This chapter describes various approaches allowing the study of hyperosmolarity in the functions of 3T3-L1 adipocytes. Hyperosmolarity mimics insulin responses, such as glucose uptake and membrane ruffling, but also antagonizes these insulin effects, which can be evaluated in 3T3-L1 adipocytes. The molecular mechanisms of these effects can be also investigated by measuring the activation of different signaling pathways: (i) the phosphorylation of docking proteins on tyrosine and serine residues (serines 307 and 632), (ii) the phosphorylation of serine/threonine kinases, and (iii) the activation of phosphatidylinositol 3-kinase.