<p>Targeted muscle reinnervation surgery reroutes residual nerve signals into spare muscles, enabling the recovery of neural information through electromyography (EMG). However, EMG signals are often overlapping, making the interpretation of limb functions complicated. Regenerative peripheral nerve interfaces surgically partition the nerve into individual fascicles that reinnervate specific muscle grafts, isolating distinct neural sources for precise control and interpretation of EMG signals. Here we combine targeted muscle reinnervation surgery of polyvalent nerves with a high-density microelectrode array implanted at a single site within a reinnervated muscle, and via mathematical source separation methods, we separate all neural signals that are redirected into a single muscle. In participants with upper-limb amputation, the deconvolution of EMG signals from four reinnervated muscles into motor unit spike trains revealed distinct clusters of motor neurons associated with diverse functional tasks. Our method enabled the extraction of multiple neural commands within a single reinnervated muscle, eliminating the need for surgical nerve division. This approach holds promises for enhancing control over prosthetic limbs and for understanding how the central nervous system encodes movement after reinnervation.</p>