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Page 57
allied
academies
Cell Science, Stem Cell Research &
Pharmacological Regenerative Medicine
November 29-30, 2017 | Atlanta, USA
Annual Congress on
Adv cel sci tissue cul 2017 | Volume 1 Issue 2
Our lab works to determine the mechanism whereby the
planar cell polarity (PCP) protein Vang-Like 2 (Vangl2)
regulates cell migration during embryogenesis. We focus
on the gastrula stage of zebrafish development as the cells
naturally undergo PCP-dependent migration. Loss of Vangl2
in trilobite mutant embryos results in a strong convergence
and extension phenotype characterized by shortened
and broadened body axes. Here, both ectodermal and
mesodermal cell populations fail to polarize. Previous data
established migrating vangl2 mutant cells lack directionality
and meander compared to wild type. We have also shown
vangl2 mutants have increased matrix metalloproteinase
activity and decreased fibronectin extracellular matrix
(ECM). We hypothesize defective cell-ECM interactions
underlie the vangl2 mutant phenotype. Using time-lapse
confocal imaging, we have now analyzed the membrane
protrusive activity of ectodermal cells fromwild-type and PCP
mutant embryos. Our current data suggest vangl2 mutant
ectodermal cells exhibit increased membrane protrusive
activity and have significantly fewer polarized protrusions.
Our data suggest filopodia are concentrated at the trailing
edge in wild-type cells, while vangl2 mutant cells have more
filopodia at the leading edge. We also found that vangl2
mutant ectodermal cells have reduced directness compared
to wild type. To determine the requirement for fibronectin
during protrusion formation, we used morpholinos to
knockdown fibronectin protein expression in wild-type
embryos. The data showed that fibronectin morpholino
injected cells exhibited increased formation of non-polarized
membrane protrusions similar to vangl2 mutant cells,
suggesting defective cell-ECM interactions contributing to
at least a portion of the mutant phenotype. Our preliminary
studies suggest decreased Vangl2 protein localization to
filopodia and larger membrane protrusions. Together, our
data suggest a model whereby Vangl2-dependent regulation
of cell-ECM interactions is required to suppress inappropriate
proper membrane protrusive activity.
e:
Vangl2 regulates membrane-protrusive activity in migrating gastrula cells
Anna Love and Jason Jessen
Middle Tennessee State University, USA