Staudenmann D, van Dieën JH, Stegeman DF, Enoka RM. Increase in heterogeneity of biceps brachii activation during isometric submaximal fatiguing contractions: a multichannel surface EMG study. J Neurophysiol 111: 984 -990, 2014. First published December 11, 2013 doi:10.1152/jn.00354.2013.-The effects of fatigue emerge from the beginning of sustained submaximal contractions, as shown by an increase in the amplitude of the surface electromyogram (EMG). The increase in EMG amplitude is attributed to an augmentation of the excitatory drive to the motor neuron pool that, more importantly than increasing discharge rates, recruits additional motor units for the contraction. The aim of this study was to determine whether the spatiotemporal distribution of biceps brachii (BB) activity becomes more or less heterogeneous during a fatiguing isometric contraction sustained at a submaximal target force. Multiple electrodes were attached over the entire BB muscle, and principal component analysis (PCA) was used to extract the representative information from multiple monopolar EMG channels. The development of heterogeneity during the fatiguing contraction was quantified by applying a cluster algorithm on the PCA-processed EMG amplitudes. As shown previously, the overall EMG amplitude increased during the sustained contraction, whereas there was no change in coactivation of triceps brachii. However, EMG amplitude did not increase in all channels and even decreased in some. The change in spatial distribution of muscle activity varied across subjects. As found in other studies, the spatial distribution of EMG activity changed during the sustained contraction, but the grouping and size of the clusters did not change. This study showed for the first time that muscle activation became more heterogeneous during a sustained contraction, presumably due to a decrease in the strength of common inputs with the recruitment of additional motor units.