I titled this post practice/plan because this week I’ve basically spent all my time working on the western blots (dry runs for my GR assay) and starting to hone in on on the context I will be looking at GR in. I developed the first batch of blots on Monday and Tuesday using a new chemiluminescence technique that was pretty neat but yielded a very weak signal for both gels.
I plan to run at least two more practice gels, which will hopefully yield a stronger signal next week.
In other news, I’m finally starting to put together a plan for the rest of my time at HPL! Right now, its basically centered around a modest research project to correlate GR sensitivity/expression level to obesity and insulin resistance in African American (AA) subgroups. If I can find such a relationship, it would lend itself to a greater understanding of how HPA (hypothalamic-adrenal-pituary) dysregulation is related to insulin resistance and obesity since insulin is an antagonist to cortisol and both insulin resistance and obesity affect AA at higher rates than Caucasians (CA). This is essentially based on literature that says HPA dysregulation can result from chronic stress and AA experience disproportionate levels of chronic stress and insulin resistance. I am trying to see whether differential GR sensitivity can help link the two. In other words, can the hypothalamuses’ inability to respond to negative feedback inhibition be linked to insulin resistance through GR sensitivity. While GR sensitivity can be influenced by several things besides competing ligands such as insulin, I believe quantifying GR in different AA subgroups offers an excellent opportunity to discover a real relationship. Like I said in the last post, the GR kits are supposedly on the way so as soon as they arrive and I can prove to Joel my western technique is up to bar, things should start to happen. (Even though I will be using ELISAs to look at GR, westerns are similar enough in concept that Joel is letting me prove myself with them using estrogen receptors). If everything goes my way, I will be using a cohort of 100 AA, who will hopefully represent several subgroups (obese, normal weight, insulin resistant, chronically stressed..etc.)