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Ultrastructural localization of the binding fragment of tetanus toxin in putative γ‐aminobutyric acidergic terminals in the intermediolateral cell column: A potential basis for sympathetic dysfunction in generalized tetanus

Tetanus toxin (TeTx) causes sympathetic hyperactivity, a major cause of mortality in generalized tetanus, apparently by obstructing the inhibition of sympathetic preganglionic neurons (SPNs). Neuroanatomic tracing and immunohistochemistry were used to investigate whether axon terminals in the intermediolateral cell column (IML) that synapse on SPNs and use the inhibitory neurotransmitter γ‐aminobutyric acid (GABA) may be infected transsynaptically with TeTx. The binding fragment of TeTx (TTC; an atoxic surrogate of TeTx) and the cholera toxin B subunit (CTB; a retrograde tracer) were injected into the rat superior cervical ganglion and, over 16–48 hours, were transported to the ipsilateral IML in the caudal half of the last cervical and first three thoracic spinal cord segments. With light microscopy, diffuse CTB immunolabeling extended throughout SPN perikarya and dendrites. Punctate TTC and GABA immunolabeling were accumulated densely in the neuropil between and surrounding SPN processes. With electron microscopy, 54% of the axon terminals in the IML (n = 1,337 terminals) were TTC immunolabeled (TTC+), and 25% contained putative neurotransmitter levels of GABA immunolabeling (GABA+). On average, GABA+ terminals had a 76% chance of also being TTC+ and a 62% greater chance of being TTC+ than GABA− terminals (P < 0.000001). Axon terminals were just as likely to be TTC+ and/or GABA+ regardless of whether the dendrites they synapsed on were large (>1 μM) or small in cross‐sectional area or were labeled retrogradely. Sympathetic hyperactivity in tetanus may involve 1) retrograde and transsynaptic transport of TeTx by SPNs and 2) at least in part, an infection of GABAergic terminals in the IML. J. Comp. Neurol. 419:471–484, 2000. © 2000 Wiley‐Liss, Inc.