Physical education (PE) lessons offer an excellent opportunity to encourage participation and learning for students with and without disabilities. However, there are still barriers that prevent educative inclusion (EI) from being achieved, with negative attitudes being one of the major issues. This study aimed to explore students without disabilities’ attitudes toward their peers with disabilities in the second stage of Primary Education, examining possible differences according to sex (male or female) and school location (urban or rural). The Scale of Attitudes toward Students with Disabilities in Physical Education–Primary Education (SASDPE-PE), a four-item instrument with a five-point Likert scale ranging from 1 (strongly disagree) to 5 (strongly agree), was administered to 545 Spanish students aged 9 to 12 years old. The results revealed that girls and students from rural schools showed more positive attitudes toward their peers with disabilities during the PE lessons; differences were significant in both cases. Thus, according to our findings, the SASDPE-PE is a practical tool to assess attitudes, even after an attitude-change programme. Furthermore, PE attitude-change programmes should be implemented, especially considering male students and those enrolled in urban schools.