Substance P Reshapes Inspiratory and Postinspiratory Activity Generated in Horizontal Brainstem Slices

Abstract

The triple oscillator hypothesis posits that breathing is generated by three rhythmogenic microcircuits contained within the ventrolateral medulla. The preBötzinger Complex (preBötC) generates the inexorable inspiratory phase of breathing, whereas the conditional postinspiratory and active expiratory phases of breathing are generated by the Postinspiratory Complex (PiCo) and lateral parafacial respiratory group (PFL), respectively. Neuromodulators including norepinephrine, opioids, and somatostatin are known to differentially modulate the activity of each rhythmic circuit, likely contributing to the independence of each breathing phase and the flexibility of the respiratory rhythm. However, whether the functional boundaries of each of these microcircuits are static or can change dynamically based on the neuromodulatory state of the network is unknown. The neuromodulator Substance P (SP) facilitates breathing frequency in vivo and also in medullary slices that isolate the preBötC in vitro. Using a neonatal mouse horizontal brainstem slice preparation that preserves the rostrocaudal extent of the ventral medulla, we tested the hypothesis that SP differentially modulates bursting activity in the preBötC and PiCo and explored how SP may reshape the distribution of inspiratory activity within the wider medullary network. Rhythmic bursting activity was simultaneously recorded from the preBötC and PiCo (n=5). As expected, recordings from the PiCo, located ~500μm rostral to the preBötC, revealed a slow postinspiratory rhythm with robust bursts occurring after only ~1/12 preBötC bursts under control conditions. Following introduction of 0.5–1.0μM SP into the bath superfusion solution, PiCo bursts decreased in amplitude (~40%) and accelerated such that they occurred 1:1 with preBötC bursts without changing the time interval between peak preBötC and PiCo activity. In some PiCo recordings, we noted a small amount of inspiratory activity synchronized with preBötC bursts that was eliminated in the presence of SP. To test whether SP alters the spatial distribution of inspiratory activity along the ventral medulla, we simultaneously recorded inspiratory population activity generated at the preBötC and 400–600μm rostral (n=8). Following application of 1μM SP frequency was increased (~90%) and inspiratory burst amplitude was reduced by ~20% at the preBötC, whereas burst amplitude decreased by ~50% in rostral recordings. Based on these preliminary experiments, we hypothesize that SP reshapes breathing activity generated within the medulla by facilitating postinspiratory activity and reducing the extent of inspiratory activity rostral of the preBötC to shrink the size of the active inspiratory network. Differential neuromodulatory properties and dynamic functional boundaries of the rhythmogenic circuits that generate breathing may contribute to the remarkable flexibility and robustness of this vital physiological behavior.Support or Funding InformationNIH F32 HL134207NIH R01 HL126523This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.