1 / 14

James Madison University Biology Department (Biosymposium)

James Madison University Biology Department (Biosymposium). Transcription Factors involved in the Upregulation of NOS I transcription by Phorbal Myristic Acid. Presented By Linda A. Yu. Date: April 21, 2001 Prepared for: Dr. Terrie Rife. What do all of these conditions have in common?.

donat
Download Presentation

James Madison University Biology Department (Biosymposium)

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. James Madison University Biology Department (Biosymposium) Transcription Factors involved in the Upregulation of NOS I transcription by Phorbal Myristic Acid Presented By Linda A. Yu Date: April 21, 2001 Prepared for: Dr. Terrie Rife

  2. What do all of these conditions have in common? • Stroke • Spinal Cord Injury • Alzheimer’s Disease • Parkinson’s Disease • Huntington’s Disease • ALS (Lou Gehrig’s Disease) • Mad Cow Disease

  3. Overview • Introduction of NO and NOS • Importance of NOS I • Relevance of PMA (Protein kinase C • pathway) • Procedures • Results and Conclusions • Improvement for the future

  4. Chemical Reaction of NO

  5. What does NO do? • Nitric Oxide • Intracellularmessenger • - Dichotomous effect • -Small amount: beneficial • -long-term memory • -synapse formation • -neuronal plasticity • -prevents cell death • - Large quantities: detrimental • -induces cell death

  6. What upregulates NOS I in neurodegenerative conditions?

  7. NGF upregulates NOS I and c-Jun C-Jun

  8. Protein Kinase C pathway • Protein Kinase C: activated by a chemical phorbal ester: phorbol myristic acid (PMA) • Serine/threonine kinase • Maximally activated in the presence of DAG (diacylglycerol) and calcium ions: mitogen activated protein (MAP) kinases, pkA and pkC. • Involved in the signal transduction pathways initiated by hormones, growth factors and neurotransmitters. (NGF pathway). • NGF pathway: signal cascades involved in upregulation of NOS I. • Transcription factors whose expression upregulated: • -two leucine zipper proteins: c-fos and c-jun; • AP1, AP2, and NFk-B.

  9. Procedures • Pheochromcytoma cells - PC12 cells derived from the adrenal gland of rats were grown in a monolayer and grown on a plate until 70-80% confluency. • Treatments: - DMSO - No treatment -PMA in DMSO (6 hours) -PMA in DMSO (8 hours) • Assay for Changes in NOS I Expression by RT-PCR

  10. Procedures (cont’d) • RNA isolation using Trizol Method. • RNA concentration measured : spectrophotometer. • RT-PCR (Reverse transcriptase PCR): to detect changes in gene expression. • PCRed cDNA in an 2% agarose gel: • Beta-actin:control • NOS1

  11. For the Future…. • If NOS I is upregulated following PMA treatment • Test to find the specific NOS I promoter that is upregulated by PMA • Isolate some of the transcription factors that are upregulated in the pkC pathway. • If NOS I is not upregulated following PMA treatment • Try to find other activation sites in the NGF pathway to see which transcription factors are upregulated when NOS I is upregulated. • Do further experiments on eNOS and iNOS.

  12. THE END

More Related