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The Journal of Neuroscience, June 1, 2003, 23(11):4657-4666

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Brainstem Substrates of Sympatho-Motor Circuitry Identified Using Trans-Synaptic Tracing with Pseudorabies Virus Recombinants

Ilan A. Kerman,^1,2,3 Lynn W. Enquist,⁴ Stanley J. Watson,³ and Bill J. Yates^1,2

¹ Department of Neuroscience University of Pittsburgh, Pittsburgh, Pennsylvania 15213, ² Department of Otolaryngology, University of Pittsburgh, Pittsburgh, Pennsylvania 15213, ³ Mental Health Research Institute, Department of Psychiatry, University of Michigan, Ann Arbor, Michigan 48109, and ⁴ Department of Molecular Biology, Princeton University, Princeton New Jersey 08544

Previous physiological investigations have suggested the existence of a neural circuit that coordinates activation of motor and autonomic efferents before or at the onset of exercise. Traditionally these circuits have been postulated to involve forebrain areas. However, overlapping populations of medullary reticular formation neurons that participate in motor or autonomic control have been described previously, suggesting that individual pontomedullary reticular formation neurons may coordinate both motor and autonomic responses. We tested this hypothesis by conducting transneuronal retrograde tracing of motor and sympathetic nervous system pathways in rats using recombinant strains of pseudorabies virus (PRV). A PRV strain expressing the green fluorescent protein (PRV-152) was injected into the left gastrocnemius muscle, which was surgically sympathectomized, whereas another recombinant (PRV-BaBlu) was injected into the left adrenal gland. Immunofluorescence methods using monospecific antisera and distinct fluorophores identified neurons infected with one or both of the recombinants. Brainstem neurons coinfected with both PRV recombinants, which presumably had collateralized projections to both adrenal sympathetic preganglionic neurons and gastrocnemius motoneurons, were observed in several areas of the pontomedullary reticular formation. The largest number of such neurons was located in the rostral ventromedial medulla within the ventral gigantocellular nucleus, gigantocellular nucleus pars alpha, raphe obscurus, and raphe magnus. These neurons are candidates for relaying central command signals to the spinal cord.

Key words: pseudorabies virus; rats; dual-labeling immunofluorescence; central command; stress; exercise; motor pathway; sympathetic nervous system; raphe nuclei; pontomedullary reticular formation; rostral ventromedial medulla

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Received Dec. 26, 2002; revised Mar. 17, 2003; accepted Mar. 17, 2003.

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