Microarray analysis of prothrombin knockdown in zebrafish.
Microarray analysis of prothrombin knockdown in zebrafish.
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DOI：
10.1016/j.bcmd.2009.04.001
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发表时间：
2009-09
影响因子：
2.3
通讯作者：
Jagadeeswaran, Pudur
中科院分区：
文献类型：
--
作者：
Day, Kenneth R.;Jagadeeswaran, Pudur
丝氨酸蛋白酶凝血酶在凝血级联反应中由其前体凝血酶原产生，并在通过蛋白酶激活受体介导的纤维蛋白沉积和血小板活化中发挥核心作用。斑马鱼中凝血酶原的敲除先前显示重现了在凝血酶原敲除小鼠中观察到的表型，例如不存在血液心包水肿和出血。然而，凝血酶在胚胎发生过程中的作用尚未完全了解。为了找到在血液循环之前胚胎发生中受潜在凝血酶信号传导影响的基因，使用从反义注射的、敲低的胚胎与在受精后20小时注射的错配胚胎制备的总RNA进行微阵列分析。共63个上调和下调的基因进行了鉴定与重复微阵列使用染料逆转和两倍的差异限制。对筛选出的10个基因进行真实的实时RT-PCR检测，证实了这些基因的表达变化。一个特定的基因，phlda 3，至少有11倍上调，原位杂交显示，在敲除胚胎的中枢神经系统，鳃弓，头部内胚层的phlda 3表达的扩展。这些基因的鉴定凝血酶调节根据微阵列分析应该提供一个更好的理解凝血酶活性的影响，在早期脊椎动物胚胎。
The serine protease thrombin is generated from its precursor, prothrombin, in the coagulation cascade and plays a central role in fibrin deposition and platelet activation mediated through the protease activated receptors. Knockdown of prothrombin in the zebrafish was previously shown to recapitulate the phenotype observed in prothrombin knockout mice, such as an absence of blood pericardial edema, and hemorrhage. However, the role of thrombin during embryogenesis is not fully understood. To find genes affected by potential thrombin signaling in embryogenesis before blood circulation microarray analysis was performed using total RNA prepared from antisense-injected, knockdown embryos versus mismatch-injected at 20 hours post fertilization. A total of 63 upregulated and downregulated genes were identified with duplicate microarrays using dye reversal and a two-fold difference limitation. Real time RT-PCR for 10 selected genes identified by the microarray confirmed the expression changes in these genes. One particular gene, phlda3, was at least eleven fold upregulated, and in situ hybridization revealed expansion of phlda3 expression in the central nervous system, branchial arches, and head endoderm in knockdown embryos. The identification of these genes regulated by thrombin according to microarray analysis should provide a greater understanding of the effects of thrombin activity in the early vertebrate embryo.
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影响因子：
16.2
作者：
SUIDAN, HS;STONE, SR;MONARD, D
通讯作者：
MONARD, D
影响因子：
2.6
作者：
Argenton, F;Giudici, S;Beltrame, M
通讯作者：
Beltrame, M
DOI：
10.1016/s0167-4889(96)00109-7
发表时间：
1996-12-12
影响因子：
5.1
作者：
Abrams, CS;Zhao, W;Brass, LF
通讯作者：
Brass, LF
影响因子：
2.4
作者：
De Martino, SP;Errington, F;Austin, CA
通讯作者：
Austin, CA
DOI：
10.1073/pnas.85.10.3440
发表时间：
1988-05-01
影响因子：
11.1
作者：
GURWITZ, D;CUNNINGHAM, DD
通讯作者：
CUNNINGHAM, DD