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Rick T. Mathias, Professor
Professor
Mathias with project assistant Abha Kochhar. Research in biophysics seeks physical insights into how cells and tissues
perform their marvelous repertory of functions, with the ultimate goal to better the
health of mankind. In our lab, research is directed toward understanding two health
problems: the formation of cataracts in the elderly and cardiac arrhythmias that can lead
to sudden death. Though these sound completely unrelated to one another or to biophysical
properties, our work suggests both are related to membrane transport proteins, membrane
voltage and ionic current flow from cell to cell. My early work was on the voltage
distribution and 3-dimensional current spread in a multi-cellular tissue. Because of my
undergraduate and graduate education in the physical sciences and mathematics, I found
this a fascinating problem that started me on a career of biomedical research. Maxwell's
classical equations relating charge and voltage provide part of the picture but ions move
about by diffusion and convection as well as conduction, so the equations of
thermodynamics, describing the coupling of these driving forces to ion fluxes, also apply.
What I found most striking, however, was the degree of control evolution has over the
pattern of flow. The geometry of the cells, the interconnection between cells and the
specific membrane transport proteins in local groups of cells provide enormous
flexibility. In the last 10 years, we have focused on the roles of specific membrane
proteins that generate and direct fluxes of ions, water or neutral solutes.
recent publications from the Lab...
George
J. Baldo
He is recording the frequency domain impedance of an intact
lens. Microelectrodes are used to inject random current into one cell and the record
voltage in other cells at various locations. A Fast Fourier Analyzer determines the
impedance in real time. This technology is used in connection with modern genetics that
produces mice lacking specific lens membrane proteins, "knock outs". The
impedance technology determines the effect of a "knock out" on the electrical
properties of the lens.
Junyuan Gao
He is using the whole cell patch clamp technique to determine membrane
transport properties of cells isolated from guinea pig hearts. A glass pipette is sealed
to the cell, then the underlying membrane ruptured to allow diffusion from pipette to
cell. This allows control of the chemical composition and the voltage within the cell. The
membranes of these cells contain a variety of transport proteins. There are ion channels,
that allow electrodiffusion of specific ions, ion pumps, that use the energy stored in ATP
to pump specific ions against their electrochemical gradient and exchange transporters
that use the energy in the electrochemical gradient for one ion to exchange transport a
second ion. The patch clamp is used to determine the role of each protein in the
electrical activity of the heart.
Xiurong Sun
She is using the whole cell patch clamp technique to study
the transport properties of isolated lens epithelial cells. The lens is composed of two
types of cells: fiber cells make up the mass and epithelial cells cap the front
hemisphere. We mechanically separate these two classes of cells and isolate the epithelial
cells for patch clamp analysis. We have been unable to isolate individual fiber cells so
we instead identify and clone specific fiber cell membrane proteins and study their
function in the oocyte expression system. For the epithelial cells. however, we can
directly measure the transport properties of membrane ion channels, pumps and exchange
transporters. These proteins help establish standing ionic currents and fluid movement
that circulate through the normal lens. We are interested in how these fluxes are
generated and their role in homeostasis.
Raj Varadaraj
He is measuring the rate of volume change of membrane vesicles when the
external osmolarity is abruptly changed. Images of the vesicle are saved using digital
video microscopy and later analyzed to determine the water permeability of the membrane.
Vesicles are made from cell membrane extracted from normal and "knock out" mouse
lenses. This allows us to determine which proteins are involved in membrane water
transport.
Sindhu Kumari
She is
carrying out biochemistry measurements of lens
connexins and aquaporins. Site directed mutagenesis, along with RT-PCR,
heterologous expression and immunohistochemistry are used to evaluate lens
connexins and the membrane proteins involved in membrane water
transport.
Francisco J Martinez-Wittinghan
He is
using the dual two
electric voltage clamp (Dual TEV) setup to study coupling between Xenopus
laevis oocytes expressing connexins. This technique allows to study
the pH regulation of the lens gap junction proteins connexin 46 and Connexin 50.
This technology is used in connection with impedance studies in the intact lens
to asses the coupling of the lens fibers in mice lacking specific lens membrane
proteins, "knock outs" and "knock ins".
Huan
Wang
Graduate Student
Wei Wang
Graduate Student
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