Molecular and muscle strip mechanics in health and disease
Linda Kachmar
Project :
I study the role of inflammatory cells on airway smooth muscle mechanics in asthma. I isolate splenocytes from human spleen tissue to purify the naïve cells, so I can then differentiate them into Th2 cells. After dissecting airway smooth muscle tissue, I incubate muscle strips with the Th2 cells for 24h to test their effect on smooth muscle mechanics by doing the quick release assay and dose response curves to obtain the velocity of shortening, force and % of relaxation in response to isoproterenol. I also test the effect of inflammatory cytokines from cell culture supernatant on airway smooth muscle strips. Flow cytometry is performed in collaboration with the Fixman lab. I also quantify cytokine production by ELISA and protein expression by Western Blot analysis.
In parallel to this, I help students and other members of the lab perform biochemistry experiments such as actin tagging, protein phosphorylation, protein extraction, gel electrophoresis and Western blotting.
Figure 3: Phos-tag gel of total extract phosphorylation in human airways.
