The Kölliker-Fuse nucleus orchestrates the timing of expiratory abdominal nerve bursting.

Coordination of respiratory pump and valve muscle activity is essential for normal breathing. A hallmark respiratory response to hypercapnia and hypoxia is the emergence of active exhalation, characterized by abdominal muscle pumping during the late one-third of expiration (late-E phase). Late-E abdominal activity during hypercapnia has been attributed to the activation of expiratory neurons located within the parafacial respiratory group (pFRG). However, the mechanisms that control emergence of active exhalation, and its silencing in restful breathing, are not completely understood. We hypothesized that inputs from the Kölliker-Fuse nucleus (KF) control the emergence of late-E activity during hypercapnia. Previously, we reported that reversible inhibition of the KF reduced postinspiratory (post-I) motor output to laryngeal adductor muscles and brought forward the onset of hypercapnia-induced late-E abdominal activity. Here we explored the contribution of the KF for late-E abdominal recruitment during hypercapnia by pharmacologically disinhibiting the KF in in situ decerebrate arterially perfused rat preparations. These data were combined with previous results and incorporated into a computational model of the respiratory central pattern generator. Disinhibition of the KF through local parenchymal microinjections of gabazine (GABAA receptor antagonist) prolonged vagal post-I activity and inhibited late-E abdominal output during hypercapnia. In silico, we reproduced this behavior and predicted a mechanism in which the KF provides excitatory drive to post-I inhibitory neurons, which in turn inhibit late-E neurons of the pFRG. Although the exact mechanism proposed by the model requires testing, our data confirm that the KF modulates the formation of late-E abdominal activity during hypercapnia. NEW & NOTEWORTHY The pons is essential for the formation of the three-phase respiratory pattern, controlling the inspiratory-expiratory phase transition. We provide functional evidence of a novel role for the Kölliker-Fuse nucleus (KF) controlling the emergence of abdominal expiratory bursts during active expiration. A computational model of the respiratory central pattern generator predicts a possible mechanism by which the KF interacts indirectly with the parafacial respiratory group and exerts an inhibitory effect on the expiratory conditional oscillator.

The Kölliker-Fuse nucleus acts as a timekeeper for late-expiratory abdominal activity.

While the transition from the inspiratory to the post-inspiratory (post-I) phase is dependent on the pons, little attention has been paid to understanding the role of the pontine respiratory nuclei, specifically the Kölliker-Fuse nucleus (KF), in transitioning from post-I to the late expiratory (late-E) activity seen with elevated respiratory drive. To elucidate this, we used the in situ working heart-brainstem preparation of juvenile male Holtzman rats and recorded from the vagus (cVN), phrenic (PN) and abdominal nerves (AbN) during baseline conditions and during chemoreflex activation [with potassium cyanide (KCN; n=13) or hypercapnia (8% CO2; n=10)] to recruit active expiration. Chemoreflex activation with KCN increased PN frequency and cVN post-I and AbN activities. The inhibition of KF with isoguvacine microinjections (10mM) attenuated the typical increase in PN frequency and cVN post-I activity, and amplified the AbN response. During hypercapnia, AbN late-E activity emerged in association with a significant reduction in expiratory time. KF inhibition during hypercapnia significantly decreased PN frequency and reduced the duration and amplitude of post-I cVN activity, while the onset of the AbN late-E bursts occurred significantly earlier. Our data reveal a negative relationship between KF-induced post-I and AbN late-E activities, suggesting that the KF coordinates the transition between post-I to late-E activity during conditions of elevated respiratory drive.

Oxygen in demand: How oxygen has shaped vertebrate physiology.

In response to varying environmental and physiological challenges, vertebrates have evolved complex and often overlapping systems. These systems detect changes in environmental oxygen availability and respond by increasing oxygen supply to the tissues and/or by decreasing oxygen demand at the cellular level. This suite of responses is termed the oxygen transport cascade and is comprised of several components. These components include 1) chemosensory detectors that sense changes in oxygen, carbon dioxide, and pH in the blood, and initiate changes in 2) ventilation and 3) cardiac work, thereby altering the rate of oxygen delivery to, and carbon dioxide clearance from, the tissues. In addition, changes in 4) cellular and systemic metabolism alters tissue-level metabolic demand. Thus the need for oxygen can be managed locally when increasing oxygen supply is not sufficient or possible. Together, these mechanisms provide a spectrum of responses that facilitate the maintenance of systemic oxygen homeostasis in the face of environmental hypoxia or physiological oxygen depletion (i.e. due to exercise or disease). Bill Milsom has dedicated his career to the study of these responses across phylogenies, repeatedly demonstrating the power of applying the comparative approach to physiological questions. The focus of this review is to discuss the anatomy, signalling pathways, and mechanics of each step of the oxygen transport cascade from the perspective of a Milsomite. That is, by taking into account the developmental, physiological, and evolutionary components of questions related to oxygen transport. We also highlight examples of some of the remarkable species that have captured Bill's attention through their unique adaptations in multiple components of the oxygen transport cascade, which allow them to achieve astounding physiological feats. Bill's research examining the oxygen transport cascade has provided important insight and leadership to the study of the diverse suite of adaptations that maintain cellular oxygen content across vertebrate taxa, which underscores the value of the comparative approach to the study of physiological systems.

Expiration: breathing's other face.

The evolution of the aspiration pump seen in tetrapod vertebrates from the buccal-pharyngeal force pump seen in air breathing fish and amphibians appears to have first involved the production of active expiration. Active inspiration arose later. This appears to have involved reconfiguration of a parafacial oscillator (now the parafacial respiratory group/retrotrapezoid nucleus (pFRG/RTN)) to produce active expiration, followed by reconfiguration of a paravagal oscillator (now the preBötC) to produce active inspiration. In the ancestral breathing cycle, inspiration follows expiration, which is in turn followed by glottal closure and breath holding. When both rhythms are expressed, as they are in reptiles and birds, and mammals under conditions of elevated respiratory drive, the pFRG/RTN appears to initiate the respiratory cycle. We propose that the coordinated output of this system is a ventilation cycle characterized by four phases. In reptiles, these consist of active inspiration (I), glottal closure (E1), a pause (an apnea or breath hold) (E2), and an active expiration (E3) that initiates the next cycle. In mammals under resting conditions, active expiration (E3) is suppressed and inspiration (I) is followed by airway constriction and diaphragmatic braking (E1) (rather than glottal closure) and a short pause at end-expiration (E2). As respiratory drive increases in mammals, expiratory muscle activity appears. Frequently, it first appears immediately preceding inspiration (E3) just as it does in reptiles. It can also appear in E1, however, and it is not yet clear what mechanisms underlie when and where in the cycle it appears. This may reflect whether the active expiration is recruited to enhance tidal volume, increase breathing frequency, or both.

Visual discrimination learning in the fire-bellied toad Bombina orientalis.

This study explored the visual discrimination learning ability of fire-bellied toads (Bombina orientalis). Two groups of toads were trained in a simultaneous visual discrimination task involving video footage of either black crickets on a white background (black-cricket toads) or white crickets on a black background (white-cricket toads). Fifteen widely spaced acquisition trials were followed by 12 reversal trials. Successful learning was observed by decreased incorrect snapping and reduced latency to snap at the correct stimulus (S+) during acquisition; however, white-cricket toads executed significantly more incorrect snaps than did black-cricket toads. Both groups of toads could master the reversal task as measured by latency to snap at S+, but not as measured by the proportion of incorrect snaps. Despite the stronger potency of the black-cricket stimulus, the results showed that toads can learn a simultaneous discrimination task and a reversal of its contingency. This elaborate form of learning appears to be conserved among vertebrates.