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A complex propriospinal network is synaptically coupled to phrenic and intercostal motoneurons, and this makes it difficult to predict how gray matter intraspinal microstimulation (ISMS) will recruit respiratory motor units. We therefore mapped the cervical and high thoracic gray matter at locations which ISMS activates diaphragm (DIA) and external intercostal (EIC) motor units. Respiratory muscle electromyography (EMG) was recorded in anesthetized female spinally intact adult rats while a stimulating electrode was advanced ventrally into the spinal cord in 600 μm increments. At each depth, single biphasic stimuli were delivered at 10–90 μA during both the inspiratory and expiratory phase independently. Twenty electrode tracks were made from C2-T1 at medial and lateral gray matter locations. During inspiration, ISMS evoked DIA and EIC activity throughout C2-T1 gray matter locations, with mutual activation occurring at 17 ± 9% of sites. During inspiratory phase ISMS the average latency for DIA activation was 4.40 ± 0.70 ms. During the expiratory phase, ISMS-induced DIA activation required electrodes to be in close proximity to the phrenic motoneuron pool, and average activation latency was 3.30 ± 0.50 ms. We conclude that appropriately targeted ISMS can co-activate DIA and EIC motor units, and endogenous respiratory drive has a powerful impact on ISMS-induced respiratory motor unit activation. The long latency diaphragm motor unit activation suggests the presence of a complex propriospinal network that can modulate phrenic motor output.Intraspinal microstimulation (ISMS) co-activates respiratory muscles.Inspiratory triggered ISMS is most effective at activating respiratory muscles.ISMS primarily activates the diaphragm with latencies > 2.5 ms.ISMS is promising for activating respiratory circuits for therapeutic applications.