Improving In Vivo Adenoviral Transduction for Detailed Studies of GLUT4 and AS160 Studies in Adipocytes

Obesity and type 2 diabetes are increasing at alarming rates worldwide, presenting significant public health challenges. A key player in these conditions is GLUT4, a protein that helps transports glucose into cells in response to insulin. Proper function of GLUT4 is crucial for maintaining healthy blood sugar levels and overall metabolic health.
To address this, in this thesis I have explored the molecular mechanisms influencing insulin action with a focus on GLUT4 trafficking in adipocytes. Given the limitations of traditional in vitro models, we optimized an in vivo adenoviral transduction method to express HA-GLUT4-GFP in perigonadal white adipose tissue. This approach allowed us to observe GLUT4 dynamics in freshly isolated adipocytes under basal and insulin-stimulated conditions using TIRF microscopy.
Using this method, we examined the role of AS160, a protein crucial for controling GLUT4 distribution. Our observations showed that in wilde type mice, insulin prompted GLUT4 to move to the cell surface, facilitating glucose uptake. However, in AS160-deficient mice, GLUT4 was already present on the cell surface without insulin stimulation. Thus, this method serves as a useful tool to accurately explore GLUT4 behavior, which could generate valuable insights into the mechanisms underlying insulin action and glucose uptake. Findings using this method could lead to better therapeutic strategies for managing and treating metabolic diseases like diabetes and obesity.