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Gfi1 小鼠出现早期进行性听力损失,并在内耳众多非毛细胞中诱导重组。

Gfi1 mice have early onset progressive hearing loss and induce recombination in numerous inner ear non-hair cells.

机构信息

Department of Otorhinolaryngology Head and Neck Surgery, University of Maryland School of Medicine, Baltimore, MD 21201, USA.

Department of Special Education and Communication Disorders, University of Nebraska Lincoln, Lincoln, Nebraska 68583-0738, USA.

出版信息

Sci Rep. 2017 Feb 9;7:42079. doi: 10.1038/srep42079.

Abstract

Studies of developmental and functional biology largely rely on conditional expression of genes in a cell type-specific manner. Therefore, the importance of specificity and lack of inherent phenotypes for Cre-driver animals cannot be overemphasized. The Gfi1 mouse is commonly used for conditional hair cell-specific gene deletion/reporter gene activation in the inner ear. Here, using immunofluorescence and flow cytometry, we show that the Gfi1 mice produce a pattern of recombination that is not strictly limited to hair cells within the inner ear. We observe a broad expression of Cre recombinase in the Gfi1 mouse neonatal inner ear, primarily in inner ear resident macrophages, which outnumber the hair cells. We further show that heterozygous Gfi1 mice exhibit an early onset progressive hearing loss as compared with their wild-type littermates. Importantly, vestibular function remains intact in heterozygotes up to 10 months, the latest time point tested. Finally, we detect minor, but statistically significant, changes in expression of hair cell-enriched transcripts in the Gfi1 heterozygous mice cochleae compared with their wild-type littermate controls. Given the broad use of the Gfi1 mice, both for gene deletion and reporter gene activation, these data are significant and necessary for proper planning and interpretation of experiments.

摘要

发育和功能生物学的研究在很大程度上依赖于以细胞类型特异性的方式对基因进行条件表达。因此,Cre 驱动动物的特异性和缺乏内在表型的重要性怎么强调都不为过。Gfi1 小鼠常用于在内耳中进行条件性毛细胞特异性基因缺失/报告基因激活。在这里,我们通过免疫荧光和流式细胞术显示,Gfi1 小鼠的重组模式并不严格局限于内耳中的毛细胞。我们在内耳新生 Gfi1 小鼠中观察到 Cre 重组酶的广泛表达,主要在内耳常驻巨噬细胞中,其数量超过毛细胞。我们进一步表明,与野生型同窝仔相比,杂合 Gfi1 小鼠表现出早期发作的进行性听力损失。重要的是,在测试的最晚时间点 10 个月时,杂合子的前庭功能仍然完整。最后,我们在 Gfi1 杂合子小鼠耳蜗中检测到毛细胞丰富的转录本表达的微小但具有统计学意义的变化,与野生型同窝仔对照相比。鉴于 Gfi1 小鼠广泛用于基因缺失和报告基因激活,这些数据对于实验的正确规划和解释具有重要意义和必要性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ca18/5299610/b11759f6556f/srep42079-f1.jpg

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