Regulation of ASK3 inactivation under hyperosmotic stress

Abstract

Abstract Background: Hypertension today is common and a great risk factor for cardiovascular disease (CVD). WHO states that more people die annually from CVDs than from any other cause. This project focuses on the regulation of apoptosis signal regulating kinase 3 also known as ASK3, which has been observed to regulate blood pressure in the kidney. To clarify the regulatory mechanism of ASK3 activity is important for the development of new drugs against hypertension. It has been suggested that transient receptor potential cation channel, mucolipin (TRPML1), a non selective cation channel, might be the upstream regulator of ASK3. Objectives: To investigate whether TRPML1 is necessary for the regulation of ASK3 under hyperosmotic stress through different laboratory techniques. Methods: The research was conducted at The University of Tokyo, laboratory of cell signaling during September to November 2016. To answer the research question many different laboratory technics were used. Such as immunoblotting, immunostaining, immunoflourecense and cell culturing. Results: One of the five experiments supported the hypothesis that TRPML1 would be the upstream regulator of ASK3 under hyper osmotic stress. Another experiments gave negative results, thus one could not draw the conclusion that TRPML1 is a regulator of ASK3. A TRPML1 KO cell line was also established for further experiment in this field. Conclusion: Three months is not enough to draw any conclusions regarding the regulation of ASK3 but the results showed some promise. The experiments needs to be repeated and re-evaluated. The experiment that gave negative results could be due to my short experience in the lab, so if done