While there is an abundance of research pertaining to the development of anxiety disorders, there is still a dearth of knowledge regarding the development of anxiety in the general population. The objective of this study was to longitudinally explore the development of mathematics anxiety among normative middle-school students, and to identify the moderating role of gender, school transition and scholastic achievements on these trajectories. Subjects included 413 sixth grade students (53.3% females, mean age 11.27 ± 0.38 years). Participants were evaluated for their level of anxiety in the context of mathematics, using the Value of Education scale, in four time-points with a 6-month gap between each time-point. Data regarding subjects' grades and school transition were also collected. A growth curve analysis using hierarchical linear modelling revealed that girls, students who transitioned between schools and high achievers (each independently) reported a significant increase in mathematics related anxiety towards the end of sixth grade, which later decreased during seventh grade. The findings of this prospective study on factors affecting the development of mathematics anxiety among normative adolescents may be important in planning focused primary prevention school-based strategies.