Faculty
Ajith Karunarathne, PhD
Assistant Professor
Email: ajith.karunarathne@utoledo.edu
Office: BO 2098 A/B
Phone: (419) 530-7880
Fax: (419) 530-4033
Professional Background:
B.S.: University of Sri Jayewardenepura, Sri Lanka
PhD.: Michigan State University, East Lansing, Michigan, USA
Postdoctoral: Washington University School of Medicine, St. Louis, Missouri, USA
Research
Group Web Page
Publications
News
A new JBC paper from our group is just coming out. Congratulations authors!
Senarath, K., Payton, J. L., Kankanamge, D., Siripurapu, P., Tennakoon, M., and Karunarathne, A. (2018) Gγ identity dictates efficacy of Gβγ signaling and macrophage migration. Journal
of Biological Chemistry (in press)
THREE RECENT PUBLICATIONS FROM US JUST CAME OUT!
1. Siripurapu, P., Kankanamge, D., Ratnayake, K., Senarath, K., and Karunarathne, A.
(2017) Two independent but synchronized Gbetagamma subunit-controlled pathways are essential
for trailing-edge retraction during macrophage migration. J Biol Chem 292(42):17482-17495.
2. Ratnayake, K., Kankanamge, D., Senarath, K., Siripurapu, P., Weis, N., Tennakoon,
M., Payton, J. L., and Karunarathne, A. (2017) Measurement of GPCR-G protein activity in living cells. Methods Cell Biol 142, 1-25
3. Gupta, R. K., Swain, S., Kankanamge, D., Priyanka, P. D., Singh, R., Mitra, K., Karunarathne,
A., and Giri, L. (2017) Comparison of Calcium Dynamics and Specific Features for G Protein-Coupled Receptor-Targeting
Drugs Using Live Cell Imaging and Automated Analysis. SLAS Discov, 22(7):848-858
Our findings on itch sensation are now published in the journal SCIENCE SIGNALING. Click here to read it. Click here to read the UT news article on this research.
Our review article Titled " Subcellular optogenetics: controlling signaling and single cell behavior" is published
in JOURNAL OF CELL SCIENCES. Click here to read the article.
Our findings are published in the journal NEURON on cross-talks between Serotonin (5HT)- gastrin-releasing peptide (GRP) receptors
and subsequent effects on itch sensation. This new is on NBC news national coverage.
Click here watch NBC Brian Williams talking about this work. 
Click here to read this article.
Exciting undergraduate and graduate research opportunities!
We offer a science outreach program for high school students to conduct research in
life sciences!
Please contact Dr. Ajith Karunarathne for more information.
Research Interests:
Bioanalytical, Optical chemistry and Optogenetics, Molecular Imaging, and Signal Transduction
1. GPCR and G protein signaling in immune and cancer cell migration.
2. Engineering optogenetic approaches to control subcellular signaling.
3. Engineering of biosensors and assay to quantify dynamic behaviors of signaling
molecules.
4. Experiment-guided mathematical-computations modeling of cellular signaling networks.
5. Functional characterization opsin for their ability to control mammalian cell signaling.
6. Optical isomerization of push-pull molecules and the applicability in biomedical
research.
Research Synopsis:
Quantitative visualization of signaling in living cells surpasses cell disruptive
approaches by providing spatiotemporal information about molecular entities that govern
complex cellular processes. However, there is a lack of approaches that govern signaling
in a cell with precise spatial and temporal control to complement the existing assortment
of live cell imaging methodologies.
To bridge the above gap, research in our group interfaces chemistry and biology with
an emphasis on understanding signaling network dynamics in living cells. In order
to do this, we employ optical approaches not only to visualize but also to control
and interrogate signaling in single cells. We currently dissect several pathologically
important cell behaviors such as cell migration in cancer and neuronal damage repair.
To facilitate this process, we generate tools of two kinds using a combination of
chemical, genetic engineering, and molecular biology approaches. First, we design
optical triggers to govern entire or individual components of signaling pathways in
sub-cellular regions of single cells using specific wavelengths of light. Second,
we develop efficient sensors to probe the dynamics of signaling molecules through
fluorescence localization, bioluminescence or Förster resonance energy transfer (BRET
or FRET) techniques.
Using optical tools, we interrogate cell behaviors by activating, inhibiting or modulating
single, or multiple signaling modalities and simultaneously quantify the dynamics
of the associated cellular and molecular responses. To perform these experiments,
we employ single & multi-photon optogenetics and imaging approaches in combination
with two and three-dimensional microfluidic technologies and classical analytical
instrumentation. Additionally, we make use of a multiphoton confocal approach to perform
intravital optogenetics and imaging to control and visualize single-cell behaviors
in-vivo. These approaches will help us identify and understand molecules, mechanisms and their
dynamics in signaling networks that control cellular processes such as cancer metastasis,
demyelination in the nervous system and modulation of itch sensation. Our approaches
can be used to optically control a variety of cellular signaling and behavioral processes
in a whole animal as therapeutic strategies.
Optochemistry and G protein-coupled opsin activation to control cellular signaling:
A. 11-cis-retinal has lambda-max around 380 nm. However, the lambda-max of the 11-cis
retinal bound opsin varies from 350 nm (UV) to 640 nm (far red). During optical activation
of opsin, a photon with appropriate energy interacts with the covalently bound chromophore
11-cis retinal inducing its photo-isomerization to all-trans retinal. Conformational changes in opsin, due to this photo-isomerization, leads to
opsin activation.
B. GDP bound G protein a subunit has a higher affinity with the bg heterodimer. This
interaction leads to the formation of alpha-betagamma heterotrimer. Opsin interacts
with these heterotrimeric G proteins through its internal loops and the c-terminus
at the cytosolic surface. Activated opsin facilitates the exchange of GDP to GTP at
the G protein a subunit resulting in heterotrimer dissociation and formation of active
G protein subunits; alphaGTP and free betagamma. These activated G proteins are responsible
for signaling effectors activation and subsequent signal transduction.
There are a limited number of heterotrimer G proteins controls the majority
of cellular signaling and physiological processes. Our group focuses on employing
this G protein signaling conservation to achieve optical control of cellular and physiological
processes through the recruitment native and engineered opsins.
Optical control and visualization of subcellular signaling
HeLa cell in the movie expresses G protein-coupled vision receptors (Opsins) from
the retina of the eye. The cell also expresses mCherry tagged gamma-9 subunit. On
GPCR activation, G protein beta-gamma subunits translocate to the internal membranes.
In this experiment, opsins in the left side of the cell are exposed to a confined
blue light pulse (red box). Please note the disappearance of mCherry-gamma-9 from
the exposed plasma membrane region immediately after the pulse and appearance in the
adjacent ER. Opsin activation is transient and therefore, mCherry-gamma-9 returned
to the plasma membrane towards the end of the movie. This shows that opsin can be
used to spatially and temporally control signaling in subcellular regions in a cell(Subcellular
optogenetics). The plot shows the dynamics of mCherry-gamma9 in adjacent internal
membranes. Karunarathne et al, 2013, PNAS – E1565-E1574
