The inspiratory-related inhibition does not arise from structures rostral to the pre-Bötzinger Complex.A, Sagittal view of the brainstem showing levels of transection (dashed lines labeled 1through 6) taken serially, in relation to organization of ventrolateral brainstem respiratory nuclei.B, Time course of the effects on ∫C4 inspiratory burst amplitude (in volts) and burst frequency (F) of locally applying bicuculline (60 sec, 1 mm) over the C4 PMN pool after transection of the brainstem at levels 1(Bi) and 5 (Bii). Individual data points represent amplitude and frequency of single inspiratory cycles. The line in the top panel ofBi represents the moving average of the C4 burst amplitude response to bicuculline applied after transection at level1. This moving average is included in Biito illustrate that the effects of bicuculline were unaffected with progressively more caudal transections up to level 5. Removal of the rostral component of the pre-Bötzinger Complex with transection at level 6 disrupted both respiratory rhythm and C4 burst amplitude (Biii), precluding further analysis of responses to bicuculline. BötC, Bötzinger Complex; VII, facial nucleus;XII, hypoglossal nucleus; cNA, compact division of nucleus ambiguus; LRN, lateral reticular nucleus; NA, nucleus ambiguus;Pre-BötC, pre-Bötzinger Complex;rVRG, rostral–ventral respiratory group).