Results!   Leave a comment

Recently, I put up a 4-color picture, showing the nucleus (blue), GFP (green), somatostatin receptor-3 (red) and microtubules, especially the cilium (white).

Recently, I performed the same stain on a monolayer that was subjected to orbital shaking for 48 hours- the shaking induces fluid flow over the top of the cells. Here’s the two images, side-by-side (flow on the left, no-flow on the right):

Again, these images are “preliminary data”. I can’t draw definitive conclusions (yet). Here’s what is apparent:

1) In ciliated cells (cells with the strong white feature), SSTR3 lies in the nucleus when no flow is present, and in the cytosol when there is flow present.
2) the pattern of SSTR3 depends on whether a cilium is present in no-flow conditions- the no-flow unciliated cells look a lot like the flow cells, and there does not appear to be a difference between ciliated and non-ciliated cells in the presence of flow
3) The length of the cilium is shorter when there is flow present- that may not be glaringly obvious, but a careful measurement shows this.
4) When SSTR3 is detected at the nucleus, it has been cleaved from the GFP molecule. When SSTR3 is in the cytosol, it is co-located with GFP. This is shown by comparing where the green and red colors are within a cell. If you see yellow, that’s because the red and green colors are at the same location.

Notice, I have no idea which is ‘normal’. And I don’t know *why* SSTR3 would go to the nucleus, but I have seen the same effect from staining for Kif3a, a motor protein that is specific to the cilium.

This is why the results are preliminary- it’s a pure observation, I haven’t done any control experiments (to make sure the antibodies are actually binding to the correct protein, for example), etc. But, it’s good enough for a grant proposal! It demonstrates that I can do *something* and get *repeatable* results. I’m submitting a grant proposal next week, and these images (or something similar) will be in the proposal as, you guessed it, “preliminary data”.

On another front, Joey gave a talk at the regional meeting of the American Physics Society (did the name change become official yet..?) last weekend, and is presenting a poster at the regional meeting of the American Physiological Society tomorrow (Friday). Both of these are regarding his summer research project.

And I was just named to the Search Committee for the College of Science and Health Professions to locate a new Dean. I’m definitely excited about this opportunity, for a lot of reasons.

I’m hoping I get a chance to do some color interferometry today- two of my fluorescence filters should let me take some interesting photos of a fluid meniscus. As usual, I’ll post anything spiffy.


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