Normal spermatogenesis in Fank1 (fibronectin type 3 and ankyrin repeat domains 1) mutant mice
Normal spermatogenesis in Fank1 (fibronectin type 3 and ankyrin repeat domains 1) mutant mice
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Fank1（纤连蛋白 3 型和锚蛋白重复结构域 1）突变小鼠的正常精子发生
DOI：
10.7717/peerj.6827
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发表时间：
2019-04-24
期刊：
影响因子：
2.7
通讯作者：
Liu, Mingxi
中科院分区：
文献类型：
--
作者：
Zhang, Jintao;Zhang, Xin;Liu, Mingxi
研究背景Fank 1基因是脊椎动物中一个古老的、进化上保守的基因。基于短发夹RNA（shRNA）的基因敲减转基因小鼠具有由凋亡生殖细胞增加引起的少精子症。在这项研究中，我们研究了Fank 1的体内功能。方法在本研究中，我们使用CRISPR/Cas9系统产生Fank 1基因敲除小鼠。然后，我们研究了Fank 1的表型和体内功能。睾丸和附睾组织进行组织学和免疫荧光染色。在末端脱氧核苷酸转移酶dUTP缺口末端标记测定中分析凋亡细胞。还评价了生育力和精子计数。利用GTEx数据库进行基因表达数量性状位点的检测，并采用定量RT-PCR方法分析候选基因和邻近单核苷酸多态性基因的mRNA表达。结果与Fank 1基因敲除模型相比，Fank 1 −/−纯合子模型的附睾精子含量和凋亡细胞数无明显变化。此外，Dusp 1，Klk 1b 21和Klk 1b 27 mRNA的表达在Fank 1基因敲除的睾丸中检测到不同的模式。这些结果揭示了Fank 1基因敲除小鼠与Fank 1基因敲除小鼠在分子水平上的差异，为群体遗传学研究提供了基础资料。
Background The fibronectin type 3 and ankyrin repeat domains 1 gene, Fank1, is an ancient, evolutionarily conserved gene present in vertebrates. Short-hairpin RNA (shRNA)-based knockdown transgenic mice have oligospermia caused by an increase in apoptotic germ cells. In this study, we investigated the in vivo function of Fank1. Methods In this study, we generated Fank1-knockout mice using the CRISPR/Cas9 system. We then investigated the phenotype and in vivo function of Fank1. Testes and epididymis tissues were analyzed by histological and immunofluorescence staining. Apoptotic cells were analyzed in terminal deoxynucleotidyl transferase dUTP nick end-labeling assays. Fertility and sperm counts were also evaluated. The GTEx database were used to assess gene expression quantitative trait loci and mRNA expression of candidate genes and genes neighboring single nucleotide polymorphisms was analyzed by quantitative RT-PCR. Results In contrast to the Fank1-knockdown model, no significant changes in epididymal sperm content and the number of apoptotic cells were observed in Fank1−/− homozygotes. In addition, a different pattern of Dusp1, Klk1b21 and Klk1b27 mRNA expression was detected in Fank1-knockout testis. These results reveal differences in the molecular changes between Fank1-knockdown mice and Fank1-knockout mice and provide a basic resource for population genetics studies.