Analysis of blood stem cell activity and cystatin gene expression in a mouse model presenting a chromosomal deletion encompassing Csta and Stfa2l1.
Analysis of blood stem cell activity and cystatin gene expression in a mouse model presenting a chromosomal deletion encompassing Csta and Stfa2l1.
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DOI：
10.1371/journal.pone.0007500
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发表时间：
2009-10-19
期刊：
影响因子：
3.7
通讯作者：
Sauvageau G
中科院分区：
文献类型：
--
作者：
Bilodeau M;MacRae T;Gaboury L;Laverdure JP;Hardy MP;Mayotte N;Paradis V;Harton S;Perreault C;Sauvageau G
半胱氨酸蛋白酶抑制剂蛋白超家族的特征在于存在显示半胱氨酸蛋白酶抑制活性的保守序列（例如，朝向组织蛋白酶）。1型和2型半胱氨酸蛋白酶抑制剂由25个基因编码，其中23个基因被分组为位于小鼠染色体16和2上的2个簇。这些基因中的大多数在小鼠发育和造血中的表达和重要作用仍然很差。在这项研究中，我们描述了一套定量实时PCR检测和全球正常小鼠组织中的胱抑素基因的表达谱。受益于我们收集的携带大染色体缺失的DelES胚胎干细胞克隆（将在其他地方报道），我们选择了一个16号染色体95 kb区域缺失的克隆（Del 16 qB 3 Δ/+）。在该特定克隆中，2个胱抑素基因，即Csta和Stfa 2l 1，以及2个其它基因（Fam 162 a，Ccdc 58）和相关的基因间区域缺失沿着。在此基础上，我们建立了一种新的纯合子突变小鼠模型（Del 16 qB 3 Δ/16 qB 3 Δ），以评估2种缺失的半胱氨酸蛋白酶抑制剂的体内生物学功能。Stfa 2l 1基因在野生型胎儿肝脏、骨髓和脾脏中表达较高，而Csta则普遍表达。纯合子Del 16 qB 3 Δ/16 qB 3 Δ动物表型正常，可生育，对自发或辐射诱导的肿瘤形成不明显敏感。这些突变小鼠的造血干细胞和祖细胞活性也正常。有趣的是，定量实时PCR表达谱显示Del 16 qB 3 Δ/16 qB 3 Δ造血组织中Stfa 2l 1/Csta造血相关基因（Stfa 1、Stfa 2和Stfa 3）的表达水平显著增加，表明这些候选基因可能有助于代偿机制。总体而言，本研究提出了一种优化的方法来全球监测胱抑素基因的表达，以及一个新的小鼠模型缺乏Stfa 2l 1/Csta基因，扩大了现有的工具，解剖胱抑素在正常和病理条件下的作用。
The cystatin protein superfamily is characterized by the presence of conserved sequences that display cysteine protease inhibitory activity (e.g., towards cathepsins). Type 1 and 2 cystatins are encoded by 25 genes of which 23 are grouped in 2 clusters localized on mouse chromosomes 16 and 2. The expression and essential roles of most of these genes in mouse development and hematopoiesis remain poorly characterized. In this study, we describe a set of quantitative real-time PCR assays and a global expression profile of cystatin genes in normal mouse tissues. Benefiting from our collection of DelES embryonic stem cell clones harboring large chromosomal deletions (to be reported elsewhere), we selected a clone in which a 95-kb region of chromosome 16 is missing (Del16qB3Δ/+). In this particular clone, 2 cystatin genes, namely Csta and Stfa2l1 are absent along with 2 other genes (Fam162a, Ccdc58) and associated intergenic regions. From this line, we established a new homozygous mutant mouse model (Del16qB3Δ/16qB3Δ) to assess the in vivo biological functions of the 2 deleted cystatins. Stfa2l1 gene expression is high in wild-type fetal liver, bone marrow, and spleen, while Csta is ubiquitously expressed. Homozygous Del16qB3Δ/16qB3Δ animals are phenotypically normal, fertile, and not overtly susceptible to spontaneous or irradiation-induced tumor formation. The hematopoietic stem and progenitor cell activity in these mutant mice are also normal. Interestingly, quantitative real-time PCR expression profiling reveals a marked increase in the expression levels of Stfa2l1/Csta phylogenetically-related genes (Stfa1, Stfa2, and Stfa3) in Del16qB3Δ/16qB3Δ hematopoietic tissues, suggesting that these candidate genes might be contributing to compensatory mechanisms. Overall, this study presents an optimized approach to globally monitor cystatin gene expression as well as a new mouse model deficient in Stfa2l1/Csta genes, expanding the available tools to dissect cystatin roles under normal and pathological conditions.
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影响因子：
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