APR 30, 2020 8:00 AM PDT

Uncovering the membrane mechanism of cytokinesis using live cell imaging and subcellular optogenetic tools.

Sponsored by: Andor
C.E. Credits: P.A.C.E. CE Florida CE
Speakers
  • Product Specialist for Microscopy Cameras at Andor Technology.
    Biography
      My background in microscopy is built from my work and studies while at Queen's University Belfast. I always had a keen interest in bacteria and viruses as pathogens, as well as how we may exploit them for our benefit, so naturally specialised in microbiology. After my BSc (Hons) in 1999, I went on to do my PhD in microbiology within the School of Biology and Biochemistry at the Medical Biology Centre at Queens. My PhD was focused on polyphosphate metabolism of microorganisms- trying to get a better understanding of this molecule, how to quantify it, and the key roles this played within environmental and pathogenic bacteria. Roles include as a protective function in response to stress and in helping pathogens establish infections. An interesting application of this was in biological phosphate removal from wastewater for which we had a pilot scale study. This then led on to a Research Fellow position in the same research group of John Quinn and John McGrath in 2002. While in this position I worked further on connecting the biochemistry and genetics of polyphosphate metabolism. This included studies to characterise the bacterial transport systems involved and development of novel research tools, such as fluorescent assays to determine intracellular pH, and how to apply our luminescence and fluorescence imaging based methods to 96 well plate formats for faster screening and analysis. After Academia, I went on to work in the Medical Diagnostics and Pharmaceutical industries. I joined Andor Technology in 2012, initially working as a Technical Author for our camera and systems products. My current role is a Product Specialist for microscopy cameras. This role brings me in touch with the latest developments in camera technology, as well as a broad range of microscopy applications and techniques.
    • N. Gautam lab. - Department of Anesthesiology, Washington University School of Medicine in St. Louis, MO
      Biography
        Jean graduated as a Biotechnology Engineer from Instituto Politecnico Nacional in Mexico. He then received a PhD in Biochemistry from Universidad Nacional Autonoma de Mexico, where he worked with GPCRs and signal transduction. Currently, Jean is a Postdoctoral Research Associate at the Gautam lab in Washington University School of Medicine in St. Louis. He is using subcellular optogenetic tools to study processes like cell migration and cytokinesis.
      • Product Specialist - Life Sciences, Andor Technology
        Biography
          Claudia graduated in Applied Chemistry, has a MSc in Human Molecular Biology, a PhD in Cell Biology, and two postdocs in Cell Biology.
          With more than 15 years' experience in research, Claudia has worked in several areas like cell biology, HIV studies, oncobiology, endocrinology and neurosciences. In 2010 Claudia joined Universidade do Algarve as a facility manager and three years after Claudia become the director of Advanced Light Microscopy Facility.
          In May 2019, Claudia joined Andor Technologies as a Product Specialist in the Microscopy Systems Division.

        Abstract
        DATE:   April 30, 2020
        TIME:   8AM PT, 11AM ET, 4PM BST, 5PM CET
         
        Cytokinesis is the physical separation of two cells that occurs after the completion of mitosis. The mechanism underlying it is very complex. Until recently, the processes that lead to cleavage furrow formation and the extensive membrane remodelling that culminates in cytokinesis were unknown. This was due to the inexistence of methods to rapidly induce cytokinetic processes at will while simultaneously imaging membrane dynamics with high spatiotemporal precision.
         
        In this webinar, we will present how we used a combination of optical techniques such as subcellular optogenetics, FRAP, TIRF, confocal imaging (Dragonfly) and SRRF-stream imaging to uncover the membrane dynamics during the final steps of cytokinesis. By optically activating the small GTPase RhoA in the middle of a cell, we were able to induce cleavage furrow formation, and continued activation resulting in elongation of the furrow to form a structure resembling the intercellular bridge. Real-time confocal imaging showed that furrow formation is mediated by actomyosin contractility, plasma membrane flow to the furrow, localized decrease in membrane tension, and increased endocytosis in the furrow. Furthermore, FRAP experiments showed exocytosis in the intercellular bridge-like structure. TIRF combined with SRRF-stream imaging revealed a second exocytosis mechanism involving incorporation of membrane lipids from vesicles into the plasma membrane at the front edge of the nascent daughter cell. These results allowed us to propose a model in which spatially separated but coordinated plasma membrane depletion and addition mediate membrane remodelling during cytokinetic processes.
         
        In this webinar, the following questions will be addressed:
        1. What are the membrane remodelling events that occur during cleavage furrow formation?
        2. What is SRRF-Stream?
        3. How can I to use optogenetic tools to analyse biological events?
         
        Learning Objectives:
        • Understand the role of vesicular trafficking in the membrane remodelling process during cleavage furrow formation.
        • Discuss the advantages of using spinning disk confocal and optogenetic tools when researching dynamic events.
        • Identify different microscopy techniques such as TIRF and FRAP, SRRF-stream, and their applications.
         
        Webinars will be available for unlimited on-demand viewing after live event.
         
        LabRoots is approved as a provider of continuing education programs in the clinical laboratory sciences by the ASCLS P.A.C.E. ® Program. By attending this webinar, you can earn 1 Continuing Education credit once you have viewed the webinar in its entirety.
         
         

         

         


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