| Cell
shape is certainly the most distinguishing histodiagnostic feature of
differentiated cells and changes thereof are indicative of altered
genetic
programs as they occur physiologically during development and
specification but
also in pathological situations such as malignant tumour formation,
which is
coupled to dedifferentiation and migration. These processes necessiate
pronounced restructuring of cells which is determined by the
cytoskeleton. It
consists of three major filament networks that are composed of actin,
tubulin
and intermediate filament proteins, respectively. Thus, an intricate
relationship exists between regulated cytoplasmic cytoskeletal dynamics
and
cell migration/differentiation. Our own experiments in living cells
producing
fluorescent cytoskeletal filaments revealed coordinated and
interdependent
dynamic behaviour of all three filament systems albeit with different
kinetics and
distribution features and showed that signalling regulates their
restructuring.
Fluorescent fusion proteins will be used to label each of the three
major
cytoskeletal networks in gastric cancer cell lines, i.e. actin fusions
for
actin filaments, keratin 18 fusions for intermediate filaments, and
tubulin
fusions for microtubules. To facilitate detection in various
combinations in
the same cells, three different fluorescent proteins will be employed
that can
be individually detected. We will record the dynamic alterations of the
different cytoskeletons under conditions that will enhance or inhibit
motility.
The goal is to define quantitative measures to describe
multidimensional
cytoskeletal dynamics in response to different stimuli. |
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