Miriam Kron

Development of adaptive behaviour of the respiratory network: implications for the pontine Kolliker-Fuse nucleus.

Breathing is constantly modulated by afferent sensory inputs in order to adapt to changes in behaviour and environment. The pontine respiratory group, in particular the Kolliker-Fuse nucleus, might be a key structure for adaptive behaviours of the respiratory network. Here, we review the anatomical connectivity of the Kolliker-Fuse nucleus with primary sensory structures and with the medullary respiratory centres and focus on the importance of pontine and medullary postinspiratory neurones in the mediation of respiratory reflexes. Furthermore, we will summarise recent findings from our group regarding ontogenetic changes of respiratory reflexes (e.g., the diving response) and provide evidence that immaturity of the Kolliker-Fuse nucleus might account in neonates for a lack of plasticity in sensory evoked modulations of respiratory activity. We propose that a subpopulation of neurones within the Kolliker-Fuse nucleus represent command neurones for sensory processing which are capable of initiating adaptive behaviour in the respiratory network. Recent data from our laboratory suggest that these command neurones undergo substantial postnatal maturation.

Activation of Orexin B receptors in the pontine Kölliker-Fuse nucleus modulates pre-inspiratory hypoglossal motor activity in rat

Orexins (splice variants A and B) are hypothalamic neuropeptides that have essential functions in control of arousal and nutrition. Lack of Orexins is strongly associated with narcolepsy and sleep disordered breathing. However, the role of Orexins and particularly that of Orexin-B (OXB), in respiratory centres controlling upper-airway patency are less defined. In the present study we performed microinjections of OXB into the pontine Kölliker-Fuse nucleus (KF) of the dorsolateral pons, since this nucleus is particularly involved in the pre-motor control of upper airway muscles. The OXB mediated effects on heart, phrenic (PNA) and hypoglossal (XII-A) nerve activities were analysed in an in situ perfused brainstem preparation. Injection of OXB into the KF evoked significant augmentation of the respiratory frequency. Importantly, OXB provoked particularly prolonged pre-inspiratory discharge of the XII nerve, while no cardiovascular response was observed after KF microinjections. In summary, OXB in the KF exerts an excitatory effect on XII pre-motoneurones. Since pre-inspiratory activity of the XII is important for the decrease in upper airway resistance during inspiration, we conclude that OXB release in the KF has strong implications in the state-dependent control of upper airway patency under physiological and pathophysiological conditions.

Emergence of brain-derived neurotrophic factor-induced postsynaptic potentiation of NMDA currents during the postnatal maturation of the Kölliker–Fuse nucleus of rat

The Kölliker–Fuse nucleus (KF) contributes essentially to respiratory pattern formation and adaptation of breathing to afferent information. Systems physiology suggests that these KF functions depend on NMDA receptors (NMDA‐R). Recent investigations revealed postnatal changes in the modulation of glutamatergic neurotransmission by brain‐derived neurotrophic factor (BDNF) in the KF. Therefore, we investigated postnatal changes in NMDA‐R subunit composition and postsynaptic modulation of NMDA‐R‐mediated currents by BDNF in KF slice preparations derived from three age groups (neonatal: postnatal day (P) 1–5; intermediate: P6–13; juvenile: P14–21). Immunohistochemistry showed a developmental up‐regulation of the NR2D subunit. This correlated with a developmental increase in decay time of NMDA currents and a decline of desensitization in response to repetitive exogenous NMDA applications. Thus, developmental up‐regulation of the NR2D subunit, which reduces the Mg2+ block of NMDA‐R, causes these specific changes in NMDA current characteristics. This may determine the NMDA‐R‐dependent function of the mature KF in the control of respiratory phase transition. Subsequent experiments revealed that bath‐application of BDNF progressively potentiated these repetitively evoked NMDA currents only in intermediate and juvenile age groups. Pharmacological inhibition of protein kinase C (PKC), as a downstream component of the BDNF–tyrosine kinase B receptor (trkB) signalling, prevented BDNF‐induced potentiation of NMDA currents. BDNF‐induced potentiation of NMDA currents in later developmental stages might be essential for synaptic plasticity during the adaptation of the breathing pattern in response to peripheral/central commands. The lack of plasticity in neonatal neurones strengthens the hypothesis that the respiratory network becomes permissive for activity‐dependent plasticity with ongoing postnatal development.

Infant brain stem is prone to the generation of spreading depression during severe hypoxia.

Spreading depression (SD) resembles a concerted, massive neuronal/glial depolarization propagating within the gray matter. Being associated with cerebropathology, such as cerebral ischemia or hemorrhage, epileptic seizures, and migraine, it is well studied in cortex and hippocampus. We have now analyzed the susceptibility of rat brain stem to hypoxia-induced spreading depression-like depolarization (HSD), which could critically interfere with cardiorespiratory control. In rat brain stem slices, severe hypoxia (oxygen withdrawal) triggered HSD within minutes. The sudden extracellular DC potential shift of approximately -20 mV showed the typical profile known from other brain regions and was accompanied by an intrinsic optical signal (IOS). Spatiotemporal IOS analysis revealed that in infant brain stem, HSD was preferably ignited within the spinal trigeminal nucleus and then mostly spread out medially, invading the hypoglossal nucleus, the nucleus of the solitary tract (NTS), and the ventral respiratory group (VRG). The neuronal hypoxic depolarizations underlying the generation of HSD were massive, but incomplete. The propagation velocity of HSD and the associated extracellular K(+) rise were also less marked than in other brain regions. In adult brain stem, HSD was mostly confined to the NTS and its occurrence was facilitated by hypotonic solutions, but not by glial poisoning or block of GABAergic and glycinergic synapses. In conclusion, brain stem tissue reliably generates propagating HSD episodes, which may be of interest for basilar-type migraine and brain stem infarcts. The preferred occurrence of HSD in the infant brain stem and its propagation into the VRG may be of importance for neonatal brain stem pathology such as sudden infant death syndrome.

Developmental changes in brain-derived neurotrophic factor-mediated modulations of synaptic activities in the pontine Kölliker-Fuse nucleus of the rat

The Kölliker–Fuse nucleus (KF), part of the respiratory network, is involved in the modulation of respiratory phase durations in response to peripheral and central afferent inputs. The KF is immature at birth. Developmental changes in its physiological and anatomical properties have yet to be investigated. Since brain‐derived neurotrophic factor (BDNF) is of major importance for the maturation of neuronal networks, we investigated its effects on developmental changes in the KF on different postnatal days (neonatal, P1–5; intermediate, P6–13; juvenile, P14–21) by analysing single neurones in the in vitro slice preparation and network activities in the perfused brainstem preparation in situ. The BDNF had only weak effects on the frequency of mixed excitatory and inhibitory spontaneous postsynaptic currents (sPSCs) in neonatal slice preparations. Postnatally, in the intermediate and juvenile age groups, a significant augmentation of the sPSC frequency was observed in the presence of 100 pm BDNF (+23.5 ± 12.6 and +76.7 ± 28.4%, respectively). Subsequent analyses of BDNF effects on evoked excitatory postsynaptic currents (eEPSCs) revealed significant enhancement of eEPSC amplitude of +20.8 ± 7.0% only in juvenile stages (intermediates, −13.2 ± 4.8%). On the network level, significant modulation of phrenic nerve activity following BDNF microinjection into the KF was also observed only in juveniles. The data suggest that KF neurones are subject to BDNF‐mediated fast synaptic modulation after completion of postnatal maturation. After maturation, BDNF contributes to modulation of fast excitatory neurotransmission in respiratory‐related KF neurones. This may be important for network plasticity associated with the processing of afferent information.

Postnatal emergence of synaptic plasticity associated with dynamic adaptation of the respiratory motor pattern.

The shape of the three-phase respiratory motor pattern (inspiration, postinspiration, late expiration) is controlled by a central pattern generator (CPG) located in the ponto-medullary brainstem. Synaptic interactions between and within specific sub-compartments of the CPG are subject of intensive research. This review addresses the neural control of postinspiratory activity as the essential determinant of inspiratory/expiratory phase duration. The generation of the postinspiratory phase depends on synaptic interaction between neurones of the nucleus tractus solitarii (NTS), which relay afferent inputs from pulmonary stretch receptors, and the pontine Kölliker-Fuse nucleus (KF) as integral parts of the CPG. Both regions undergo significant changes during the first three postnatal weeks in rodents. Developmental changes in glutamatergic synaptic functions and its modulation by brain-derived neurotrophic factor may have implications in synaptic plasticity within the NTS/KF axis. We propose that dependent on these developmental changes, the CPG becomes permissive for short- and long-term plasticity associated with environmental, metabolic and behavioural adaptation of the breathing pattern.

Developmental changes in the BDNF-induced modulation of inhibitory synaptic transmission in the Kölliker–Fuse nucleus of rat

The Kölliker–Fuse nucleus (KF), part of the pontine respiratory group, is involved in the control of respiratory phase duration, and receives both excitatory and inhibitory afferent input from various other brain regions. There is evidence for developmental changes in the modulation of excitatory inputs to the KF by the neurotrophin brain‐derived neurotrophic factor (BDNF). In the present study we investigated if BDNF exerts developmental effects on inhibitory synaptic transmission in the KF.

Expression and Function of Serotonin 2A and 2B Receptors in the Mammalian Respiratory Network

Neurons of the respiratory network in the lower brainstem express a variety of serotonin receptors (5-HTRs) that act primarily through adenylyl cyclase. However, there is one receptor family including 5-HT2A, 5-HT2B, and 5-HT2C receptors that are directed towards protein kinase C (PKC). In contrast to 5-HT2ARs, expression and function of 5-HT2BRs within the respiratory network are still unclear. 5-HT2BR utilizes a Gq-mediated signaling cascade involving calcium and leading to activation of phospholipase C and IP3/DAG pathways. Based on previous studies, this signal pathway appears to mediate excitatory actions on respiration. In the present study, we analyzed receptor expression in pontine and medullary regions of the respiratory network both at the transcriptional and translational level using quantitative RT-PCR and self-made as well as commercially available antibodies, respectively. In addition we measured effects of selective agonists and antagonists for 5-HT2ARs and 5-HT2BRs given intra-arterially on phrenic nerve discharges in juvenile rats using the perfused brainstem preparation. The drugs caused significant changes in discharge activity. Co-administration of both agonists revealed a dominance of the 5-HT2BR. Given the nature of the signaling pathways, we investigated whether intracellular calcium may explain effects observed in the respiratory network. Taken together, the results of this study suggest a significant role of both receptors in respiratory network modulation.

Postnatal Changes in Morphology and Dendritic Organization of Neurones Located in the Area of the Kölliker-Fuse Nucleus of Rat

The Kölliker-Fuse nucleus (KF) is an integral part of the central pattern generator for breathing and shows postnatal development of synaptic functions and cyto-architectural structure. Here, we analyzed the postnatal changes in cell morphology of biocytin-labelled KF neurones. Developmental analyses revealed an increasing size of somas and dendritic length. These changes were accompanied by changes in the orientation of the main dendritic branches from a diffuse orientation in neonates to a predominant medio-lateral orientation in juveniles. These developmental changes may allow for synaptic contacts with multiple ascending fibre tracts required for the processing of multi-modal respiratory inputs in the KF.