A RhoA-FRET Biosensor Mouse for Intravital Imaging in Normal Tissue Homeostasis and Disease Contexts

Cell Reports

Shereen Kadir¹  Jennifer P. Morton¹  Juliane P. Schwarz¹  Agata Mrowinska¹  Douglas Strathdee¹  Ewan J. McGhee¹  David Stevenson¹  Owen J. Sansom¹  Michael S. Samuel²  Richard P. Harvey³  Gonzalo del Monte-Nieto³  Kurt I. Anderson⁴  Edna C. Hardeman⁵  Andrius Masedunskas⁵  Peter W. Gunning⁶  Anna-Karin E. Johnsson⁷  Heidi C.E. Welch⁷  Wilfred Leung⁸  Morghan C. Lucas⁸  Tatyana Chtanova⁸  Anaiis Zaratzian⁸  Nadine Reischmann⁸  Claire Vennin⁸  Astrid Magenau⁸  Christopher J. Ormandy⁸  Herbert Herzog⁸  Alice Boulghourjian⁸  Andrew M. Law⁸  Max Nobis⁸  Sean C. Warren⁸  James R.W. Conway⁸  Julian M.W. Quinn⁸  Jacqueline Bailey⁸  Paul Timpson⁸  Stacey N. Walters⁸  Lei Zhang⁸  David Gallego-Ortega⁸  Monica Killen⁸  David Herrmann⁸  Shane T. Grey⁸  Paul A. Baldock⁸  Marina Pajic⁸  Peter I. Croucher⁸ 

[1] Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G611BD, UK;Centre for Cancer Biology, SA Pathology and University of South Australia School of Medicine, University of Adelaide, Adelaide, SA 5000, Australia;Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research Institute, Sydney, NSW 2010, Australia;Francis Crick Institute, London NW11AT, UK;Neuromuscular and Regenerative Medicine Unit, University of New South Wales, Sydney, NSW 2010, Australia;Oncology Research Unit, School of Medical Sciences, University of New South Wales, Sydney, NSW 2010, Australia;Signalling Programme, Babraham Institute, Cambridge CB223AT, UK;The Garvan Institute of Medical Research, St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Sydney, NSW 2010, Australia;

关键词: intravital imaging;    pancreatic cancer;    breast cancer;    actin;    small GTPase RhoA;    FLIM-FRET;    biosensors;    immunology;    development;    cell biology;   

DOI  :  10.1016/j.celrep.2017.09.022

来源: DOAJ

【 摘 要 】

The small GTPase RhoA is involved in a variety of fundamental processes in normal tissue. Spatiotemporal control of RhoA is thought to govern mechanosensing, growth, and motility of cells, while its deregulation is associated with disease development. Here, we describe the generation of a RhoA-fluorescence resonance energy transfer (FRET) biosensor mouse and its utility for monitoring real-time activity of RhoA in a variety of native tissues in vivo. We assess changes in RhoA activity during mechanosensing of osteocytes within the bone and during neutrophil migration. We also demonstrate spatiotemporal order of RhoA activity within crypt cells of the small intestine and during different stages of mammary gestation. Subsequently, we reveal co-option of RhoA activity in both invasive breast and pancreatic cancers, and we assess drug targeting in these disease settings, illustrating the potential for utilizing this mouse to study RhoA activity in vivo in real time.

【 授权许可】

Unknown