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Mapping of Vibrissa Sensory Input in the Trigeminal - MAPPING OF ...

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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 R M L C 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 histological slice – 50 to 75 micrometer slices 488 nm 555 nm 647 nm
  • 7. Image Analysis • Confocal Microscopy • Image through sample (~50-100 um) • Develop image stacks "Confocal Microscopy", D Semwogerere & ER Weeks, published in the Encyclopedia of Biomaterials and Biomedical Engineering, Taylor & Francis (2005). Screen with pinhole Focal Point
  • 8. Sample E A (488) B (555) C (647) R M L C
  • 9. Image Analysis • Spatial Organization • Measure cell height, width, length • Calculate and compare average: • Position, volume A (488) B (555) C (647) R M L C
  • 10. TG Patterning 0 200 400 600 800 1000 1200 A B C D E SquareMicrometers Whisker Average Cell Cross-sectional Area 1 2 3 4 5
  • 11. Going Forward • Process image data • Investigate other patterns • Average X-coordinate • Average Y-coordinate • Average volume • Finish thesis paper • Graduate
  • 12. Acknowledgments David W. Matthews Dr. David Kleinfeld The Kleinfeld Lab Physics REU Program UCSD MPS Faculty Professor Shilepsky Professor Heinekamp MPS Majors Wells College