The present study was performed to establish an anatomical context for studies of phrenic motor function in mouse models of central nervous system trauma and disease. Application of cholera toxin β-subunit to the diaphragm of adult C57BL/6 mice revealed a columnar organization of phrenic motoneurons (PhMNs) which extended from rostral C3 to C6. Injection of Miniruby into the ventrolateral medulla revealed decussating, anterogradely labeled axons in the cervical spinal cord. In addition, application of the transneuronal tracer pseudorabies virus (PRV) to the right hemidiaphragm demonstrated a population of putative pre-phrenic interneurons at the level of the infected PhMN pool. These neuroanatomical features of the mouse phrenic nucleus are consistent with those described in other species and provide a foundation for studies of neuroplasticity and repair in relation to a functionally and anatomically identified spinal network.
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