o at a base level it would be interesting to deterimine how epthelail barrier funciton is affect by forces in Xenopus.
Why aren't tight junciton transmitting forces during cytokinesis?
and respond to the forces generated by cytokinesis and that the actin scaffold protein Anillin is a regulator of epithelial mechanics.
flares of the small GTPase Rho are capable of rapidly repairing localized breaches in the epithelial barrier of X. laevis embryos. I introduced a technique (ZnUMBA) that allowed us to visualize transient leaks in the tight junction barrier for the first time, and correlated those events with changes in cell shape and localized loss of the tight junction proteins ZO-1 and Occludin. Then, I showed that flares of active Rho mediate repair of the barrier through localized contraction of the junction. In the field of tight junction signaling, this work raises many questions, including why use Rho for tight junction repair? How will our knowledge of Rho-mediated repair influence our treatment of diseases resulting from increased paracellular permeability? What are the GEFs, GAPs, and scaffolding proteins that regulate Rho flares? How does local loss of ZO-1 and Occludin, but not Claudin, cause barrier breaches? In this chapter I will speculate on each of these topics and propose what I see as the most exciting future directions to follow up on this work
brief review of what dissertationis about
including basic questions about brought up by the work
then cover those question in detail
Cell division stuff
1 A study of cytokineis in many different tisssues, across many different organisms. DIfferent types in highly proliferative epithelial vs non? Just looking at actin and myosin. In cancer?
2 how stiffness affects cell division (cancer)
3 more robust fucking up of vinculin and barrier function during cell division evernote. Use various mutants of Vinculin
4. Balance of tension in dividing cells?
5. Do neighboring cells assist in cell division in different ways compared to droshopila
6. More sensitive barrier function assay, association of flares around dividing cell
7. Differences in actin dynaimcs in epithelial dividing cells vs isolated
8. Why don't tight junctions feel the force?
Anillin and mechanics stuff
1. How does Anillin localization affect tumor progression? Extremely nuclear localization vs a NLS mutant. Tumor model. Tadpole?
2. Anillin's role in development. Neurulation, gastrulation.
3. Teasing apart junctional vs medial-apical contributions of Anillin.
n
5. In vitro expeirments with Anillin, bundling vs RhoA actiavtion effects. (Purify Anillin), super res, deep etch EM with Anillin OE KD
6. Direct test of Anillin's tention sensitivity (Fret purified Anillin)
7. Modeling the function of Anillin's contributions to organizing F-actin in epithelial cells.
8. Communication between Junciton F-actin and medial-apical.