreased proliferation of spermatogenic cells
in 3 ⁃ week experiment group testis. Although，the Pum1 of each organ in adult group was knocked down at different levels，the
spermatogenesis of adult experimental group was not significantly affected. Conclusion：This inducible Pumilio knockout model
provides a new tool to study the roles of Pumilio in postnatal and adult mice，and supports the future development of Pumilio as
potential drug targets for diseases involving Pumilio⁃mediated translational control.
［Key words］ Pumilio；inducible knockout；tamoxifen；reproductive；mice
［J Nanjing Med Univ，2023，43（08）：1055⁃1060］

［基金项目］ 国家自然科学基金面上项目（31970792）
∗
通信作者（Corresponding author），E⁃mail： zhaotiti@njmu.edu.cn