the effects of bdnf on nitric oxide synthase i using human sh-sy5y cells
DESCRIPTION
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 - PowerPoint PPT PresentationTRANSCRIPT
![Page 1: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/1.jpg)
The Effects of BDNF on Nitric Oxide Synthase I
Using Human SH-SY5Y Cells
By Amir Abyaneh, and Dr. Terrie Rife
![Page 2: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/2.jpg)
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?
![Page 3: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/3.jpg)
Nitric Oxide Synthase I
![Page 4: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/4.jpg)
Human NOS I Structure
Note - Figure is not drawn to scale
Transcription
Translation
5’4 5’3 exon 3- 29
{ >20 kB }
exon 25’15’2
![Page 5: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/5.jpg)
What We Know!Nerve Growth Factor
Trk A
p75
NOS 1 ↑
NEURON
![Page 6: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/6.jpg)
Brain Derived Neurotrophic Factor
Neurotrophin ReceptorNGF – Nerve Growth Factor
P75 and TrkA
BDNF - Brain Derived Neurotrophic Factor
P75 and TrkB
Neurotrophin 3 P75 and Trk C
Neurotrophin 4/5 P75 and TrkB
![Page 7: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/7.jpg)
Cell Lines human neuroblastoma cells SH-SY5Y A model for neuronal differentiation Responds to BDNF after being treated with Retinoic
Acid 5 days
Trk B
SH-SY5Y Cells
p75 RA
RA
RA
RA
RA
![Page 8: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/8.jpg)
Establishing a Protocol
UntreatedRA w/ 10%FBS for 5 Days
BDNF for No Serum for 6 days
![Page 9: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/9.jpg)
MethodsPretreatment with RA and 10% FBS for 5 DaysTreated with BDNF for 2, 4, 6, and 8 days, with a No Treatment controlRNA HarvestingReverse TranscriptionPCR Analysis
![Page 10: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/10.jpg)
Primers – PCR analysis
433bp
380bp
No DNA
NT 2 RA 4 8
GADPH
nNOS
![Page 11: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/11.jpg)
FUTURE PCR analysis of transcripts: 5’1 5’2 5’3 5’4
Note - Figure is not drawn to scale
Transcription
Translation
5’4 5’3 exon 3- 29{ >20 kB }
exon 25’15’2
![Page 12: The Effects of BDNF on Nitric Oxide Synthase I Using Human SH-SY5Y Cells](https://reader036.vdocument.in/reader036/viewer/2022062323/56815923550346895dc64d24/html5/thumbnails/12.jpg)
Thanks