Membrane Dynamics Induced by a Phosphatidylinositol 3,4,5-Trisphosphate Optogenetic Tool

• Published in: Analytical Sciences Vol. 35; no. 1; pp. 57 - 63
• Main Authors: UEDA, Yoshibumi, II, Tatsuhito, AONO, Yuki, SUGIMOTO, Naotoshi, SHINJI, Seiichi, YOSHIDA, Hiroshi, SATO, Moritoshi
• Format: Journal Article
• Language: English
• Published: Singapore The Japan Society for Analytical Chemistry 10.01.2019; Springer Nature Singapore; Japan Science and Technology Agency
• Subjects: Actin; Actins - metabolism; Analytical Chemistry; Animals; Cancer; Cell Membrane - metabolism; Cell Movement; Cellular structure; Cercopithecus aethiops; Chemistry; COS Cells; cryptochrome 2; Depolymerization; Dynamic structural analysis; Dynamics; Filopodia; HEK293 Cells; Humans; Investigations; Lamellipodia; Lipid; Lipids; Metastases; Mice; Microscopy; Microscopy, Confocal; Motility; NIH 3T3 Cells; optogenetics; Optogenetics - methods; Phagocytosis; Pharmacology; Phosphatidylinositol; Phosphatidylinositol 3,4,5-triphosphate; phosphatidylinositol 3,4,5-trisphosphate; Phosphatidylinositol Phosphates - metabolism; PIP3; Protein Binding; Sensitivity and Specificity; Spatial resolution; structured illumination microscopy; Wound healing; phosphatidylinositol 3,4,5-trisphosphate; optogenetics; cryptochrome 2; lamellipodia; PIP3; structured illumination microscopy Acknowledgements This work was supported by CREST, JST (to M. S.) and PRIME from Japan Agency for Medical Research and Development (to Y. U.); Lipid; structured illumination microscopy
• ISSN: 0910-6340; 1348-2246
• DOI: 10.2116/analsci.18SDP06

Membrane dynamic structures such as filopodia, lamellipodia, and ruffles have important cellular functions in phagocytosis and cell motility, and in pathological states, such as cancer metastasis. Phosphatidylinositol 3,4,5-trisphosphate (PIP3) is a crucial lipid that regulates PIP3 dynamics. Investigations of how PIP3 is involved in these functions have mainly relied on pharmacological interventions, and therefore have not generated detailed spatiotemporal information concerning membrane dynamics upon PIP3 production. In the present study, we applied an optogenetic approach using the CRY2–CIBN system. Using this system, we revealed that local PIP3 generation induced directional cell motility and membrane ruffles in COS7 cells. Furthermore, combined with structured illumination microscopy (SIM), membrane dynamics were investigated with high spatial resolution. We observed PIP3-induced apical ruffles and unique actin fiber behavior in that a single actin fiber protruded from the plasma membrane was taken up into the plasma membrane without depolymerization. This system has the potential to investigate other high-level cell motility and dynamic behaviors, such as cancer cell invasion and wound healing with high spatiotemporal resolution, and could provide new insights of biological sciences for membrane dynamics.