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The development
of cell-based biosensors in our laboratory is directed to detecting the
effects of several chemical welfare stimulants on the central nervous
system. The sensing systems developed are based on pheochromocytoma
cells, PC12, that have properties associated with immature neural crest
cells that are destined to evolve into adrenal gland chromaffin cells
or sympathetic neurons. These neuron-like cells are capable of
generating processes or extending projections (axons and dendrites)
from their cell body (soma) upon attachments to adhesion substrates
(extra cellular matrix proteins). The cessation of proliferation and
undergoing of differentiation upon attachment to the adhesion substrate
is insignificant. However, in the presence of specific trophic
substances, or hormones (i.e. nerve growth factor, NGF), the cells
undergo differentiation.
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Currently, we
use an ECIS 8 Well Electrode Array system for measuring the change in
electrical impedance of a small electrode to AC current upon seeding
the cells into individual wells. The cell culture chambers sit on an
array of gold film electrodes (200 m diameter) that connect to the
interrogation unit (instrument applying the current) to each of the 8
wells. The current across bare electrodes encounter low resistance,
however, upon the addition of cells, the cells attach and spread upon
the gold electrode surface and the current flow is now impeded or
restricted and the current must exist within the spaces under and
between the cells.
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There are many
compounds of interest to biological chemical welfare research. We are
particularly interested in those molecules that specifically bind to
cell surface receptors and affect the functionality of the CNS when the
receptor receives these ligands, such as sarin, mustard gas (nerve
gases) etc. As a result of binding, changes in a signal transduction
pathway are observed due to the enhancement or reduction of the
cellular response. Directly, with the use of a time course monitoring
of cellular response, we can avoid many potential false positives that
occurs with the assays that are concerned with binding. Cell-base
sensing provides more reliable information regarding the efficacy of
compounds and how the cells are affected over time.
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