Pontine GABAergic pathways: role and plasticity in the hypoxic ventilatory response

The hypoxic ventilatory response (HVR) was compared before and after uni- and bi-lateral injections of bicuculline, a GABA(A) receptor antagonist, into the ventrolateral (vl) pons and before and after conditioning animals to chronic sustained hypoxia (CSH). The HVR was assessed by recording phrenic nerve activity (PNA) during and after brief exposures to hypoxia (8% O(2) and 92% N(2) for 45s). Inspiratory (T(I)) and expiratory (T(E)) durations were averaged before hypoxia, at the peak breathing frequency during hypoxia, before the end of hypoxia, immediately after hypoxia, and 60s after hypoxia. Blocking GABA(A) receptors in the vl pons prolonged T(E) during, but not after hypoxia. After CSH induced by 14 days in a hypobaric chamber (0.5atm), the HVR was attenuated compared to that in the naive animals. This plasticity of HVR was associated with selective induction of alpha6 and delta GABA(A) receptor subunit mRNAs specifically in the pons compared to the medulla. These physiological and molecular results illustrate the importance of pontine GABAergic pathways in shaping the response to hypoxia.     

Apneusis follows disruption of NMDA-type glutamate receptors in vagotomized ground squirrels

The influences of N-methyl-D-aspartate (NMDA) type glutamate receptor antagonism, by (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]-cyclohepten-5,10-imine maleate (MK-801), on breathing pattern and ventilatory chemoresponses, were assessed in anaesthetized vagotomized spontaneously breathing golden-mantled ground squirrels, Spermophilus lateralis. MK-801 was administered by either bilateral pressure micro-injection into a region of the rostral dorsolateral pons, containing the medial and lateral Parabrachial and K?lliker-Fuse nuclei (the Parabrachial complex, PbC), or by systemic injection. Both treatments induced apneusis. These data indicate that functional NMDA receptor-mediated processes located within the PbC terminate inspiration and actively prevent apneusis in vagotomized ground squirrels. Although both hypercapnia and hypoxia stimulated breathing during the apneusis, the responses were generally slight. The breathing frequency component of the hypercapnic ventilatory response was completely eliminated supporting the hypothesis that the PbC is an integral component of the control network for CO(2) chemoreflex responses. Differences in the results of systemic versus PbC MK-801 illustrate that NMDA receptor-mediated processes outside the PbC do influence ventilation. Our data also show that such processes outside the PbC lengthen both inspiration and expiration in this species, slowing ventilation, and are necessary for the expression of the hypoxic ventilatory response.     

Mapping of CGRP in the alpaca (Lama pacos) brainstem

In this study, we demonstrate the presence of immunoreactive structures containing calcitonin gene-related peptide in the alpaca brainstem. This is the first time that a detailed mapping of the cell bodies and fibers containing this neuropeptide in the alpaca brainstem has been carried out using an immunocytochemical technique. Immunoreactive cell bodies and fibers were widely distributed throughout the alpaca brainstem. A high density of calcitonin gene-related peptide-immunoreactive perikarya was found in the superior colliculus, the dorsal nucleus of the raphe, the trochlear nucleus, the lateral division of the marginal nucleus of the brachium conjunctivum, the motor trigeminal nucleus, the facial nucleus, the pons reticular formation, the retrofacial nucleus, the rostral hypoglossal nucleus, and in the motor dorsal nucleus of the vagus, whereas a high density of fibers containing calcitonin gene-related peptide was observed in the lateral division of the marginal nucleus of the brachium conjunctivum, the parvocellular division of the alaminar spinal trigeminal nucleus, the external cuneate nucleus, the nucleus of the solitary tract, the laminar spinal trigeminal nucleus, and in the area postrema. This widespread distribution indicates that the neuropeptide studied might be involved in multiple functions in the alpaca brainstem.     

Reversal of noradrenergic depletion and lipid peroxidation in the pons after brain injury correlates with motor function recovery in rats

Functional impairment after brain injury (BI) has been attributed to the inhibition of regions that are related to the injured site. Therefore, noradrenaline (NA) is thought to play a critical role in recovery from motor injury. However, the mechanism of this recovery process has not been completely elucidated. Moreover, the locus coeruleus (LC) projects from the pons through the rat sensorimotor cortex, and injury axotomizes LC fibers, depressing NA function. This was tested by measuring lipid peroxidation (LP) in the pons after sensorimotor cortex injury. Depression of function in the pons would be expected to alter areas receiving pontine efferents. Male Wistar rats were divided into three groups: control (n=16), injured (n=10) and recovering (n=16), and they were evaluated using a beam-walking assay between 2 and 20 days after cortical injury. We performed measures of NA and LP in both sides of the pons and cerebellum. We found a decrease of NA in the pons and the cerebellum, and a concomitant increase in the motor deficit and LP in the pons of injured animals. Recovering rats had NA and LP levels that were very similar to those observed in control rats. These observations suggest that the mechanism of remote inhibition after BI involves lipid peroxidation, and that the NA decrease found in the cerebellum of injured animals is mediated by a noradrenergic depression in the pons, or in areas receiving NA projections from the pons.     

Locus coeruleus noradrenergic neurons and CO2 drive to breathing

The Locus coeruleus (LC) has been suggested as a CO₂ chemoreceptor site in mammals. In the present study, we assessed the role of LC noradrenergic neurons in the cardiorespiratory and thermal responses to hypercapnia. To selectively destroy LC noradrenergic neurons, we administered 6-hydroxydopamine (6-OHDA) bilaterally into the LC of male Wistar rats. Control animals had vehicle (ascorbic acid) injected (sham group) into the LC. Pulmonary ventilation (plethysmograph), mean arterial pressure (MAP), heart rate (HR), and body core temperature (T _c, data loggers) were measured followed by 60 min of hypercapnic exposure (7% CO₂ in air). To verify the correct placement and effectiveness of the chemical lesions, tyrosine hydroxylase immunoreactivity was performed. Hypercapnia caused an increase in pulmonary ventilation in all groups, which resulted from increases in respiratory frequency and tidal volume (V _T) in sham-operated and 6-OHDA-lesioned groups. The hypercapnic ventilatory response was significantly decreased in 6-OHDA-lesioned rats compared with sham group. This difference was due to a decreased V _T in 6-OHDA rats. LC chemical lesion or hypercapnia did not affect MAP, HR, and T _c. Thus, we conclude that LC noradrenergic neurons modulate hypercapnic ventilatory response but play no role in cardiovascular and thermal regulation under resting conditions.