Abstract This study examined the effects of chronic hypoxia (CH) and mid-brain transection on central respiratory-related pH/CO 2 chemosensitivity in cane toads ( Bufo marinus). Toads were exposed to 10 days of CH (10% O 2) following which in vitro brainstem-spinal cord preparations, with the mid-brain attached, were used to examine central pH/CO 2 chemosensitivity. A reduction in artificial cerebral spinal fluid (aCSF) pH increased fictive breathing frequency (fR) and total fictive ventilation. CH reduced fictive fR and total fictive ventilation, compared to controls. Mid-brain transection caused an increase in fictive fR, at the lower aCSF pH levels, in both control and CH preparations. In the CH preparations, mid-brain transection restored fictive breathing to control levels. In both groups, mid-brain transection eliminated fictive breath clustering. The data indicate that CH attenuates central pH/CO 2-sensitive fictive breathing but a mid-brain transection in the middle of the optic lobes abolishes this attenuation. The results suggest that CH induces inhibition of central pH/CO 2 chemoreceptor function via descending inputs from the mid-brain region.