Asymmetric neuromodulation in the respiratory network contributes to rhythm and pattern generation

Rishi R Dhingra^1,2*Peter M MacFarlane^1,3Peter J Thomas⁴Julian FR Paton⁵Mathias Dutschmann^1,2*

• ¹School of Medicine, Case Western Reserve University, Cleveland, Ohio, United States
• ²Florey Institute of Neuroscience and Mental Health, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, Victoria, Australia
• ³Rainbow Babies & Children's Hospital, Cleveland, Ohio, United States
• ⁴Case Western Reserve University, Cleveland, Ohio, United States
• ⁵The University of Auckland, Auckland, Auckland, New Zealand

Like other brain circuits, the brainstem respiratory network is continually modulated by neurotransmitters that activate slow metabotropic receptors. In many cases, activation of these receptors only subtly modulates the respiratory motor pattern. However, activation of some receptor types evokes the arrest of the respiratory motor pattern as can occur following the activation of µ-opioid receptors. We propose that the varied effects of neuromodulation on the respiratory motor pattern depend on the pattern of neuromodulator receptor expression and their influence on the excitability of their post-synaptic targets. Because a comprehensive characterization of these cellular properties across the respiratory network remains challenging, we test our hypothesis by combining computational modelling with ensemble electrophysiologic recording in the pre-Bötzinger complex (pre-BötC) using high-density multi-electrode arrays (MEA). Our computational model encapsulates the hypothesis that neuromodulatory transmission is organized asymmetrically across the respiratory network to promote rhythm and pattern generation. To test this hypothesis, we increased the strength of subsets of neuromodulatory connections in the model and used selective agonists in situ while monitoring pre-BötC ensemble activities. The in silico simulations of increasing slow inhibition were consistent with experiments examining the effect of systemic administration of the 5HT1aR agonist 8-OH-DPAT. Similarly, the effects of increasing slow excitation in the model were experimentally confirmed in pre-BötC ensemble activities before and after systemic administration of the µ-opioid receptor agonist fentanyl. We conclude that asymmetric neuromodulation can contribute to respiratory rhythm and pattern generation and accounts for its varied effects on breathing.