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Making organelles visible - in planta and in societas

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This is the presentation I gave in Salzburg at the Annual Meeting of the Society for Experimental Biology, July 2012, for receiving the President's Medal for Education and Public Affairs.

http://www.sebiology.org/meetings/Salzburg2012/pres_meds.html

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Making organelles visible - in planta and in societas

  1. 1. Dr Anne OsterriederSEB Salzburg July 2012 @AnneOsterrieder #seb2012
  2. 2. Golgi apparatus http://multimedia.mcb.harvard.edu/media.htm
  3. 3. http://youtu.be/cnK7RT1q0bAhttp://youtu.be/cnK7RT1q0bA
  4. 4. Why study the Golgi apparatus?• Central role in secretory pathway• Production of cell wall material• Fundamental research - but understanding these processes necessary to understand and improve plant productivity
  5. 5. Fluorescent proteins© Rosario Beach Marine Laboratory http://www.tsienlab.ucsd.edu
  6. 6. Golgi bodies in tobacco leaf cell labelled with green fluorescent protein (GFP)
  7. 7. How are Golgi bodies formed?How do they maintain their structure? (...especially during movement!)
  8. 8. By Jack and Jasons Pancakes (www.jackandjasons.com)
  9. 9. Osterrieder 2012, Journal of Microscopy
  10. 10. Characterising golgins with big lasers at the Central Laser Facility, STFC, Harwell 10 µm
  11. 11. FRET (Fluorescence Resonance Energy Transfer) We use FRET to study interactions between GFP- and mRFP-labelled proteins. Energy transfer occurs if:- donor fluorophore emission spectrum overlaps significantly with acceptor absorption- Fluorophores are in close proximity (1-10 nm) GFP-donor (D) and mRFP-acceptor (A) fused to proteins of interest (A and B) Sparkes et al. 2010, J. Exp. Bot.
  12. 12. FLIM (Fluorescence Lifetime Imaging Microscopy)• Monitoring lifetime of GFP by measuring the nanosecond decay of the fluorophore• Lifetime is unaffected by probe expression or excitation intensity BUT GFP lifetime is decreased by FRET!
  13. 13. Fluorescent lifetime maps visualise interactions between trans-Golgi proteins and small regulatory proteins 10 µmAverage lifetime : 2.5 ns Osterrieder et al. Traffic 2009
  14. 14. Fluorescent lifetime maps visualise interactions between trans-Golgi proteins and small regulatory proteins 10 µm 10 µmAverage lifetime : 2.5 ns 2.4 ns Osterrieder et al. Traffic 2009
  15. 15. Fluorescent lifetime maps visualise interactions between trans-Golgi proteins and small regulatory proteins 10 µm 10 µm 10 µmAverage lifetime : 2.5 ns 2.4 ns 2.1 ns (p= 1.14 x 10−22) Osterrieder et al. Traffic 2009
  16. 16. Transient interactions? 10 µmAverage lifetimes• AtVPS52: 2.4 ns• AtVPS52 + AtRAb-H1b: 2.3 ns• AtVPS52 + AtRAb-H1c: 2.4 ns
  17. 17. Exploring the ER-Golgi interface• Testing if cis-Golgi golgins tether Golgi bodies to the ER• Express fluorescent full-length and truncated versions• Confocal microscopy and optical trapping with laser tweezers• “Grab a Golgi” – work by Imogen Sparkes and Chris Hawes
  18. 18. Why is this important?Understand molecular mechanisms of Golgi stack structure Understand Golgi populations in individual cells Put Golgi populations into functional context in cells Study effects on plant phenotypes
  19. 19. http://youtu.be/evW93DtSoZY
  20. 20. http://www.plantcellbiology.com
  21. 21. Social networking sites
  22. 22. Are you ready to step out?
  23. 23. Start small…• Start with achievable projects• Be realistic about your skills and your time• Tie in with existing structures – Departmental newsletter – STEM Ambassador scheme – Volunteer at Science Festivals – Help with school workshops – Write article for a blog, school magazine… – Give a talk in a school, public lecture… – Sign up on social networking sites• Be specific about your content and audience
  24. 24. …but think big!• Build up teams with like-minded people (peers or interdisciplinary groups) and organise bigger projects. – “Brookes Science Bazaar” – university science festival – “Science writes to Life”, Pegasus Youth Theatre – collaboration with Oxford Brookes Poetry Centre and Fiona Sampson (professional poet
  25. 25. Make it part of your every-day work• Read something interesting? Post it to social media.• Get students to write summaries or articles about their research projects, lectures…• Upload short video clips to YouTube (microscopy movies, field trips…)• Get school science clubs to help you with your “real” research…
  26. 26. JUST DO IT!
  27. 27. AcknowledgementsOxford Brookes University MusicChris Hawes David Mansfield (Professor Science)Imogen Sparkes Cyrus MowerPlant Cell Biology Group Lorenzo Frigerio Charlotte CarrollCentral Laser Facility, STFCStan Botchway FundingChris Stubbs Oxford Brookes UniversityMark Pollard BBSRCAndy Ward …and many, many more whoWageningen University contributed toTijs Ketelaar movies, volunteered for events, helped withNorbert de Ruijters organisation…. http://www;youtube.com/plantendomembrane

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