GAD67-GFP+ neurons in the Nucleus of Roller: a possible source of inhibitory input to hypoglossal motoneurons. I. Morphology and firing properties.

In this study we examined the electrophysiological and morphological properties of inhibitory neurons located just ventrolateral to the hypoglossal motor (XII) nucleus in the Nucleus of Roller (NR). In vitro experiments were performed on medullary slices derived from postnatal day 5 (P5) to P15 GAD67-GFP knock-in mouse pups. on cell recordings from GFP+ cells in NR in rhythmic slices revealed that these neurons are spontaneously active, although their spiking activity does not exhibit inspiratory phase modulation. Morphologically, GFP+ cells were bi- or multipolar cells with small- to medium-sized cell bodies and small dendritic trees that were often oriented parallel to the border of the XII nucleus. GFP+ cells were classified as either tonic or phasic based on their firing responses to depolarizing step current stimulation in whole cell current clamp. Tonic GFP+ cells fired a regular train of action potentials (APs) throughout the duration of the pulse and often showed rebound spikes after a hyperpolarizing step. In contrast, phasic GFP+ neurons did not fire throughout the depolarizing current step but instead fired fewer than four APs at the onset of the pulse or fired multiple APs, but only after a marked delay. Phasic cells had a significantly smaller input resistance and shorter membrane time constant than tonic GFP+ cells. In addition, phasic GFP+ cells differed from tonic cells in the shape and time course of their spike afterpotentials, the minimum firing frequency at threshold current amplitude, and the slope of their current-frequency relationship. These results suggest that GABAergic neurons in the NR are morphologically and electrophysiologically heterogeneous cells that could provide tonic inhibitory synaptic input to HMs.

[1]  S. Nelson,et al.  Region-Specific Spike-Frequency Acceleration in Layer 5 Pyramidal Neurons Mediated by Kv1 Subunits , 2008, The Journal of Neuroscience.

[2]  D. Bayliss,et al.  Calcium conductances and their role in the firing behavior of neonatal rat hypoglossal motoneurons. , 1993, Journal of neurophysiology.

[3]  N. Tamamaki,et al.  Mechanisms of Neuropeptide Y, Peptide YY, and Pancreatic Polypeptide Inhibition of Identified Green Fluorescent Protein-Expressing GABA Neurons in the Hypothalamic Neuroendocrine Arcuate Nucleus , 2005, The Journal of Neuroscience.

[4]  A. J. Berger,et al.  The Nonuniform Distribution of the GABAA Receptor α1 Subunit Influences Inhibitory Synaptic Transmission to Motoneurons within a Motor Nucleus , 2001, The Journal of Neuroscience.

[5]  P. Nolan,et al.  Suppression of genioglossus muscle tone and activity during reflex hypercapnic stimulation by GABAa mechanisms at the hypoglossal motor nucleus in vivo , 2003, Neuroscience.

[6]  T. Ono,et al.  Swallowing-related Perihypoglossal Neurons Projecting to Hypoglossal Motoneurons in the Cat , 1998, Journal of dental research.

[7]  R. Donato,et al.  Relative contribution by GABA or glycine to Cl(-)-mediated synaptic transmission on rat hypoglossal motoneurons in vitro. , 2000, Journal of neurophysiology.

[8]  Astrid G. Stucke,et al.  Role of inhibitory neurotransmission in the control of canine hypoglossal motoneuron activity in vivo. , 2009, Journal of neurophysiology.

[9]  D A Bayliss,et al.  Repetitive firing properties of developing rat brainstem motoneurones. , 1995, The Journal of physiology.

[10]  S. Prescott,et al.  Integration Time in a Subset of Spinal Lamina I Neurons Is Lengthened by Sodium and Calcium Currents Acting Synergistically to Prolong Subthreshold Depolarization , 2005, The Journal of Neuroscience.

[11]  K. Obata,et al.  Disturbance of neural respiratory control in neonatal mice lacking gaba synthesizing enzyme 67-kda isoform of glutamic acid decarboxylase , 2003, Neuroscience.

[12]  D. Kullmann,et al.  Tonically active GABAA receptors: modulating gain and maintaining the tone , 2004, Trends in Neurosciences.

[13]  Egidio D'Angelo,et al.  Ionic mechanisms of autorhythmic firing in rat cerebellar Golgi cells , 2006, The Journal of physiology.

[14]  R. Horner Respiratory motor activity: influence of neuromodulators and implications for sleep disordered breathing. , 2007, Canadian journal of physiology and pharmacology.

[15]  R. Chronister,et al.  Distribution of glutamic acid decarbocylase and gamma‐aminobutyric acid in thehypoglossal nucleus in the rat , 1988, Journal of neuroscience research.

[16]  G. Barrionuevo,et al.  Morphology of developing rat genioglossal motoneurons studied in vitro: Changes in length, branching pattern, and spatial distribution of dendrites , 1994, The Journal of comparative neurology.

[17]  A. J. Berger,et al.  Gap junctions and inhibitory synapses modulate inspiratory motoneuron synchronization. , 2001, Journal of neurophysiology.

[18]  D. White,et al.  Influence of sleep on genioglossus muscle activation by negative pressure in normal men. , 1993, The American review of respiratory disease.

[19]  D. Richter,et al.  Role of fast inhibitory synaptic mechanisms in respiratory rhythm generation in the maturing mouse. , 1995, The Journal of physiology.

[20]  Yuchio Yanagawa,et al.  Electrophysiological and morphological characteristics of GABAergic respiratory neurons in the mouse pre‐Bötzinger complex , 2006, The European journal of neuroscience.

[21]  J. van Brederode,et al.  GAD67-GFP+ neurons in the Nucleus of Roller. II. Subthreshold and firing resonance properties. , 2011, Journal of neurophysiology.

[22]  J. Feldman,et al.  Dynamic interactions of excitatory and inhibitory inputs in hypoglossal motoneurones: respiratory phasing and modulation by PKA , 2004, The Journal of physiology.

[23]  J. Rekling,et al.  Hypoglossal motoneurons in newborn mice receive respiratory drive from both sides of the medulla , 2009, Neuroscience.

[24]  D. Richter,et al.  Metabotropic glutamate receptors and blockade of glial Krebs cycle depress glycinergic synaptic currents of mouse hypoglossal motoneurons , 2000, The European journal of neuroscience.

[25]  R. Miselis,et al.  Dendritic architecture of hypoglossal motoneurons projecting to extrinsic tongue musculature in the rat , 1994, The Journal of comparative neurology.

[26]  L. Kubin,et al.  Non-reciprocal control of rhythmic activity in respiratory-modulated XII motoneurons. , 1995, Neuroreport.

[27]  J. M. Pattillo,et al.  Role of synaptic inputs in determining input resistance of developing brain stem motoneurons. , 2000, Journal of neurophysiology.

[28]  R. Donato,et al.  Inhibition of spinal or hypoglossal motoneurons of the newborn rat by glycine or GABA , 2002, The European journal of neuroscience.

[29]  George Paxinos,et al.  The Mouse Brain in Stereotaxic Coordinates , 2001 .

[30]  R. Sumino,et al.  Synaptic potentials of hypoglossal motoneurons and a common inhibitory interneuron in the trigemino-hypoglossal reflex. , 1974, Brain research.

[31]  Inhibitory synaptic transmission governs inspiratory motoneuron synchronization. , 2006, Journal of neurophysiology.

[32]  T. Kaneko,et al.  Green fluorescent protein expression and colocalization with calretinin, parvalbumin, and somatostatin in the GAD67‐GFP knock‐in mouse , 2003, The Journal of comparative neurology.

[33]  P. Nolan,et al.  Glycine at hypoglossal motor nucleus: genioglossus activity, CO(2) responses, and the additive effects of GABA. , 2002, Journal of applied physiology.

[34]  N. L. Chamberlin,et al.  Genioglossus premotoneurons and the negative pressure reflex in rats , 2007, The Journal of physiology.

[35]  G. Barrionuevo,et al.  In vitro electrophysiology of developing genioglossal motoneurons in the rat. , 1993, Journal of neurophysiology.

[36]  P. Schwindt,et al.  Multiple potassium conductances and their functions in neurons from cat sensorimotor cortex in vitro. , 1988, Journal of neurophysiology.

[37]  Spike-firing resonance in hypoglossal motoneurons. , 2008, Journal of neurophysiology.

[38]  D A Bayliss,et al.  Multiple potassium conductances and their role in action potential repolarization and repetitive firing behavior of neonatal rat hypoglossal motoneurons. , 1993, Journal of neurophysiology.

[39]  C. Wilson,et al.  Mechanisms Underlying Spontaneous Oscillation and Rhythmic Firing in Rat Subthalamic Neurons , 1999, The Journal of Neuroscience.

[40]  E. Krammer,et al.  Somatotopic organization of the hypoglossal nucleus: a HRP study in the rat , 1979, Brain Research.

[41]  P. Nolan,et al.  GABAA receptor antagonism at the hypoglossal motor nucleus increases genioglossus muscle activity in NREM but not REM sleep , 2003, The Journal of physiology.

[42]  Y. Yanagawa,et al.  GABAergic neurons in inferior colliculus of the GAD67-GFP knock-in mouse: Electrophysiological and morphological properties , 2005, Neuroscience Research.

[43]  J. C. Smith,et al.  Pre-Bötzinger complex: a brainstem region that may generate respiratory rhythm in mammals. , 1991, Science.

[44]  Y. Koninck,et al.  Four cell types with distinctive membrane properties and morphologies in lamina I of the spinal dorsal horn of the adult rat , 2002, The Journal of physiology.

[45]  M. Umemiya,et al.  Presynaptic inhibition by serotonin of glycinergic inhibitory synaptic currents in the rat brain stem. , 1995, Journal of neurophysiology.

[46]  K. Spyer,et al.  Intracellular analysis of respiratory-modulated hypoglossal motoneurons in the cat , 1988, Neuroscience.

[47]  P. Schwindt,et al.  Post‐inhibitory excitation and inhibition in layer V pyramidal neurones from cat sensorimotor cortex. , 1991, The Journal of physiology.

[48]  J. Rekling,et al.  Electrophysiological properties of hypoglossal motoneurons of guinea-pigs studied in vitro , 1989, Neuroscience.

[49]  Y Q Li,et al.  Distribution of GABAergic and glycinergic premotor neurons projecting to the facial and hypoglossal nuclei in the rat , 1997, The Journal of comparative neurology.

[50]  M. Kondo,et al.  Distribution of glycine transporter 2 mRNA-containing neurons in relation to glutamic acid decarboxylase mRNA-containing neurons in rat medulla , 2003, Neuroscience Research.

[51]  Yuchio Yanagawa,et al.  Characterization of GABAergic neurons in rapid‐eye‐movement sleep controlling regions of the brainstem reticular formation in GAD67–green fluorescent protein knock‐in mice , 2008, The European journal of neuroscience.

[52]  H. Kimura,et al.  Immunocytochemical localization of γ‐aminobutyric acid in the hypoglossal nucleus of the macaque monkey, Macaca fuscata: A light and electron microscopic study , 1987 .

[53]  P. H. Hashimoto,et al.  Morphological identification of an interneuron in the hypoglossal nucleus of the rat: A combined Golgi‐electron microscopic study , 1988, The Journal of comparative neurology.

[54]  M. Xi,et al.  Hypoglossal motoneurons are postsynaptically inhibited during carbachol-induced rapid eye movement sleep , 1999, Neuroscience.

[55]  A. J. Berger,et al.  Inspiratory-phase short time scale synchrony in the brainstem slice is generated downstream of the pre-Bötzinger complex , 2008, Neuroscience.