Hypoglossal Rootlets Research Articles

Oxidative stress in retrotrapezoid nucleus/parafacial respiratory group and impairment of central chemoreception in rat offspring exposed to maternal cigarette smoke

We have reported that smoking during pregnancy is associated with deficit in neonatal central chemoreception. However, the underlying mechanism is not well clarified. In this study, we developed a rat model of maternal cigarette smoke (CS) exposure. Pregnant rats were exposed to CS during gestational day 1−20. Offspring were studied on postnatal day 2. Reactive oxygen species (ROS) content and expressions of antioxidant proteins in retrotrapezoid nucleus/parafacial respiratory group (RTN/pFRG) were examined by fluorogenic dye MitoSOX™ Red and Western blotting, respectively. The response of hypoglossal rootlets discharge to acidification was also detected with micro-injection of H2O2 into RTN/pFRG of offspring brainstem slices in vitro. Results showed that maternal CS exposure led to an increase in ROS production, and brought about decreases in mitochondrial superoxide dismutase and Kelch-like ECH-associated protein-1, and an increase in NF-E2-related factor 2 in offspring RTN/pFRG. Catalase and glutathione reductase expressions were not significantly changed. Moreover, oxidative stress induced by micro-injection of H2O2 into RTN/pFRG in vitro inhibited the discharge response of hypoglossal rootlets to acidification. These findings suggest that maternal CS exposure results in oxidative stress in RTN/pFRG of rat offspring, which might play a role in the impairment of central chemoreception.

Far Lateral Approach for Clipping of a Posterior Inferior Cerebellar Artery Aneurysm.

Objectives To describe a far lateral approach for microsurgical clipping of a ruptured posterior inferior cerebellar artery (PICA) aneurysm involving the hypoglossal nerve, with emphasis on the microsurgical anatomy, and technique. Design A far lateral craniotomy is performed in the lateral decubitus position and the transverse and sigmoid sinuses were exposed. After opening the dura, sutures are placed to allow gentle mobilization of the sinuses. The ipsilateral cerebellar tonsil is mobilized and the PICA is followed to its junction with the vertebral artery. Hypoglossal nerve rootlets are draped over the dome of the aneurysm. Mobilization of the PICA and the hypoglossal nerve away from the lateral medulla allows microsurgical clipping of the aneurysm neck. Photographs of the region are borrowed from Dr Rhoton's laboratory to illustrate the microsurgical anatomy. Participants The senior authors performed the surgery. The video was edited by Drs. V.N. and J.B. Chart review and literature review were performed by Drs. W.M. and J.B. Outcome Measures Outcome was assessed with successful clip occlusion and postoperative neurological function. Results There was complete clip occlusion of the PICA aneurysm with no postoperative neurological deficits. The patient was discharged home after an uneventful hospital course. Conclusion The far lateral approach provides an adequate corridor to the ventrolateral brainstem for microsurgical treatment of PICA aneurysms. An adequate understanding of the relevant microsurgical anatomy is the key to safe and effective clipping in this region. The link to the video can be found at: https://youtu.be/yhjKRIG5H74 .

Anatomical Analysis of the Vagoaccessory Triangle and the Triangles Within: The Suprahypoglossal, Infrahypoglossal, and Hypoglossal–Hypoglossal Triangles

Rootlets of the lower cranial nerves create a web that limits microsurgical maneuverability in the cerebellomedullary cistern (CMC). The vagoaccessory triangle (VAT) and 2 triangles within it were defined to identify surgical routes to aneurysms of the posterior inferior cerebellar artery (PICA). Dividing the VAT into 2 triangles (suprahypoglossal [SHT] and infrahypoglossal [IHT]), although elegant, oversimplifies CMC anatomy. The triangle formed by the superior and inferior hypoglossal rootlets (hypoglossal-hypoglossal triangle [HHT]) needs consideration as well. Far-lateral craniotomy was performed on 10 cadaveric heads bilaterally. Areas of CMC triangles were calculated. Relationships between the PICA origin and the adjacent triangles were analyzed. Vertebral artery (VA) exposure and clipping lengths were recorded for each triangle. The area of SHT was almost twice those of IHT and HHT. The average VA depth relative to VAT increased significantly when moving distally from proximal VA (P < 0.001), but VA exposure and clipping lengths were not significantly different between triangles. IHT, HHT, and SHT defined VA subsegments (V4i, V4h, and V4s), with PICA most commonly originating from V4h. Based on our cadaveric measurements, the V4 subsegments were identified with 76% accuracy in angiograms. Based on this study, VAT should be divided into 3 triangles, not 2. Splayed rootlets of cranial nerve XII and multiple outlet foramina create an important space different from the previously recognized SHT and IHT. These triangles provide corridors to vascular pathologies. V4 subsegments may be approximated from imaging studies and may help with preoperative planning.

The Endoscopic Endonasal Approach to the Hypoglossal Canal: The Role of the Eustachian Tube as a Landmark for Dissection.

Improvements in endoscopic technology and reconstructive techniques have made the endoscopic endonasal approach (EEA) a viable option to approach ventromedial lesions in the region of the hypoglossal canal. Prior to contemplating this surgical corridor, a thorough understanding of anatomic relationships and landmarks is essential to safely approach this region of the posterior skull base through an EEA. To describe the surgical technique and anatomic landmarks in the EEA to the hypoglossal canal through referencing nasopharyngeal and posterior skull base anatomy. Study of latex-injected cadaveric heads at the North Carolina Eye Bank Multidisciplinary Surgical Skills Laboratory at the University of North Carolina. An EEA to the hypoglossal canal was carried out bilaterally in 5 embalmed, latex-injected cadaver heads. Cadaveric measurements of anatomic landmarks and relationships in the approach were obtained using a 10-cm surgical ruler and were reported as mean distances. Additionally, high-quality endoscopic images demonstrating the operative technique and anatomic relationships were obtained. The distance between the lacerum segment of the internal carotid arteries, the superolateral boundary, was 23.6 mm (SD, 11.8 mm). The distance between the anterolateral edge of the occipital condyles, the inferolateral boundary, was 19 mm (SD, 0.80 mm). The supracondylar groove was identified in the same anteroposterior plane as the nasopharyngeal orifice of the eustachian tube, and the anterior-most edge of the occipital condyle was 14 mm (SD, 0.82 mm) from the posterosuperior edge of the salpingopharyngeal fold. Additionally, the transtubercular corridor was on the same plane as the superior edge of the torus tubarius in the anteroposterior axis. The distance to the hypoglossal canal from midline was 10 mm, which was found after completing drilling in the transcondylar and transtubercular corridors. Last, the hypoglossal nerve rootlets were identified entering the canal 6 mm inferiorly and 8 mm laterally from the vertebrobasilar junction. The eustachian tube and other elements of nasopharyngeal anatomy are fixed landmarks that provide important points of reference when approaching the hypoglossal canal through an EEA. A thorough understanding of these anatomic relationships is vital in safely navigating this direct, surgical corridor to the posterior fossa.

Impairment of Central Chemoreception in Neonatal Rats Induced by Maternal Cigarette Smoke Exposure during Pregnancy.

It has been postulated that prenatal cigarette smoke exposure (CSE) increases the risk for sudden infant death syndrome. The victims of infant death syndrome suffer from respiratory abnormalities, such as central apnea, diminished chemoreflex and alteration in respiratory pattern during sleep. However, no experimental evidence on CSE model exists to confirm whether prenatal CSE gives rise to reduction of neonatal central chemoreception in in vitro preparations in absence of peripheral sensory feedback. The aim of the present study was to test the hypothesis that maternal CSE during pregnancy depresses central chemoreception of the neonatal rats. The pregnant rats were divided into two groups, control (n = 8) and CSE (n = 8). Experiments were performed on neonatal (0–3days) rat pups. Fictive respiratory activity was monitored by recording the rhythmic discharge from the hypoglossal rootlets of the medullary slices obtained from the neonatal rats. The burst frequency (BF) and integrated amplitude (IA) of the discharge were analyzed. Their responses to acidified artificial cerebrospinal fluid (aCSF) were tested to indicate the change of the central chemosensitivity. Under condition of perfusing with standard aCSF (pH 7.4), no significant difference was detected between the two groups in either BF or IA (P>0.05). Under condition of perfusing with acidified aCSF (pH 7.0), BF was increased and IA was decreased in both groups (P<0.01). However, their change rates in the CSE group were obviously smaller than that in the control group, 66.98 ± 10.11% vs. 143.75 ± 15.41% for BF and −22.38 ± 2.51% vs. −44.90 ± 3.92% for IA (P<0.01). In conclusion, these observations, in a prenatal CSE model, provide important evidence that maternal smoking during pregnancy exerts adverse effects on central chemoreception of neonates.

KATP channels of parafacial respiratory group (pFRG) neurons are involved in H2S-mediated central inhibition of respiratory rhythm in medullary slices of neonatal rats

We recently found that hydrogen sulfide (H2S) participates in inhibitory regulation of rhythmic respiration by acting on the parafacial respiratory group (pFRG) in medullary slices of neonatal rats. The present study investigated whether ATP-sensitive potassium (KATP) channels are expressed in neurons of the pFRG, and, if so, whether they play a role in central regulation of respiratory activity, in particular the H2S-mediated central inhibition of respiratory rhythm in medullary slices of neonatal rats. Immunohistochemical techniques revealed that KATP channels are expressed in neurons of the pFRG region. Micro-injection of the KATP channel activator, pinacidil, into the pFRG region inhibited the discharge rhythm of hypoglossal rootlets, whereas injection of the KATP channel blocker, glibenclamide (Gl), had no effect. Micro-injection of the H2S donor sodium hydrosulfide (NaHS) into the pFRG region produced identical inhibitory responses to those induced by pinacidil. However, combined micro-injection of Gl and NaHS eliminated inhibitory effects of NaHS and converted to minor excitatory effects on the respiratory rhythm. It can be concluded that KATP channels of pFRG neurons are involved in the central regulation of respiratory rhythm and H2S-mediated inhibitory actions on respiratory rhythm in medullary slices of neonatal rats.

The anti-apoptotic effect of hydrogen sulfide attenuates injuries to the medullary respiratory centers of neonatal rats subjected to in utero cigarette smoke exposure

H2S may serve as an important neuroprotectant. The present experiments were performed to determine whether H2S could attenuate the injuries sustained by the medullary respiratory centers of neonatal rats that were subjected to cigarette smoke exposure (CS) in utero. Pregnant SD rats were divided into 4 exposure groups: control, CS, CS+NaHS (donor of H2S) and NaHS. Hypoxia decreased the burst frequencies of the hypoglossal rootlets of the medullary slices in CS neonatal rats, and NaHS offset the hypoxia-induced respiratory suppression. Nissl staining indicated that NaHS alleviated the injuries that were sustained by neurons after CS in utero. NaHS also decreased the number of TUNEL-positive neurons and the expression of activated caspase-3 protein in the medulla oblongata of CS neonatal rats. Furthermore, NaHS promoted Bcl-2 protein expression and reduced Bax protein and mRNA expression in the medulla oblongata of CS neonatal rats. Therefore, the present study indicates that the anti-apoptotic effect of H2S protects rat medullary respiratory centers from injuries that would otherwise be sustained from in utero CS exposure.

Perinatal sulfur dioxide exposure alters brainstem parasympathetic control of heart rate.

Sulfur dioxide (SO₂) is an air pollutant that impedes neonatal development and induces adverse cardiorespiratory health effects, including tachycardia. Here, an animal model was developed that enabled characterization of (i) in vivo alterations in heart rate and (ii) altered activity in brainstem neurons that control heart rate after perinatal SO₂ exposure. Pregnant Sprague-Dawley dams and their pups were exposed to 5 parts per million SO₂ for 1 h daily throughout gestation and 6 days postnatal. Electrocardiograms were recorded from pups at 5 days postnatal to examine changes in basal and diving reflex-evoked changes in heart rate following perinatal SO₂ exposure. In vitro studies employed whole-cell patch-clamp electrophysiology to examine changes in neurotransmission to cardiac vagal neurons within the nucleus ambiguus upon SO₂ exposure using a preparation that maintains fictive inspiratory activity recorded from the hypoglossal rootlet. Perinatal SO₂ exposure increased heart rate and blunted the parasympathetic-mediated diving reflex-evoked changes in heart rate. Neither spontaneous nor inspiratory-related inhibitory GABAergic or glycinergic neurotransmission to cardiac vagal neurons was altered by SO₂ exposure. However, excitatory glutamatergic neurotransmission was decreased by 51.2% upon SO₂ exposure. This diminished excitatory neurotransmission was tetrodotoxin-sensitive, indicating SO₂ exposure impaired the activity of preceding glutamatergic neurons that synapse upon cardiac vagal neurons. Diminished glutamatergic, but unaltered inhibitory neurotransmission to cardiac vagal neurons provides a mechanism for the observed SO₂-induced elevated heart rate via an impairment of brainstem cardioinhibitory parasympathetic activity to the heart.

Site-specific hydrogen sulfide-mediated central regulation of respiratory rhythm in medullary slices of neonatal rats

Hydrogen sulfide (H2S) is involved in central regulation of respiratory rhythm at the level of the medulla oblongata. The present study was carried out to test our hypothesis that H2S exerts site-specific regulatory action on respiratory rhythm in the medulla oblongata of neonatal rats. The rhythmic discharge of hypoglossal rootlets in medullary slices of neonatal rats was recorded. 200μM NaHS (an H2S donor) increased burst frequency (BF) in 900-μm slices containing the pre-Bötzinger complex (preBötC), whereas it caused diphasic responses in 1200-, 1400- and 1800-μm slices containing both the preBötC and part or all of the parafacial respiratory group (pFRG): an initial decrease in BF followed by an increase. The initial decrease in BF was no longer observed after unilateral lesion of the pFRG region in the 1400-μm slices. In addition, BF was increased by a unilateral micro-injection of NaHS into the preBötC region, but was decreased by an injection into the pFRG region. These data support our hypothesis that the regulatory action of H2S on respiratory rhythm in the medulla oblongata is site-specific. The excitatory effect is caused by the preBötC, while the inhibitory effect is from the pFRG.

Modulation of Bulbospinal Rostral Ventral Lateral Medulla Neurons by Hypoxia/Hypercapnia but Not Medullary Respiratory Activity

Although sympathetic vasomotor discharge has respiratory modulation, the site(s) responsible for this cardiorespiratory interaction is unknown. One likely source for this coupling is the rostral ventral lateral medulla (RVLM), where presympathetic neurons originate in close apposition to respiratory neurons. The current study tested the hypothesis that RVLM bulbospinal neurons are modulated by medullary respiratory network activity using whole-cell patch-clamp electrophysiological recordings of RVLM neurons while simultaneously recording fictive respiratory bursting activity from the hypoglossal rootlet. Additionally, we examined whether challenges to cardiorespiratory function, mainly hypoxia/hypercapnia, alter the activity of bulbospinal neurons and, secondarily, whether changes in synaptic input mediate these responses. Surprisingly, our results indicate that inspiratory-related activity did not modulate glutamatergic, γ-aminobutyric acid-ergic, or glycinergic synaptic events or spontaneous action potential firing in these RVLM neurons. However, hypoxia/hypercapnia reversibly decreased the frequency of γ-aminobutyric acid and glycine inhibitory postsynaptic currents. Glycinergic inhibitory postsynaptic current frequency was depressed from the fifth through the 10th minute, whereas the depression of γ-aminobutyric acid-ergic events became significant only at the 10th minute of hypoxia/hypercapnia. On the basis of spontaneous firing activity, there were 2 populations of RVLM bulbospinal neurons. The firing frequency of low-discharging RVLM neurons was facilitated by hypoxia/hypercapnia, and this increase depended on reduced inhibitory neurotransmission. The firing frequency in RVLM neurons with high-discharge rates was inhibited, independent of synaptic input, by hypoxia/hypercapnia. This article demonstrates that sympathetic-respiratory coupling is not active in the neonatal brain stem slice, and reductions in inhibitory neurotransmission to low spontaneously active bulbospinal RVLM neurons are responsible for hypoxia/hypercapnia-elicited increases in activity.

Contribution of BKCa channels of neurons in rostral ventrolateral medulla to CO-mediated central regulation of respiratory rhythm in medullary slices of neonatal rats

We recently described that carbon monoxide (CO) participated in the regulation of rhythmic respiration in medullary slices. The present study was undertaken to further assess whether the large-conductance calcium-activated potassium channels (BKCa channels) are involved in the CO-mediated central regulation of respiratory rhythm in medullary slices. The rhythmic discharge of hypoglossal rootlets of medullary slices of neonatal rats was recorded. We observed that blocking BKCa channels could partially abolish the effects of CO on the rhythmic bursts of hypoglossal rootlets. With whole-cell patch-clamp recording technique, we further observed that CO could reversibly augment potassium current density of the neurons in the rostral ventrolateral medulla. The CO-induced increase in potassium current was entirely blocked by the pretreatment of slices with BKCa channels blocker; whereas blockade of CO generation with zinc protoporphyrin-IX produced an opposite response. Altogether, these data indicate that BKCa channels of the neurons in neonatal rostral ventrolateral medulla could be activated by CO and involved in CO-mediated central regulation of respiratory rhythm in medullary slices.

Developmental changes in NMDA receptor subunits alter burst frequency of neonatal rat brainstem slices containing the preBötzinger Complex

N‐methyl‐d‐aspartate receptors (NMDARs) are crucial in controlling synaptic excitation and plasticity at glutamatergic synapses in the CNS. NMDARs are composed of multiple NR1 and at least one of four NR2 subunits and previous studies have shown development changes in NMDARs. In the perinatal brain, NR2B subunits predominate, however these subunits are supplemented with NR2A/NR2C subunits. We hypothesized that respiratory rhythm in the brainstem of postnatal rats (P0–P3), is developmentally modulated by NMDAR subunit changes in the preBötzinger Complex (pBC). We prepared pBC‐containing slices from Sprague Dawley rat pups (P0 to P5) and measured changes in hypoglossal (XII) rootlet output and characterized the switch from NR2B‐ to NR2A‐containing NMDARs by bath application of 10 μM ifenprodil, a noncompetitive inhibitor of the NR2B subunit. Our results show a significant decrease (p &lt; 0.05; compared to baseline) in inter‐burst interval after application of ifenprodil in P0–P3 (n=3) slices with no significant change in animals &gt; P3. This suggests that NR2B‐containing NMDARs play a role in the modulation of respiratory rhythm early in life. Support: This work was supported by NIH/NHLBI grant HL081622

Nitric Oxide and ATP-Sensitive Potassium Channels Mediate Lipopolysaccharide-Induced Depression of Central Respiratory-Like Activity in Brain Slices

Infection may result in early abnormalities in respiratory movement, and the mechanism may involve central and peripheral factors. Peripheral mechanisms include lung injury and alterations in electrolytes and body temperature, but the central mechanisms remain unclear. In the present study, brainstem slices harvested from rats were stimulated with lipopolysaccharide at different doses. Central respiratory activities as demonstrated by electrophysiological activity of the hypoglossal rootlets were examined and the mechanisms were investigated by inhibiting nitric oxide synthase and ATP-sensitive potassium channels. As a result, 0.5 µg/ml lipopolysaccharide mainly caused inhibitory responses in both the frequency and the output intensity, while 5 µg/ml lipopolysaccharide caused an early frequency increase followed by delayed decreases in both the frequency and the output intensity. At both concentrations the inhibitory responses were fully reversed by inhibition of nitric oxide synthase with Nω-nitro-L-arginine methyl ester hydrochloride (20 µM), and by inhibition of ATP- sensitive potassium channels with glybenclamide (100 µM). These results show that direct lipopolysaccharide challenge altered central respiratory activity in dose- and time- related manners. Nitric oxide synthase and ATP-sensitive potassium channels may be involved in the respiratory changes.

Cranial nerve fasciculation and Schwann cell migration are impaired after loss of Npn-1

Interaction of the axon guidance receptor Neuropilin-1 (Npn-1) with its repulsive ligand Semaphorin 3A (Sema3A) is crucial for guidance decisions, fasciculation, timing of growth and axon–axon interactions of sensory and motor projections in the embryonic limb. At cranial levels, Npn-1 is expressed in motor neurons and sensory ganglia and loss of Sema3A–Npn-1 signaling leads to defasciculation of the superficial projections to the head and neck. The molecular mechanisms that govern the initial fasciculation and growth of the purely motor projections of the hypoglossal and abducens nerves in general, and the role of Npn-1 during these events in particular are, however, not well understood. We show here that selective removal of Npn-1 from somatic motor neurons impairs initial fasciculation and assembly of hypoglossal rootlets and leads to reduced numbers of abducens and hypoglossal fibers. Ablation of Npn-1 specifically from cranial neural crest and placodally derived sensory tissues recapitulates the distal defasciculation of mixed sensory-motor nerves of trigeminal, facial, glossopharyngeal and vagal projections, which was observed in Npn-1−/− and Npn-1Sema− mutants. Surprisingly, the assembly and fasciculation of the purely motor hypoglossal nerve are also impaired and the number of Schwann cells migrating along the defasciculated axonal projections is reduced. These findings are corroborated by partial genetic elimination of cranial neural crest and embryonic placodes, where loss of Schwann cell precursors leads to aberrant growth patterns of the hypoglossal nerve. Interestingly, rostral turning of hypoglossal axons is not perturbed in any of the investigated genotypes. Thus, initial hypoglossal nerve assembly and fasciculation, but not later guidance decisions depend on Npn-1 expression and axon–Schwann cell interactions.

Protective action of endogenously generated H2S on hypoxia-induced respiratory suppression and its relation to antioxidation and down-regulation of c-fos mRNA in medullary slices of neonatal rats

We previously reported that exogenous H(2)S played roles in protection of respiratory centers against hypoxic injury in medullary slices of neonatal rats. The protective action of endogenous H(2)S and its relation to antioxidation and down-regulation of c-fos mRNA were investigated in the present study. Perfusion of the slices with l-cysteine (Cys), substrate of cystathionine β-synthase (CBS, H(2)S synthase), could increase frequency of rhythmic respiratory discharge of the hypoglossal rootlets and prevent respiratory suppression induced by hypoxia, whereas perfusion with hydroxylamine (NH(2)OH, inhibitor of CBS) could postpone recovery of respiration from hypoxic inhibition. NH(2)OH also significantly enhanced hypoxia-induced increase in malondialdehyde (MDA) content of the slices. The hypoxia-induced up-regulation of c-fos mRNA could be markedly antagonized by S-adenosyl-l-methionine (SAM, activator of CBS), but greatly increased by NH(2)OH. Neither NH(2)OH, Cys nor SAM had any effect on expression of bcl-2 mRNA in hypoxic medullary slices. These results indicate that endogenously generated H(2)S was involved in protection of the medullary respiratory centers against hypoxic injury partly via antioxidation and down-regulation of c-fos.

Population calcium imaging of spontaneous respiratory and novel motor activity in the facial nucleus and ventral brainstem in newborn mice

The brainstem contains rhythm and pattern forming circuits, which drive cranial and spinal motor pools to produce respiratory and other motor patterns. Here we used calcium imaging combined with nerve recordings in newborn mice to reveal spontaneous population activity in the ventral brainstem and in the facial nucleus. In Fluo-8AM loaded brainstem-spinal cord preparations, respiratory activity on cervical nerves was synchronized with calcium signals at the ventrolateral brainstem surface. Individual ventrolateral neurons at the level of the parafacial respiratory group showed perfect or partial synchrony with respiratory nerve bursts. In brainstem-spinal cord preparations, cut at the level of the mid-facial nucleus, calcium signals were recorded in the dorsal, lateral and medial facial subnuclei during respiratory activity. Strong activity initiated in the dorsal subnucleus, followed by activity in lateral and medial subnuclei. Whole-cell recordings from facial motoneurons showed weak respiratory drives, and electrical field potential recordings confirmed respiratory drive to particularly the dorsal and lateral subnuclei. Putative facial premotoneurons showed respiratory-related calcium signals, and were predominantly located dorsomedial to the facial nucleus. A novel motor activity on facial, cervical and thoracic nerves was synchronized with calcium signals at the ventromedial brainstem extending from the level of the facial nucleus to the medulla–spinal cord border. Cervical dorsal root stimulation induced similar ventromedial activity. The medial facial subnucleus showed calcium signals synchronized with this novel motor activity on cervical nerves, and cervical dorsal root stimulation induced similar medial facial subnucleus activity. In conclusion, the dorsal and lateral facial subnuclei are strongly respiratory-modulated, and the brainstem contains a novel pattern forming circuit that drives the medial facial subnucleus and cervical motor pools.

Nitric oxide synthase expression in the medullary respiratory related nuclei and its involvement in CO-mediated central respiratory effects in neonatal rats

The present study was conducted in order to observe the potential participation of the nitric oxide synthase–NO pathway in CO-mediated regulation of respiration of neonatal rats. An immunofluorescent histochemical technique was used to examine the existence of the neuronal nitric oxide synthase, a key enzyme of synthesizing NO, in medullary respiratory nuclei. The rhythmic respiratory-like discharges of hypoglossal rootlets of medullary slices were recorded to test the role of the nitric oxide synthase in CO-mediated respiratory effects. We observed neuronal nitric oxide synthase expressed in the medullary respiratory nuclei in conjunction with CO lengthened expiratory duration, decreased respiratory frequency, and increased inspiratory amplitude. These CO-mediated respiratory effects could be partially eliminated by prior treatment of the slices with Nω-nitro- l-arginine methyl ester, an inhibitor of nitric oxide synthase. The results suggest that nitric oxide synthase–NO pathway might be involved in the CO-mediated central regulation of respiration at the level of medulla oblongata in neonatal rats.

Resolution of Isolated Unilateral Hypoglossal Nerve Palsy Following Microvascular Decompression of the Intracranial Vertebral Artery

Isolated hypoglossal nerve paresis due to mechanical compression from a vascular lesion is very rare. We present a case of a 32-year-old man who presented with spontaneous abrupt-onset dysarthria, swallowing difficulty and left-sided tongue atrophy. Brain computed tomographic angiography and magnetic resonance imaging of the brainstem demonstrated an abnormal course of the left vertebral artery compressing the medulla oblongata at the exit zone of the hypoglossal rootlets that was relieved by microvascular decompression of the offending intracranial vertebral artery. This case supports the hypothesis that hypoglossal nerve palsy can be due to nerve stretching and compression by a pulsating normal vertebral artery. Microvascular decompression of the intracranial nerve and careful evaluation of the imaging studies can resolve unexpected isolated hypoglossal nerve palsy.

Large-conductance calcium-activated potassium channels in the neurons of pre-Bötzinger complex and their participation in the regulation of central respiratory activity in neonatal rats

The present study was conducted to test our hypothesis that the large-conductance calcium-activated potassium channels (BK Ca channels) exist in the neurons of the pre-Bötzinger complex (PBC), a brainstem region that may generate respiratory rhythm in mammals, and play roles in central regulation of respiratory activity in neonatal rats. Immunohistochemical technique revealed that BK Ca channels expressed in the neurons of PBC region. Whole cell voltage clamp recordings from the neurons in the PBC showed that BK Ca channels could be activated by membrane depolarization and blocked by 1 mM tetraethylammonium (TEA) or 10 μM paxilline in the preparation of thin (about 300 μm) medullary slices of neonatal rats. The rhythmic respiratory-like discharge of hypoglossal rootlets could be changed by perfusing the thick (700–900 μm) medullary slices with 1 mM TEA or 10 μM paxilline. Both TEA and paxilline could prolong the inspiratory duration, shorten the expiratory duration and increase the respiratory frequency. The results suggest that BK Ca channels exist in the PBC neurons and may be involved in the central control of rhythmic respiration in the neonatal rats.

Protective effect of hydrogen sulfide on hypoxic respiratory suppression in medullary slice of neonatal rats

Hydrogen sulfide (H(2)S) has been shown to play a protective role in injury of cells induced by hypoxia. Little is known however about its effect on medullary hypoxia-induced rhythmic respiratory suppression. In the present study, a decrease in frequency of rhythmic discharge of hypoglossal rootlets was observed in medullary slices of neonatal rats perfused with 95% N(2)-5% CO(2) to produce hypoxia. Perfusion with NaHS (H(2)S donor) prevented the inhibitory effect of hypoxia on the burst activity of the rootlets, whereas such action of NaHS was suppressed by pretreatment with glibenclamide, a blocker of K(ATP) channels. In addition, the increase in malondialdehyde content and the up-regulation of c-fos mRNA expression of the slices induced by hypoxia was significantly reduced by NaHS. These results indicate that exogenous H(2)S may protect the medullary respiratory center against hypoxic injury via activation of K(ATP) channels, reduction of lipid peroxidation and down-regulation of c-fos.