Purification of GFRα1+ and GFRα1- Spermatogonial Stem Cells Reveals a Niche-Dependent Mechanism for Fate Determination.
Purification of GFRα1+ and GFRα1- Spermatogonial Stem Cells Reveals a Niche-Dependent Mechanism for Fate Determination.
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DOI：
10.1016/j.stemcr.2017.12.009
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发表时间：
2018-02-13
影响因子：
5.9
通讯作者：
Artandi SE
中科院分区：
文献类型：
--
作者：
Garbuzov A;Pech MF;Hasegawa K;Sukhwani M;Zhang RJ;Orwig KE;Artandi SE
未分化的精原细胞由干细胞和祖细胞组成，这些干细胞和祖细胞表现出标记物（包括细胞表面受体GFRα1）的异质性表达。分离GFRα1+与GFRα1-未分化精原细胞的技术挑战排除了这些亚群的比较分子表征及其作为干细胞的功能评价。在这里，我们开发了一种方法，通过荧光激活细胞分选纯化这些亚群，并显示GFRα1+和GFRα1-未分化精原细胞都表现出较高的移植活性，而不同的主要受体酪氨酸激酶信号和细胞周期。我们鉴定了细胞表面分子黑素细胞粘附分子（MCAM）在这些群体中的差异表达，并表明MCAM抗体允许从成年野生型小鼠中分离高度富集的GFRα1+和GFRα1-精原细胞群体。在生殖细胞培养中，GFRα1-细胞上调MCAM表达以响应胶质细胞系源性神经营养因子（GDNF）/成纤维细胞生长因子（FGF）刺激。在移植宿主中，GFRα1-精原细胞产生GFRα1+精原细胞并恢复精子发生，尽管其速率低于GFRα1+对应物。总之，这些数据为干细胞库模型提供了支持，其中GFRα1+和GFRα1-细胞密切相关，但显示出关键的细胞内在差异，并且可以部分基于获得生态位因子在两种状态之间相互转换。一种新的流式细胞仪纯化GFRα 1+和GFRα1-未分化精原细胞的方法GFR α 1+和GFRα1-细胞在受体酪氨酸激酶信号转导上的差异表面MCAM表达的差异可以区分GFRα1+和GFRα1-细胞GFRα 1-细胞具有GFRα1+细胞的三分之一的移植活性。Garbuzov及其同事设计了一种新的策略，基于端粒酶和GFRα1的表达，从TertTomato报告小鼠的成年睾丸中分离GFRα1+和GFRα1-未分化精原细胞的纯群体。转录谱显示GFRα1+和GFRα1-细胞之间具有显著的相似性，并且两个群体通过移植均显示出升高的干细胞活性。
Undifferentiated spermatogonia comprise a pool of stem cells and progenitor cells that show heterogeneous expression of markers, including the cell surface receptor GFRα1. Technical challenges in isolation of GFRα1+ versus GFRα1– undifferentiated spermatogonia have precluded the comparative molecular characterization of these subpopulations and their functional evaluation as stem cells. Here, we develop a method to purify these subpopulations by fluorescence-activated cell sorting and show that GFRα1+ and GFRα1– undifferentiated spermatogonia both demonstrate elevated transplantation activity, while differing principally in receptor tyrosine kinase signaling and cell cycle. We identify the cell surface molecule melanocyte cell adhesion molecule (MCAM) as differentially expressed in these populations and show that antibodies to MCAM allow isolation of highly enriched populations of GFRα1+ and GFRα1– spermatogonia from adult, wild-type mice. In germ cell culture, GFRα1– cells upregulate MCAM expression in response to glial cell line-derived neurotrophic factor (GDNF)/fibroblast growth factor (FGF) stimulation. In transplanted hosts, GFRα1– spermatogonia yield GFRα1+ spermatogonia and restore spermatogenesis, albeit at lower rates than their GFRα1+ counterparts. Together, these data provide support for a model of a stem cell pool in which the GFRα1+ and GFRα1– cells are closely related but show key cell-intrinsic differences and can interconvert between the two states based, in part, on access to niche factors. A new method to purify GFRα1+ and GFRα1– undifferentiated spermatogonia by FACS GFRα1+ and GFRα1– cells differ in receptor tyrosine kinase signaling Differential surface MCAM expression can distinguish GFRα1+ and GFRα1– cells GFRα1– cells have a third of the transplantation activity of GFRα1+ cells In this article, Garbuzov and colleagues devise a new strategy for isolating pure populations of GFRα1+ and GFRα1– undifferentiated spermatogonia from adult testis of TertTomato reporter mice based on expression of telomerase and GFRα1. Transcriptional profiling showed a remarkable similarity between GFRα1+ and GFRα1– cells, and both populations showed elevated stem cell activity by transplantation.
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影响因子：
5.2
作者：
Hasegawa, Kazuteru;Namekawa, Satoshi H.;Saga, Yumiko
通讯作者：
Saga, Yumiko
影响因子：
23.9
作者：
Hara, Kenshiro;Nakagawa, Toshinori;Enomoto, Hideki;Suzuki, Mikiko;Yamamoto, Masayuki;Simons, Benjamin D.;Yoshida, Shosei
通讯作者：
Yoshida, Shosei
影响因子：
3.6
作者：
Kanatsu-Shinohara, Mito;Morimoto, Hiroko;Shinohara, Takashi
通讯作者：
Shinohara, Takashi
影响因子：
--
作者：
KLUIN, PM;KRAMER, MF;DEROOIJ, DG
通讯作者：
DEROOIJ, DG
影响因子：
3.6
作者：
Buageaw, A;Sukhwani, M;Schlatt, S
通讯作者：
Schlatt, S