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The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells

The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells

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The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells

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  1. The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells By Amir Abyaneh, and Dr. Terrie Rife

  2. Nitric Oxide Synthase I • Nitric Oxide Synthase I (NOS I) is an enzyme that catalyzes the reaction of a free radical, NO•. • Learning and Memory • Neurodegenerative Diseases/Problems • Why do we study it?

  3. Nitric Oxide Synthase I

  4. Human NOS I Structure 5’2 exon 2 exon 3- 29 5’4 5’3 5’1 { >20 kB } Transcription Translation Note - Figure is not drawn to scale

  5. What We Know! Nerve Growth Factor NEURON p75 Trk A NOS 1 ↑

  6. Brain Derived Neurotrophic Factor

  7. p75 RA Trk B RA RA SH-SY5Y Cells RA RA Cell Lines • human neuroblastoma cells SH-SY5Y • A model for neuronal differentiation • Responds to BDNF after being treated with Retinoic Acid 5 days

  8. Untreated Establishing a Protocol RA w/ 10%FBS for 5 Days BDNF for No Serum for 6 days

  9. Methods • Pretreatment with RA and 10% FBS for 5 Days • Treated with BDNF for 2, 4, 6, and 8 days, with a No Treatment control • RNA Harvesting • Reverse Transcription • PCR Analysis

  10. Primers – PCR analysis RA 2 4 8 No DNA NT GADPH 380bp nNOS 433bp

  11. 5’2 exon 2 exon 3- 29 5’4 5’3 5’1 { >20 kB } Transcription Translation FUTURE • PCR analysis of transcripts: • 5’1 • 5’2 • 5’3 • 5’4 Note - Figure is not drawn to scale

  12. Thanks