• Share
  • Email
  • Embed
  • Like
  • Save
  • Private Content

Loading…

Flash Player 9 (or above) is needed to view presentations.
We have detected that you do not have it on your computer. To install it, go here.

Like this presentation? Why not share!

Non-Genomic Actions of Steroids in Ovine Endometrium

on

  • 563 views

 

Statistics

Views

Total Views
563
Views on SlideShare
563
Embed Views
0

Actions

Likes
0
Downloads
0
Comments
0

0 Embeds 0

No embeds

Accessibility

Categories

Upload Details

Uploaded via as Microsoft PowerPoint

Usage Rights

© All Rights Reserved

Report content

Flagged as inappropriate Flag as inappropriate
Flag as inappropriate

Select your reason for flagging this presentation as inappropriate.

Cancel
  • Full Name Full Name Comment goes here.
    Are you sure you want to
    Your message goes here
    Processing…
Post Comment
Edit your comment

    Non-Genomic Actions of Steroids in Ovine Endometrium Non-Genomic Actions of Steroids in Ovine Endometrium Presentation Transcript

    • Exploring the Role of nER in Non-Genomic Estrogen Regulation of the Ovine Endometrium
      Kyle Ireton
      Dr. Fredrick Stormshak
      http://jeremy29.smugmug.com/photos/317349190_8xiJ8-X3.jpg
    • Relevance
      Biological Relevance
      Steroids regulate reproductive organs/processes and pituitary gland
      Health Related Issues
      Steroids regulate onset and development of breast and uterine cancer, and promote cardiovascular health
      http://physicians.truthaboutdialysis.com/wp-content/uploads/2008/02/doctor-with-patient.jpg
    • Two Regulatory Mechanisms of Steroids:Genomic (Slow) Vs. Non-Genomic (Fast)
    • Genomic actions of steroids:
      “Slow”
      http://scienceblogs.com/clock/upload/2006/11/a2%20steroid-receptor.jpg
    • Non-genomic actions of steroids:
      “Fast”
      http://www.scq.ubc.ca/wp-content/uploads/2006/07/transduction.gif
    • Possible Non-Genomic Actions of Estrogen
      Biological Basis for Investigation
      Estradiol (E2) is essential for rapid development of the endometrial lining, for reception of a fertilized ovum (egg)
      In Vitro Basis for Investigation
      Specific plasma membrane binding site for E2 in endometrium first observed by Pietras and Szego (1977)
      Translocation of up to 3% nuclear estrogen receptor (nER) protein from nucleus to plasma membrane demonstrated by Razandiet al. (1999) in transfected CHO cells
    • Research Focus
      Does a quantifiable correlation exist between nER and the membrane binding protein for E2, in a live domestic animal model?
      http://www.ks.uiuc.edu/Research/pro_DNA/ster_horm_rec/dbd/er-ere-system-big.gif
    • Working Hypothesis
      A strong, quantifiable correlation exists between nER and the membrane binding protein for E2, in the ovine endometrium.
      http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/M/MembraneProteins.gif
    • Methods
      Two groups ovariectomized ewes
      E2upregulatesnER production
      P4 suppresses nERproduction
      Koligian and Stormshak (1977)
      http://www.salmonellablog.com/43_flock_of_sheep.jpg
    • Injection Schedule
      Alternating injections simulate natural estrous cycle of ewes
      Greater levels on nER predicted in Group 1
      Hence, greater binding activity predicted for Group 1 ewes
    • Methods
      Inter-caruncular tissue of endometrium collected
      Tissue processed and E2 nuclear and membrane binding activity counted
      P.L. Senger, Pathways to Pregnancy and Parturition, First Revised edition, 1999
    • Tissue Sample Analysis
      Membrane Tissue Assays:
      BCA protein assay quantifies membrane protein
      Radioreceptor assay (utilizing [3H]- estradiol- 17β) quantifies specific binding activity of E2, per mg Protein
      Nuclear Tissue Assays:
      DNA assay quantifies DNA in nucleus
      Radioreceptor assay quantifies nuclear binding of E2, per femtomole DNA
    • Results:Nuclear Binding Activity of E2
      In units of fmol E2 bound/μg DNA
      P-value <.07
    • Results: Membrane Binding Activity of E2
      In units of fmol E2 bound/ mg membrane protein
      P-value <.05
    • Conclusions and Future Investigations
      Conclusions:
      Results support hypothesis that nER shares quantifiable correlation to E2 membrane binding protein, in live domestic animal models
      Future Investigations:
      Investigate blocking action of P4 on E2 nuclear/membrane binding (this Fall)
      Further elucidate intracellular mechanisms of non-genomic activity (ERK 1/2, PI3K, PIP2 pathways)
    • Acknowledgments
      Dr. Stormshak, Professor Emeritus and HHMI Mentor
      Mary Meaker, Lab Technician
      Brian Kitamura, HHMI 2005 participant
      Kevin Ahern, HHMI program Coordinator
      HHMI and URISC programs