Julie's Life

And so it begins...

Julie Earwood
Pathobiology and Molecular Medicine Summer Lab Rotation Abstract
Lab: Dr. Joan Cook-Mills
Lymphocytes migrate through endothelial tissues using pathways mediated by receptors, one of which is the binding of VCAM-1 to α4-integrin of the lymphocyte. The VCAM-1 then stimulates the calcium-and Rac1-dependent activation of NADPH oxidase. The NADPH oxidase produces superoxide that dismutates to H2O2, generating ROS. ROS activates MMPs, which degrade extracellular matrix and cell-cell junctions allowing cell retraction and the migration of lymphocytes.
We have observed the function of NADPH oxidase in vivo by using CYBB mice (gp91phox -/-) and wild type mice. Gp91phox is the catalytic subunit of NADPH oxidase. VCAM-1 dependent infiltration of eosinophils into the lung was induced by OVA intranasally. The mice were then sacrificed, the lungs were lavaged, and then the lungs were frozen for tissue analysis of leukocyte subsets. The tissue samples will be stained for major basic protein-1 (MBP-1) for eosinophil detection and stains for B and T cell surface markers. The tissue samples will then be counted to determine the number of lymphocytes within the tissue and blood vessels. Since the samples were lavaged, few leukocytes will be found in the alveolar cavities.
The importance of this experiment is to show that without NADPH oxidase activity, eosinophils cannot migrate across endothelial cells because VCAM-1 dependent ROS cannot be synthesized. Since ROS cannot be synthesized, MMPs would not be activated and there would be reduced retraction of the cell junctions. This could reduce inflammation caused by eosinophils in allergen-induced asthma.

So for you who are not trained in the biological sciences, this is the summary of what I will be doing this summer in my lab. I wrote it today as my first assignment in the lab. I basically will be counting cells in lung tissue slides on the microscope. I will also be helping the lab do bleeds for mice and at the end of the summer I will learn how to do western blots, which I don't understand enough right now to explain. It is very exciting and I can't wait to actually start it all.

I hope everyone has a wonderful weekend and a Happy July 4th!
Don't set your house aflame,
Juls

posted by Julie @ 6:08 PM