Phospholipid scramblase isoform expression in pregnant rat uterus.
Phospholipid scramblase isoform expression in pregnant rat uterus.
复制标题
怀孕大鼠子宫中磷脂扰乱酶亚型的表达。
DOI：
10.1016/j.jsgi.2006.06.002
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发表时间：
2006
期刊：
影响因子：
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通讯作者：
Chien,EdwardK
中科院分区：
文献类型：
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作者：
Phillippe,Mark;Bradley,DianaF;Ji,Huiling;Oppenheimer,KarenH;Chien,EdwardK
目的：磷脂乱序酶（PLSCRs）是一类新的膜蛋白家族，能促进氨基磷脂从细胞膜内叶向细胞膜外叶的转运。在小鼠和人中已经报道了PLSCR的四种同种型（PLSCR 1 -4）。本报告中描述的研究试图表征子宫表达的PLSCR亚型在近期妊娠大鼠。方法：子宫组织从定时妊娠的Sprague-Dawley大鼠。分离总RNA，用DNase处理，并用于利用PLCSR同种型PCR引物的定性和定量逆转录酶-聚合酶链反应（RT-PCR）研究。大鼠脾cDNA噬菌体文库用作基于PCR的测序的模板，以确定大鼠中表达的PLSCR 3和PLSCR 4同源物的cDNA和翻译的氨基酸序列。结果：RT-PCR结果证实PLSCR 3和PLSCR 4在妊娠大鼠子宫内膜和肌层中有表达，而PLSCR 1和PLSCR 2在子宫组织中无表达。发现大鼠PLSCR 3同源物的cDNA序列为1642个核苷酸，与小鼠具有92%的同一性，与人PLSCR 3具有80%的同一性。大鼠PLSCR 4同源基因的cDNA序列为1879个核苷酸，与小鼠PLSCR 4同源基因的同源性为89%，与人PLSCR 4同源基因的同源性为72%。结论：PLSCR 3和PLSCR 4在子宫内的表达提供了一种动态机制，通过这种机制可以调节氨基磷脂的转运，从而调节各种膜蛋白的活性，这些膜蛋白参与炎症和凝血相关事件。
Objective:Phospholipid scramblases (PLSCRs), a family of novel membrane proteins, facilitate the translocation of aminophospholipids from the inner to the exterior leaf of the cell membrane. Four isoforms of PLSCR (PLSCR1-4) have been reported in mouse and human. The studies described in this report sought to characterize the uterine expression of the PLSCR isoforms in the near-term pregnant rat.Methods:Uterine tissue was obtained from timed-pregnant Sprague-Dawley rats. Total RNA was isolated, treated with DNase, and used in qualitative and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) studies utilizing PLCSR isoform PCR primers. A rat spleen cDNA bacteriophage library was used as a template for PCR-based sequencing to determine the cDNA and translated amino acid sequences for the PLSCR3 and PLSCR4 homologs expressed in rats. The 5′ and 3′ untranslated regions were obtained using 5′ and 3′ Rapid Amplification of cDNA Ends (RACE) techniques.Results:RT-PCR studies confirmed expression of PLSCR3 and PLSCR4 in the endometrial and myometrial layers of the pregnant rat uterus; in contrast, PLSCR1 and PLSCR2 were not found in uterine tissues. The cDNA sequence for the rat PLSCR3 homolog was found to be 1642 nucleotides, having 92% identity with mouse and 80% with human PLSCR3. The rat PLSCR4 homolog has a cDNA sequence of 1879 nucleotides, having an 89% identity with mouse and 72% identity with human PLSCR4 homologs.Conclusion:The intrauterine expression of PLSCR3 and PLSCR4 provides a dynamic mechanism by which aminophospholipid translocation can be regulated, thereby modulating the activity of various membrane proteins that are involved in inflammation and coagulation-related events.