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  • Bigger Picture!
  • Animals with Whiskers – 2 types
  • What did I actually do? (follicle injections)
  • Now that we have the labeled tissue, used Confocal microscopy to get…

Transcript

  • 1. MAPPING OF WHISKER SENSORY INPUT IN THE TRIGEMINAL GANGLION OF THE MOUSE
    Stephanie Redmond
    David W. Matthews
    Dr. David Kleinfeld
    University of California, San Diego
  • 2. Mapping the Trigeminal Ganglion
    Where is the trigeminal ganglion (TG)?
    What makes up the TG?
    Whisker follicle injections
    Image analysis
    TG patterning
    Going forward
    Adapted from Kleinfeld, et al. 2008
  • 3. Where is the TG?
    Adapted from Kleinfeld, et al. 2006
  • 4. What makes up the TG?
    Many different cell types
    Whisker sensory neurons
    Temperature-sensing neurons
    Motor (movement) neurons
    Functions
    Correlations to vision
    Hyperacuity
    Trigeminal neuralgia
    Blood vessels rub against TG cells
    M
    R
    C
    L
    rostral(R), caudal (C),
    lateral (L), medial (M)
  • 5. Why should spatial organization exist?
    TG Studies:
    Anatomy
    Gene Expression
    Adapted from Waite 1992
    Adapted from Hodge 2007
  • 6. Whisker Follicle Injections
    Fluorescent retrograde neuron labeling
    Alexa Fluor® Cholera Toxin Subunit B
    Wild type mice, age ~P16
    TG histologicalslice
    50 to 75 micrometer slices
    488 nm
    555 nm
    647 nm
  • 7. Image Analysis
    • Confocal Microscopy
    • 8. Image through sample (~50-100 um)
    • 9. Develop image stacks
    Screen with pinhole
    Focal Point
    "Confocal Microscopy", D Semwogerere & ER Weeks, published in the Encyclopedia of Biomaterials and Biomedical Engineering, Taylor & Francis (2005).
  • 10. M
    R
    C
    L
    Sample E
    A (488) B (555) C (647)
  • 11. Image Analysis
    • Spatial Organization
    • 12. Measure cell height, width, length
    • 13. Calculate and compare average:
    • 14. Position, volume
    M
    R
    C
    L
    A (488) B (555) C (647)
  • 15. TG Patterning
  • 16. Going Forward
    • Process image data
    • 17. Investigate other patterns
    • 18. Average X-coordinate
    • 19. Average Y-coordinate
    • 20. Average volume
    • 21. Finish thesis paper
    Graduate
  • 22. Acknowledgments
    David W. Matthews
    Dr. David Kleinfeld
    The Kleinfeld Lab
    Physics REU Program
    UCSD
    MPS Faculty
    Professor Shilepsky
    Professor Heinekamp
    MPS Majors
    Wells College