The spatial patterning mechanisms of cytoskeletons during cancer cell invasion analyzed by 3D multitarget super-resolution imaging

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K07384
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cell biology
• Research Institution: Kyoto University
• Principal Investigator: Kiuchi Tai 京都大学, 医学研究科, 准教授 (70443984)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 超解像顕微鏡法 / アクチン / 細胞骨格

Outline of Final Research Achievements

Cancer cells rearrange the cytoskeletons and focal adhesions in appropriate 3D patterns through interactions with the surroundings for cancer invasion. To study the 3D rearrangement mechanism of the cytoskeletons, multitargets super-resolution imaging IRIS was developed into 3D imaging. For the 3D imaging, HILO illumination and adaptive optics were inserted into the optical path of the microscope. To reduce the background light, the Z resolution and Z drift, the excitation conditions were examined in detail. Then, the 3D networks of actin filaments and microtubules in the apical region of cells were visualized. Furthermore, it was found that EGF receptors, which induce the rearrangement of cytoskeletons, are accumulated in the interspace between actin stress fibers by EGF stimulation and then are endocytosed.

Free Research Field

細胞生物学

Academic Significance and Societal Importance of the Research Achievements

これまで超解像顕微鏡法を３次元化するために様々な取り組みが行われてきた。しかし、蛍光退色や背景光、Z分解能、測定中のZドリフトなど励起条件や測定条件で検討する課題が多い。さらに多重染色超解像イメージングは、標的の標識率や複数の蛍光色素を用いることによる色収差や球面収差の問題が追加される。IRISは、結合解離プローブを用いることで、標識率の上限を突破し、さらに同一の蛍光色素を結合解離プローブに接合することで、この色収差や球面収差の問題を回避している。IRISを３次元イメージングへと発展させることで、今後、細胞や組織の３次元的な構造の詳細が明らかになることが期待できる。