Non-invasive visualization of the lipid product of class I PI3K in transgenic mouse models.
Non-invasive visualization of the lipid product of class I PI3K in transgenic mouse models.
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DOI：
10.1042/bst0350215
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发表时间：
2007-04
影响因子：
3.9
通讯作者：
T. Sasaki;J. Sasaki;K. Watanabe;A. Suzuki
中科院分区：
文献类型：
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作者：
T. Sasaki;J. Sasaki;K. Watanabe;A. Suzuki
磷脂酰肌醇3激酶（phosphoinositide 3-kinases，PI 3 Ks）通过产生3，4，5-三磷酸磷脂酰肌醇（phosphatidylinositol 3，4，5-trisphosphate，PI（3，4，5）P（3））调节许多重要的细胞反应。为了促进PI（3，4，5）P（3）在活的原代细胞中的时空表征，我们产生了表达PI（3，4，5）P（3）/PI（3，4）P（2）（磷脂酰肌醇3，4-二磷酸）的荧光生物探针的转基因小鼠[AktPH（Akt pleckstrin同源结构域）-GFP（绿色荧光蛋白）Tg（转基因）小鼠]的新品系。通过将AktPH-GFP Tg小鼠与缺乏特定磷酸肌醇代谢酶的基因靶向“敲除”小鼠品系杂交，我们已经能够评估每种酶对PI（3，4，5）P（3）在迁移中性粒细胞中定位的贡献。我们的研究结果表明，在fMet-Leu-Phe（N-甲酰甲硫氨酰-亮氨酰苯丙氨酸;“趋化肽”）刺激的中性粒细胞迁移过程中，PI 3 K γ和PI（3，4，5）P（3）磷酸酶SHIP 1 [含SH 2（Src同源2）的肌醇磷酸酶-1]是PI（3，4，5）P（3）动力学的关键调节因子。我们的研究也验证了荧光转基因策略在生理和病理情况下研究PI（3，4，5）P（3）代谢。
PI3Ks (phosphoinositide 3-kinases) regulate many critical cellular responses by producing PI(3,4,5)P(3) (phosphatidylinositol 3,4,5-trisphosphate). To facilitate the spatio-temporal characterization of PI(3,4,5)P(3) in living primary cells, we generated a novel strain of transgenic mice [AktPH (Akt pleckstrin homology domain)-GFP (green fluorescent protein) Tg (transgenic) mice] that express a fluorescent bioprobe for PI(3,4,5)P(3)/PI(3,4)P(2) (phosphatidylinositol 3,4-bisphosphate). By crossing AktPH-GFP Tg mice with strains of gene-targeted 'knockout' mice lacking a particular phosphoinositide-metabolizing enzyme, we have been able to evaluate the contribution of each enzyme to PI(3,4,5)P(3) localization in migrating neutrophils. Our results indicate that PI3Kgamma and the PI(3,4,5)P(3) phosphatase SHIP1 [SH2 (Src homology 2)-containing inositol phosphatase-1] are the key regulators of PI(3,4,5)P(3) dynamics during fMet-Leu-Phe (N-formylmethionyl-leucylphenylalanine; 'chemotactic peptide')-stimulated neutrophil migration. Our study has also validated the fluorescent transgenic strategy for studying PI(3,4,5)P(3) metabolism in physiological and pathological situations.