Spontaneous limb preferences exist in numerous species. To investigate the underlying mechanisms of these preferences, different methods, such as training, have been developed to shift preferences artificially. However, studies that systematically examine the effects of shifting preferences on behaviour and physiology are largely missing. Therefore, the aim of this study was to assess the impact of shifting paw preferences via training on spontaneous home cage behaviour, as well as anxiety-like behaviour and exploratory locomotion (Elevated plus maze test, Dark light test, Open field test, Free exploration test), learning performance (Labyrinth-maze) and stress hormones (fecal corticosterone metabolites) in laboratory mice (Mus musculus f. domestica). For this, we assessed spontaneous paw preferences of C57BL/6J females (Nambilateral = 23, Nleft = 23, Nright = 25). Subsequently, half of the individuals from each category were trained once a week for four weeks in a food-reaching task to use either their left or right paw, respectively, resulting in six groups: AL, AR, LL, LR, RL, RR. After training, a battery of behavioural tests was performed and spontaneous preferences were assessed again. Our results indicate that most mice were successfully trained and the effect of training was present days after training. However, a significant difference of preferences between RL and LL mice during training suggests a rather low training success of RL mice. Additionally, preferences of L mice differed from those of A and R mice after training, indicating differential long-term effects of training in these groups. Furthermore, left paw training led to higher levels of self-grooming, possibly as a displacement behaviour, and more time spent in the light compartment of the Dark light test. However, overall, there was no systematic influence of training on behavioural measures and stress hormones. Different explanations for this lack of influence, such as the link between training and hemispheric functioning or the intensity and ecological relevance of the training, are discussed.
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