Sivertsen MS, Glover JC, Perreault MC. Organization of pontine reticulospinal inputs to motoneurons controlling axial and limb muscles in the neonatal mouse. J Neurophysiol 112: 1628 -1643, 2014. First published June 18, 2014 doi:10.1152/jn.00820.2013.-Using optical recording of synaptically mediated calcium transients and selective spinal lesions, we investigated the pattern of activation of spinal motoneurons (MNs) by the pontine reticulospinal projection in isolated brain stem-spinal cord preparations from the neonatal mouse. Stimulation sites throughout the region where the pontine reticulospinal neurons reside reliably activated MNs at cervical, thoracic, and lumbar levels. Activation was similar in MNs ipsi-and contralateral to the stimulation site, similar in medial and lateral motor columns that contain trunk and limb MNs, respectively, and similar in the L2 and L5 segments that predominantly contain flexor and extensor MNs, respectively. In nonlesioned preparations, responses in both ipsi-and contralateral MNs followed individual stimuli in stimulus trains nearly one-to-one (with few failures). After unilateral hemisection at C1 on the same side as the stimulation, responses had substantially smaller magnitudes and longer latencies and no longer followed individual stimuli. After unilateral hemisection at C1 on the side opposite to the stimulation, the responses were also smaller, but their latencies were not affected. Thus we distinguish two pontine reticulospinal pathways to spinal MNs, one uncrossed and the other crossed, of which the uncrossed pathway transmits more faithfully and appears to be more direct. motor control; descending pathways; brain stem; spinal cord; trunk; limb THE MAMMALIAN RETICULOSPINAL system, consisting of the mesencephalic, pontine, and medullary reticulospinal pathways, is crucial for the control of skeletal musculature (Baker 2011;Kuypers 1964;Lemon 2008;Perreault and Glover 2013;Peterson 1979;Pettersson et al. 2007), but the neural mechanisms by which it initiates and regulates movement are far from understood.Electrical stimulation has been a key method in studying the organization of the mammalian reticulospinal system, revealing excitatory reticulospinal connections (mono-and polysynaptic) with axial motoneurons (MNs), proximal and distal limb MNs, and digit MNs in a variety of species (monkey: Davidson and Buford 2006;Riddle et al. 2009; cat: Drew and Rossignol 1990b;Galea et al. 2010;Grillner et al. 1968;Jankowska et al. 2003;Lloyd 1941;Peterson et al. 1979;Sprague and Chambers 1954;Wilson and Yoshida 1969; rat: Bolzoni et al. 2013; Floeter and Lev-Tov 1993;Umeda et al. 2010; and mouse: Alstermark and Ogawa 2004;Szokol et al. 2008). However, many studies have not differentiated among different areas of the reticular formation (RF) that project to the spinal cord. For example, stimulation of the medial longitudinal fasciculus (MLF), in which many (but certainly not all) reticulospinal axons project, is often used as a proxy for stimulating reticulospinal projectio...