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Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

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Page 1: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Identifying Genes Underlying Spinal Motoneuron Specification

Karan NathaniSPUR 2011

Judith Eisen LabMentors: Liesl Van Ryswyk and Dr. Steve Seredick

Page 2: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Overview

• Objective• Background• Summer Research• Conclusion• Future Directions

Page 3: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Overview

• Objective• Background• Summer Research• Conclusion• Future Directions

Page 4: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Motoneuron Specification

Goal: Understanding how motoneurons are specified

Types of Neurons

Page 5: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Implications of This Study

• Treatment of several Motoneuron Diseases

Spinal Muscular Atrophy patient

Page 6: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Why Zebrafish?

•Vertebrates•Transparent•Genetic•Tools

Page 7: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Overview

• Objective• Background• Summer Research• Conclusion• Future Directions

Page 8: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Road to Olfactomedin 1a

• Microarray screen

• Signaling Molecule

• Axon Pathfinding

Page 9: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Olfactomedin 1a Background

Nakaya and Tomarev 2007

Page 10: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Overview

• Objective• Background• Summer Research• Conclusion• Future Directions

Page 11: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Hypothesis

• Olfactomedin 1a plays a role in specifying the formation of motoneurons.

Questions•Olfactomedin’s Role in Motoneuron Specification?•Olfactomedin’s Function?

Page 12: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Methods

• RNA in situ hybridization• MO Injections• PCR

Page 13: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Assessing Morpholinos•Titration•Concentrations: 50, 125, and 500 µM

50µM 125µM 500µM

Page 14: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Overview

• Objective• Background• Summer Research• Conclusion• Future Directions

Page 15: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Results

Uninjected Transgenic Fish MO injected transgenic fish

Page 16: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

ResultsConclusion: Olfactomedin 1a may have a role in axon pathfinding!

Uninjected transgenic fish MO injected transgenic fish

Page 17: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Overview

• Objective• Background• Summer Research• Conclusion• Future Directions

Page 18: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Future Directions•Determine RNA Expression Pattern

•Further Studies of Protein Functions

Page 19: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Acknowledgements

Eisen Lab• Judith Eisen• Liesl Van Ryswyk• Steve Seredick• Julia Ganz• Levi Simonson• Ellie Melancon• Dayna Lamb• Fish Facility

Spur Program•Peter O’Day•Lyndsey Goforth•Blakely Strand•Toby Fish•Interns

REU

Page 20: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Thank you!

Page 21: Identifying Genes Underlying Spinal Motoneuron Specification Karan Nathani SPUR 2011 Judith Eisen Lab Mentors: Liesl Van Ryswyk and Dr. Steve Seredick

Splice-blocking Morpholino