1 / 22

Presented by Juan Mo

Presented by Juan Mo. Study Questions. Why zebrafish transgenetic lines are very important for studying photoreceptor degeneration? The synapses in inner plexiform layer and in outer plexiform layer? ON bipolar cell & OFF bipolar cell?

ursala
Download Presentation

Presented by Juan Mo

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Presented by Juan Mo

  2. Study Questions • Why zebrafish transgenetic lines are very important for studying photoreceptor degeneration? • The synapses in inner plexiform layer and in outer plexiform layer? • ON bipolar cell & OFF bipolar cell? • What are the larval and adult phenotypes of pde6cw59 and Xops:mCFPq13 lines? • Light and dark preference test? • How does morpholino injection knock down gene expression?

  3. Zebrafish: outstanding model studying photoreceptor degenerations larvae adult • Contains red, green, blue and UV cone cells • Contains rods • Advantages: • Ex utero rapid development • Optical transparency • Permeability to drugs • Ease of making transgenetic animal • The acute visual abilities of larvae

  4. pde6cw59 transgenic line cone degeneration death of rod (larvae) rod persistence (juvenile and adult) Xops:mCFPq13 transgenic line rod degeneration loss of cone (mammanlian) cone unaffected (zebrafish)

  5. Hypothesis • Rod photoreceptors protects from cone degeneration-induced retinal remodeling and restore visual responses in zebrafish

  6. http://what-when-how.com/neuroscience/visual-system-sensory-system-part-2/http://what-when-how.com/neuroscience/visual-system-sensory-system-part-2/

  7. metabotropic glutamate receptor ionotropic glutamate receptor Na+ K+ http://what-when-how.com/neuroscience/visual-system-sensory-system-part-2/

  8. A B M ON ON OFF OFF ribbon http://webvision.med.utah.edu/book/part-ii-anatomy-and-physiology-of-the-retina/oute-plexiform/

  9. Confocal Microscopy

  10. Result 1: development of ON bipolar cell contacts with rod and cone photoreceptors nyx:mYFPq16 cones rods OPL ON bipolar cell ON bipolar cell Cone pedicle Rod spherule dendrites cones rods

  11. pde6cw59 transgenic cone degeneration Xops:mCFPq13 transgenic rod degeneration Result 2: cone degeneration in pde6cw59 mutant larvae leads to ON bipolar cell remodeling cones ON bipolar cell ON bipolar cell rods ON bipolar cell ON bipolar cell

  12. pde6cw59 transgenic cone degeneration Xops:mCFPq13 transgenic rod degeneration Result 2: cone degeneration in pde6cw59 mutant larvae leads to ON bipolar cell remodeling D6 ON bipolar axon terminals 6 DPF D8 8 DPF

  13. pde6cw59 transgenic cone degeneration OFF bipolar cell ON bipolar cell Xops:mCFPq13 transgenic rod degeneration horizontal cell ON bipolar cell Result 3: postsynaptic glutamate receptor expression is altered in pde6cw59 larvae D6 6 DPF D8 8 DPF

  14. Result 4: synaptic structure is maintained in Xops:mCFPq13 and pde6cw59 adults cones cones cones ON bipolar cell ON bipolar cell ON bipolar cell ON bipolar cell ON bipolar cell ON bipolar cell

  15. Result 4: synaptic structure is maintained in Xops:mCFPq13 and pde6cw59 adults photoreceptor terminals photoreceptor terminals photoreceptor terminals horizontal cell horizontal cell horizontal cell Cone pedicle rod spherule

  16. Result 5: visual responses are preserved in pde6cw59 mutant adults diffuse white light

  17. Result 5: visual responses are preserved in pde6cw59 mutant adults • Optokinetic response (OKR): http://www.youtube.com/watch?v=mfstlvsW1IY • WT and pde6cw59 adults display a robust OKR.

  18. Result 6: increased rod number protects the larval retina from cone degeneration- induced remodeling nuclear rod ON bipolar cell

  19. Morpholino: knock down gene translation

  20. Conclusion • Cone degeneration leads to ON bipolar cell remodeling in larvae. • Although the neural connectivity remains intact, the loss of the majority of the presynaptic input leads to significant remodeling observed in cone degeneration adult. • Visual responses are preserved in cone degeneration adult. • Increased rod number protects the larval retina from cone degeneration-induced remodeling. ? Rod photoreceptors protects from cone degeneration-induced retinal remodeling and restore visual responses in zebrafish

  21. Review Questions • Why zebrafish transgenetic lines are very important for studying photoreceptor degeneration? • The synapses in inner plexiform layer and in outer plexiform layer? • ON bipolar cell & OFF bipolar cell? • What are the larval and adult phenotypes of pde6cw59 and Xops:mCFPq13 lines? • Light and dark preference test? • How does morpholino injection knock down gene expression?

More Related