Natural ingredients have been employed to develop food products.Hydrocolloids from butternut squash seeds (HBSSs) were extracted with water at pH 3, 7, and 10 and characterized bromatologically and rheologically; then these HBSSs were used to stabilize the dressing-type emulsion by evaluating its physicochemical, rheological, and microstructural properties.Hydrocolloids presented higher protein (from 20.43 to 39.39%) and carbohydrate (from 50.05 to 52.68%) content and rheological properties with a predominant elastic modulus. HBSSs extracted at pH 10 were used for the development of the dressing-type emulsion. The samples were stable during the storage period (15 days), with a good microstructural organization showing non-Newtonian fluid properties with shear-thinning behavior when the pseudoplasticity and the oil droplet size decreased with the addition of HBSS.Hydrocolloid constituents were detected surrounding the droplets of the emulsions, intensifying the effects of inner droplet interaction due to depletion events and a strong influence on the structure and physical stability. The hydrocolloids used to stabilize the dressing-type emulsions are additively promising in microstructured food design.