Like vertebrates, invertebrates such as Drosophila display complex integrated behaviors that rely on locomotion for their execution. The use of genetic tools combined with sophisticated behavioral analysis has permitted researchers to investigate the brain structures implicated in those complex behaviors, such as locomotor activity. The video-tracking paradigm has allowed the study of multiple parameters of locomotor activity and has revealed that Drosophila exhibits centrophobism, a behavior related to spatial orientation. A structure/function study has demonstrated that the mushroom bodies (MBs) are implicated in this behavior. In the continuity of these former studies, we have investigated the role of the cAMP transduction pathway known to be implicated in olfactory learning and memory within the MBs. Here, we report that disturbing this pathway by using different mutants, such as dnc, rut, PKA, or amn, lead to centrophobism defect. Moreover, we found that the P[GAL4]C316 flies, used to rescue the amn mutant phenotype, like those previously reported for the learning and memory defect, are severely disturbed in centrophobism behavior. Remarkably, those flies are perfectly randomly distributed in the arena, suggesting that C316 flies carry an important mutated-gene implicated in neuronal networks required for proper spatial orientation.