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

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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

By Amir Abyaneh, and

Dr. Terrie Rife

nitric oxide synthase i
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?
human nos i structure
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

what we know
What We Know!

Nerve Growth Factor

NEURON

p75

Trk A

NOS 1 ↑

cell lines

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
establishing a protocol

Untreated

Establishing a Protocol

RA w/ 10%FBS for 5 Days

BDNF for No Serum for 6 days

methods
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
slide10

Primers – PCR analysis

RA

2

4

8

No DNA

NT

GADPH

380bp

nNOS

433bp

future

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

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