Primary Research Focus
Kidneys play a crucial role in maintenance of pH, water, and electrolyte balance in our body. The renal epithelial cells that perform the ion and water reabsorption are diverse in nature. Chandrasekar lab is interested in understanding the role of the actin cytoskeleton and associated myosin motor proteins in regulation of renal epithelial transport pathways and transport related kidney disease. We use a multifaceted approach involving conditional knockout mouse models, cell biological, biochemical methods including advanced microscopy techniques. The lab is also involved in development and application of novel tools and techniques to study membrane remodeling, organelle structure and function in vitro in cell culture and in vivo in mouse and human kidneys.
About the Chandrasekar Lab
Role of Myosin motor proteins in regulating cargo transport
Our previous work demonstrated that the loss of some proteins in mouse renal epithelium resulted in abnormal transport of thick ascending limb (TAL) cargo proteins, sodium, potassium, and a chloride cotransporter which led to progressive kidney disease. We utilize this mouse model and a new TAL-specific mouse model to further investigate the mechanisms by which these molecular motor proteins regulate cargo transport and influence ion reabsorption in the kidneys.
Novel immortalized thick ascending limb epithelial cell culture system to study cellular transport in vitro.
We generated an immortalized thick ascending limb (imTAL) cell line from the adult kidneys of mice that had been genetically modified with the immortomouse transgene. We perform biological and biochemical testing to assess the anterograde transport of cargo proteins and perturbations in the organelle transport pathways.
Conditional Myh9 and Myh10 inactivation in adult mouse renal epithelium results in progressive kidney disease
In 2020, our lab published a paper in the JCI Insight journal that highlights the critical role of actin-associated myosin motor nonmuscle myosin II (Myh9 and Myh10) in regulating specialized renal epithelial cargo transport machinery and kidney function. Our work established Myh9 and Myh10 playing a causal role in renal tubular disease, which is of significant interest since mutations in the MYH9 gene in humans result in kidney disease in at least one-third of patients.
3D reconstruction of organelles in the mouse kidney
We hypothesize that some proteins may regulate endoplasmic reticulum transport. To address this, our goal is to develop and standardize techniques and tools to visualize endoplasmic reticulum structure and dynamics in adult mouse renal epithelial cells using new 3D reconstruction technology.
Generation of Novel Biosensor to Measure Intracellular Tension
We have successfully generated a FRET-based biosensor of NMII that enables us to measure force generated along the actin filaments in live cells. We collaborate with Dr. Jing Liu, an expert biophysicist at Indiana University for this work. In addition to the biological questions, we are addressing using this biosensor; this novel tool can be of use to scientists from various fields such as cancer biology, bioengineering, material science etc.
Meet the Chandrasekar Lab Team
Karla Otterpohl, PhD
Karla is involved in developing and characterizing the various genetic knockout mouse models we use in the lab. She also performs advanced imaging techniques like TIRFM and live cell confocal imaging to interrogate the role of NMII isoforms in cellular transport pathways. Karla is an expert mouse geneticist with extensive training in genetic mouse models and molecular biology techniques. In the Chandrasekar Lab she has worked on expanding her skill set to advanced microscopy techniques and cytoskeletal research. She earned a PhD from the University of Nebraska Medical Center in 2015.
Brook W. Busselman
Brook joined the Chandrasekar lab in May 2019. His responsibility is to explore the molecular link between three human kidney disease-related genes: uromodulin (UMOD), Sodium potassium chloride cotransporter (NKCC2) and Nonmuscle Myosin IIA (MYH9). He works with conditional genetic knockout mouse models, novel mouse thick ascending limb cells generated in the lab to address the questions. Brook completed his undergraduate studies in medical biology at University of South Dakota. He has several years of work experience in pre-hospital emergency medicine and has gained other lab experience. He is currently pursuing a doctoral degree in biology and was accepted into the University of South Dakota’s Basic Biomedical Sciences Graduate Program.
Jenna joined the Chandrasekar lab in June 2021. She is using biochemical techniques to investigate the role of NMII isoforms in the kidney. She earned a bachelor’s degree in biology from the University of Wisconsin-La Crosse in 2021.