PI3Ks (phosphoinositide 3-kinases) regulate many critical cellular responses by producing PI(3,4,5)P3 (phosphatidylinositol 3,4,5-trisphosphate). To facilitate the spatio-temporal characterization of PI(3,4,5)P3 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)P3/PI(3,4)P2 (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)P3 localization in migrating neutrophils. Our results indicate that PI3Kγ and the PI(3,4,5)P3 phosphatase SHIP1 [SH2 (Src homology 2)-containing inositol phosphatase-1] are the key regulators of PI(3,4,5)P3 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)P3 metabolism in physiological and pathological situations.
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Conference Article| March 20 2007
Non-invasive visualization of the lipid product of class I PI3K in transgenic mouse models
T. Sasaki 1
*Department of Pathology and Immunology, Akita University School of Medicine, 1-1-1 Hondo, Akita 010-8543, Japan
†PRESTO (Precursory Research for Embryonic Science and Technology), Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012, Japan
1To whom correspondence should be addressed (email firstname.lastname@example.org).
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T. Sasaki, J. Sasaki, K. Watanabe, A. Suzuki; Non-invasive visualization of the lipid product of class I PI3K in transgenic mouse models. Biochem Soc Trans 1 April 2007; 35 (2): 215–218. doi: https://doi.org/10.1042/BST0350215
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