For each experimental session a new word list was presented. The list was composed of complexity-matched words (see Supporting Information). During the mental activity, subjects were instructed to imagine the movements from a first person perspective and to employ kinesthetic cues (e.g. the feeling of the pen in their hand). The anodal tDCS was administered for 13 min during the whole course of the MP. Continuous direct currents were transferred by saline-soaked surface sponge electrodes (surface 20 cm2) and delivered by a clinical microcurrent stimulator (Soterix, USA) with a maximum output of 2 mA. Five different electrode montages
were tested to find the optimal position for DC stimulation in increasing the neuroplastic effects of mental imagery on motor
performance. The excitatory tDCS was applied over the: (i) right AZD2014 solubility dmso M1, (ii) right premotor area (PMA), (iii) right SMA, (iv) right cerebellar hemisphere, and (v) left dorsolateral prefrontal cortex. For M1 tDCS, the anode electrode was positioned above C3 (international 10-20 system) (Nitsche et al., 2003b). For stimulation of the premotor cortex, it was moved 2 cm forward and 2 cm to the midline relative to the M1 position (Nitsche et al., 2003b). The SMA tDCS was performed with the anode electrode placed 2 cm anterior to the vertex (position Cz), in the sagittal midline (Cunnington et al., 1996). For DC stimulation of the dorsolateral prefrontal cortex, the anode electrode was positioned 5 cm forward relative to C3 (Nitsche et al., 2003b). In all cases, the reference electrode was placed above the contralateral orbit. For cerebellar tDCS, electrodes were placed with GSK2118436 solubility dmso one (anode electrode) over the right cerebellar hemisphere, 3 cm lateral to the inion (Ugawa et al., 1995), and the other over the deltoid muscle (Ferrucci et al., 2008). These methods of electrode montage have been used in previous studies and been shown to be effective in the modulation of cerebral activity. The order of stimulation condition was counterbalanced across subjects. The anodal tDCS was administered with a current strength of 2 mA. In
the sham session, tDCS was applied over the M1 for 30 s, Vitamin B12 a method shown to achieve a good level of blinding (Gandiga et al., 2006). In each experimental session, motor performance was assessed by the handwriting test. This test measured legibility and writing time, important elements in handwriting performance (Bonney, 1992). Handwriting is a complex perceptual–motor skill that includes fine motor control (hand manipulation, bilateral integration, and motor planning) (Feder & Majnemer, 2007). For the test, the subjects were instructed to copy a six-word set with the non-dominant hand on a blank sheet of paper positioned on a table to the left of the subject. The word list was presented approximately three inches away from the paper. The handwriting task was performed with spontaneous production, free from the influence of the writing instructions.