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      Coordinated Respiratory Motor Activity in Nerves Innervating the Upper Airway Muscles in Rats

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          Abstract

          Maintaining the patency of the upper airway during breathing is of vital importance. The activity of various muscles is related to the patency of the upper airway. In the present study, we examined the respiratory motor activity in the efferent nerves innervating the upper airway muscles to determine the movements of the upper airway during respiration under normocapnic conditions (pH = 7.4) and in hypercapnic acidosis (pH = 7.2). Experiments were performed on arterially perfused decerebrate rats aged between postnatal days 21–35. We recorded the efferent nerve activity in a branch of the cervical spinal nerve innervating the infrahyoid muscles (CN), the hypoglossal nerve (HGN), the external branch of the superior laryngeal nerve (SLN), and the recurrent laryngeal nerve (RLN) with the phrenic nerve (PN). Inspiratory nerve discharges were observed in all these nerves under normocapnic conditions. The onset of inspiratory discharges in the CN and HGN was slightly prior to those in the SLN and RLN. When the CO 2 concentration in the perfusate was increased from 5% to 8% to prepare for hypercapnic acidosis, the peak amplitudes of the inspiratory discharges in all the recorded nerves were increased. Moreover, hypercapnic acidosis induced pre-inspiratory discharges in the CN, HGN, SLN, and RLN. The onset of pre-inspiratory discharges in the CN, HGN, and SLN was prior to that of discharges in the RLN. These results suggest that the securing of the airway that occurs a certain time before dilation of the glottis may facilitate ventilation and improve hypercapnic acidosis.

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          A working heart-brainstem preparation of the mouse.

          An intra-arterially perfused working heart-brainstem preparation (WH-BP) was developed to allow studies into functionally identified cardiovascular and respiratory neurones in an in vitro milieu. This report provides that first description of this preparation. Evidence is presented indicating that the WH-BP: (1) spontaneously generates eupneic-like phrenic nerve activity, indicative of adequate oxygenation of the brainstem; (2) preserves the integrity of central coupling between the central respiratory rhythm generator and cardiac vagal motor neurones and, (3), allow an intracellular analysis of medullary cardio-respiratory neurones. The WH-BP may provide an advantaged environment for analysis of both synaptic and cellular mechanisms within the medulla that regulate cardio-respiratory activity.
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            Role of inhibition in respiratory pattern generation.

            Postsynaptic inhibition is a key element of neural circuits underlying behavior, with 20-50% of all mammalian (nongranule) neurons considered inhibitory. For rhythmic movements in mammals, e.g., walking, swimming, suckling, chewing, and breathing, inhibition is often hypothesized to play an essential rhythmogenic role. Here we study the role of fast synaptic inhibitory neurotransmission in the generation of breathing pattern by blocking GABA(A) and glycine receptors in the preBötzinger complex (preBötC), a site essential for generation of normal breathing pattern, and in the neighboring Bötzinger complex (BötC). The breathing rhythm continued following this blockade, but the lung inflation-induced Breuer-Hering inspiratory inhibitory reflex was suppressed. The antagonists were efficacious, as this blockade abolished the profound effects of the exogenously applied GABA(A) receptor agonist muscimol or glycine, either of which under control conditions stopped breathing in vagus-intact or vagotomized, anesthetized, spontaneously breathing adult rats. In vagotomized rats, GABA(A)ergic and glycinergic antagonists had little, if any, effect on rhythm. The effect in vagus-intact rats was to slow the rhythm to a pace equivalent to that seen after suppression of the aforementioned Breuer-Hering inflation reflex. We conclude that postsynaptic inhibition within the preBötC and BötC is not essential for generation of normal respiratory rhythm in intact mammals. We suggest the primary role of inhibition is in shaping the pattern of respiratory motor output, assuring its stability, and in mediating reflex or volitional apnea, but not in the generation of rhythm per se.
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              Respiratory rhythm generation during gasping depends on persistent sodium current.

              In severe hypoxia, homeostatic mechanisms maintain function of the brainstem respiratory network. We hypothesized that hypoxia involves a transition from neuronal mechanisms of normal breathing (eupnea) to a rudimentary pattern of inspiratory movements (gasping). We provide evidence for hypoxia-driven transformation within the central respiratory oscillator, in which gasping relies on persistent sodium current, whereas eupnea does not depend on this cellular mechanism.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                10 November 2016
                2016
                : 11
                : 11
                : e0166436
                Affiliations
                [1 ]Department of Oral Physiology, Showa University School of Dentistry, Shinagawa-ku, Tokyo 142–8555, Japan
                [2 ]Department of Perioperative Medicine, Division of Anesthesiology, Showa University School of Dentistry, Oota-ku, Tokyo 145–8515, Japan
                Texas Christian University, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: KN T. Inoue.

                • Data curation: ST KN SN.

                • Formal analysis: ST KN AM.

                • Funding acquisition: KN T. Inoue.

                • Investigation: ST.

                • Methodology: ST KN.

                • Project administration: KN T. Inoue.

                • Supervision: T. Iijima.

                • Validation: SN AM.

                • Writing – original draft: ST KN.

                • Writing – review & editing: KN T. Inoue.

                Article
                PONE-D-16-06619
                10.1371/journal.pone.0166436
                5104329
                27832132
                f9a970ca-1978-4927-821b-86ab16442e2f
                © 2016 Tachikawa et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 16 February 2016
                : 28 October 2016
                Page count
                Figures: 5, Tables: 2, Pages: 14
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100001691, Japan Society for the Promotion of Science;
                Award ID: 25350622
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001691, Japan Society for the Promotion of Science;
                Award ID: 26293397
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001691, Japan Society for the Promotion of Science;
                Award ID: 15K15687
                Award Recipient :
                This work was supported in part by MEXT/ JSPS KAKENHI ( www.jsps.go.jp) Grant number 25350622 (KN), 26293397 (T. Inoue), and 15K15687 (T. Inoue). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Medicine and Health Sciences
                Diagnostic Medicine
                Signs and Symptoms
                Hypercapnia
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Signs and Symptoms
                Hypercapnia
                Medicine and Health Sciences
                Diagnostic Medicine
                Signs and Symptoms
                Acidosis
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Signs and Symptoms
                Acidosis
                Biology and Life Sciences
                Anatomy
                Nervous System
                Nerves
                Spinal Nerves
                Medicine and Health Sciences
                Anatomy
                Nervous System
                Nerves
                Spinal Nerves
                Biology and Life Sciences
                Anatomy
                Musculoskeletal System
                Muscles
                Abdominal Muscles
                Medicine and Health Sciences
                Anatomy
                Musculoskeletal System
                Muscles
                Abdominal Muscles
                Medicine and Health Sciences
                Anesthesiology
                Anesthesia
                Medicine and Health Sciences
                Pharmaceutics
                Drug Therapy
                Anesthesia
                Biology and Life Sciences
                Anatomy
                Musculoskeletal System
                Muscles
                Pharyngeal Muscles
                Medicine and Health Sciences
                Anatomy
                Musculoskeletal System
                Muscles
                Pharyngeal Muscles
                Research and Analysis Methods
                Mathematical and Statistical Techniques
                Statistical Methods
                Analysis of Variance
                Physical Sciences
                Mathematics
                Statistics (Mathematics)
                Statistical Methods
                Analysis of Variance
                Biology and Life Sciences
                Physiology
                Physiological Processes
                Respiration
                Breathing
                Medicine and Health Sciences
                Physiology
                Physiological Processes
                Respiration
                Breathing
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