Spinal Interneurons With Dual Axon Projections to Knee-Extensor and Hip-Extensor Motor Pools

Abstract

The central nervous system (CNS) may simplify control of limb movements by activating certain combinations of muscles together, i.e., muscle synergies. Little is known, however, about the spinal cord interneurons that activate muscle synergies by exciting sets of motoneurons for different muscles. The turtle spinal cord, even without brain inputs and movement-related sensory feedback, can generate the patterns of motoneuron activity underlying forward swimming, three forms of scratching, and limb withdrawal. Spinal interneurons activated during scratching are typically activated during all three forms of scratching, to different degrees, even though each form of scratching has its own knee-hip synergy. Such spinal interneurons are also typically activated rhythmically during scratching motor patterns, with hip-related timing. We proposed a hypothesis that such interneurons that are most active during rostral scratch stimulation project their axons to both knee-extensor and hip-flexor motoneurons, thus generating the rostral scratch knee-hip synergy, while those interneurons most active during pocket scratch stimulation project their axons to both knee-extensor and hip-extensor motoneurons, thus generating the pocket scratch knee-hip synergy. The activity of the entire population would then generate the appropriate synergy, depending on the location of sensory stimulation. Mathematical modeling has demonstrated that this hypothesis is feasible. Here, we provide one test of this hypothesis by injecting two fluorescent retrograde tracers into the regions of knee-extensor motoneurons (more rostrally) and hip-extensor motoneurons (more caudally). We found that there were double-labeled interneurons, which projected their axons to both locations. The dual-projecting interneurons were widely distributed rostrocaudally, dorsoventrally, and mediolaterally within the hindlimb enlargement and pre-enlargement spinal segments examined. The existence of such dual-projecting interneurons is consistent with the hypothesis that they contribute to generating the knee-hip synergy for pocket scratching. The dual-projecting interneurons, however, were only about 1% of the total interneurons projecting to each location, which suggests that they might be one of several contributors to the appropriate knee-hip synergy. Indirect projections to both motor pools and/or knee extensor-dedicated interneurons might also contribute. There is evidence for dual-projecting spinal interneurons in frogs and mice as well, suggesting that they may contribute to limb motor control in a variety of vertebrates.