Sirtuin1 对于小鼠滋养层细胞的正常分化和胎盘发育是必需的。
Sirtuin1 is required for proper trophoblast differentiation and placental development in mice.
机构信息
Department of Pathology, University of California San Diego, La Jolla, CA, 92093, USA; Sanford Consortium for Regenerative Medicine, University of California San Diego, La Jolla, CA, 92093, USA.
Sanford Consortium for Regenerative Medicine, University of California San Diego, La Jolla, CA, 92093, USA; Department of Reproductive Medicine, University of California San Diego, La Jolla, CA, 92093, USA.
出版信息
Placenta. 2018 Feb;62:1-8. doi: 10.1016/j.placenta.2017.12.002. Epub 2017 Dec 7.
INTRODUCTION
Placental insufficiency, arising from abnormal trophoblast differentiation and function, is a major cause of fetal growth restriction. Sirtuin-1 (Sirt1) is a ubiquitously-expressed NAD-dependent protein deacetylase which plays a key role in numerous cellular processes, including cellular differentiation and metabolism. Though Sirt1 has been widely studied, its role in placentation and trophoblast differentiation is unclear.
METHOD
Sirt1-heterozygous mice were mated and evaluated at various points during embryogenesis. In situ hybridization and immunohistochemistry were used to further characterize the placental phenotype of Sirt1-null mice. Wild-type (WT) and Sirt1-null mouse trophoblast stem cell (TSC) lines were derived from e3.5 littermate blastocysts. These cells were then evaluated at various points following differentiation. Differentiation was evaluated by expression of lineage specific markers using qPCR and flow cytometry, as well as Matrigel invasion assays. Global gene expression changes were evaluated using microarray-based RNA profiling; changes in specific pathways were validated using qPCR and western blot.
RESULTS
In the absence of Sirt1, both embryos and placentas were small, with placentas showing abnormalities in both the labyrinthine layer and junctional zone. Sirt1-null TSCs exhibited an altered phenotype in both undifferentiated and differentiated states, phenotypes which corresponded to changes in pathways relevant to both TSC maintenance and differentiation. Specifically, Sirt1-null TSC showed blunted differentiation, and appeared to be suspended in an Epcam trophoblast progenitor state.
DISCUSSION
Our results suggest that Sirt1 is required for proper TSC differentiation and placental development.
简介
胎盘功能不全是由滋养细胞分化和功能异常引起的，是胎儿生长受限的主要原因。Sirtuin-1（Sirt1）是一种广泛表达的 NAD 依赖性蛋白去乙酰化酶，在许多细胞过程中发挥关键作用，包括细胞分化和代谢。尽管 Sirt1 已被广泛研究，但它在胎盘形成和滋养细胞分化中的作用尚不清楚。
方法
Sirt1 杂合子小鼠在胚胎发生的不同阶段进行交配和评估。原位杂交和免疫组织化学用于进一步表征 Sirt1 缺失小鼠的胎盘表型。从 e3.5 胎盘中获得野生型（WT）和 Sirt1 缺失小鼠滋养层干细胞（TSC）系。然后在分化后的不同时间点评估这些细胞。使用 qPCR 和流式细胞术以及 Matrigel 侵袭测定评估分化。使用基于微阵列的 RNA 谱分析评估整体基因表达变化；使用 qPCR 和 Western blot 验证特定途径的变化。
结果
在没有 Sirt1 的情况下，胚胎和胎盘都很小，胎盘在绒毛层和连接区都出现异常。在未分化和分化状态下，Sirt1 缺失的 TSC 均表现出异常表型，这些表型与与 TSC 维持和分化相关的特定途径的变化相对应。具体而言，Sirt1 缺失的 TSC 分化受到抑制，并且似乎处于 Epcam 滋养细胞祖细胞状态。
讨论
我们的结果表明 Sirt1 是 TSC 分化和胎盘发育所必需的。
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