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发育中星形胶质细胞初级纤毛的体内基因标记

In vivo genetic labeling of primary cilia in developing astrocytes.

作者信息

Bear Rachel, Wei Claire, Caspary Tamara

机构信息

Department of Human Genetics, Emory University School of Medicine, 615 Michael Street Suite 301, Atlanta, GA, 30322, USA.

Emory Graduate Program in Neuroscience, Emory University, Atlanta, GA, 30322, USA.

出版信息

BMC Mol Cell Biol. 2025 Jul 24;26(1):23. doi: 10.1186/s12860-025-00547-7.

Abstract

BACKGROUND

Astrocyte cilia are largely understudied due to the lack of available tools. Astrocyte research advanced with the establishment of Aldh1l1-Cre, an inducible Cre line that specifically targets the astrocyte lineage. Here, we develop and compare genetic models that label astrocyte cilia in the developing prefrontal cortex (PFC) using Aldh1l1-Cre and Cre-dependent cilia reporters. We evaluate these models by testing different tamoxifen-induction protocols and quantifying the percentage of astrocytes labeled with the cilia reporters.

RESULTS

We show that tamoxifen dosage impacts the expression of cilia reporters in astrocytes. We uncover the maximum cilia-labeling efficiency using recombined, constitutively-expressed cilia reporters.

CONCLUSION

The data reveal that only a subset of SOX9- positive astrocytes in the PFC possess cilia throughout development. Our work highlights the utility of Cre-Lox systems to target specific cell types and the importance of carefully validating genetic models.

摘要

背景

由于缺乏可用的工具,星形胶质细胞的纤毛在很大程度上未得到充分研究。随着Aldh1l1-Cre的建立,星形胶质细胞的研究取得了进展,Aldh1l1-Cre是一种可诱导的Cre系,专门针对星形胶质细胞谱系。在这里,我们开发并比较了使用Aldh1l1-Cre和Cre依赖性纤毛报告基因标记发育中的前额叶皮质(PFC)中星形胶质细胞纤毛的遗传模型。我们通过测试不同的他莫昔芬诱导方案并量化用纤毛报告基因标记的星形胶质细胞的百分比来评估这些模型。

结果

我们表明他莫昔芬剂量会影响星形胶质细胞中纤毛报告基因的表达。我们使用重组的、组成型表达的纤毛报告基因揭示了最大的纤毛标记效率。

结论

数据表明,在整个发育过程中,PFC中只有一部分SOX9阳性星形胶质细胞具有纤毛。我们的工作强调了Cre-Lox系统针对特定细胞类型的实用性以及仔细验证遗传模型的重要性。

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