Department of Biochemistry, University of Iowa, Iowa City, IA 52242, USA.
Department of Ophthalmology & Visual Sciences and Institute for Vision Research, University of Iowa, Iowa City, IA 52242, USA.
Exp Eye Res. 2018 May;170:108-116. doi: 10.1016/j.exer.2018.02.022. Epub 2018 Feb 24.
The 14-3-3 family of proteins has undergone considerable expansion in higher eukaryotes with humans and mice expressing seven isoforms (β, ε, η, γ, θ, ζ, and σ) from seven distinct genes (YWHAB, YWAHE, YWHAH, YWHAG, YWHAQ, YWHAZ, and SFN). Growing evidence indicates that while highly conserved, these isoforms are not entirely functionally redundant as they exhibit unique tissue expression profiles, subcellular localization, and biochemical functions. A key limitation in our understanding of 14-3-3 biology lies in our limited knowledge of cell-type specific 14-3-3 expression. Here we provide a characterization of 14-3-3 expression in whole retina and isolated rod photoreceptors using reverse-transcriptase digital droplet PCR. We find that all 14-3-3 genes with the exception of SFN are expressed in mouse retina with YWHAQ and YWHAE being the most highly expressed. Rod photoreceptors are enriched in YWHAE (14-3-3 ε). Immunohistochemistry revealed that 14-3-3 ε and 14-3-3 ζ exhibit unique distributions in photoreceptors with 14-3-3 ε restricted to the inner segment and 14-3-3 ζ localized to the outer segment. Our data demonstrates that, in the retina, 14-3-3 isoforms likely serve specific functions as they exhibit unique expression levels and cell-type specificity. As such, future investigations into 14-3-3 function in rod photoreceptors should be centered on 14-3-3 ε and 14-3-3 ζ, depending on the subcellular region of question.
蛋白质 14-3-3 家族在高等真核生物中经历了相当大的扩张,人类和小鼠分别从七个不同的基因(YWHAB、YWAHE、YWHAH、YWHAG、YWHAQ、YWHAZ 和 SFN)表达七种同工型(β、ε、η、γ、θ、ζ 和 σ)。越来越多的证据表明,尽管这些同工型高度保守,但它们并非完全具有功能冗余性,因为它们表现出独特的组织表达谱、亚细胞定位和生化功能。我们对 14-3-3 生物学的理解的一个关键限制在于我们对细胞类型特异性 14-3-3 表达的了解有限。在这里,我们使用逆转录数字液滴 PCR 对整个视网膜和分离的杆状光感受器中的 14-3-3 表达进行了描述。我们发现除了 SFN 之外,所有 14-3-3 基因在小鼠视网膜中均有表达,其中 YWHAQ 和 YWHAE 的表达水平最高。杆状光感受器富含 YWHAE(14-3-3ε)。免疫组织化学显示,14-3-3ε和 14-3-3ζ在光感受器中表现出独特的分布,14-3-3ε局限于内节,14-3-3ζ定位于外节。我们的数据表明,在视网膜中,14-3-3 同工型可能具有特定的功能,因为它们表现出独特的表达水平和细胞类型特异性。因此,未来对杆状光感受器中 14-3-3 功能的研究应集中在 14-3-3ε和 14-3-3ζ上,具体取决于所研究的亚细胞区域。