P2X2 receptors differentiate placodal vs. neural crest C-fiber phenotypes innervating guinea pig lungs and esophagus.

The lungs and esophagus are innervated by sensory neurons with somata in the nodose, jugular, and dorsal root ganglion. These sensory ganglia are derived from embryonic placode (nodose) and neural crest tissues (jugular and dorsal root ganglia; DRG). We addressed the hypothesis that the neuron's embryonic origin (e.g., placode vs. neural crest) plays a greater role in determining particular aspects of its phenotype than the environment in which it innervates (e.g., lungs vs. esophagus). This hypothesis was tested using a combination of extracellular and patch-clamp electrophysiology and single-cell RT-PCR from guinea pig neurons. Nodose, but not jugular C-fibers innervating the lungs and esophagus, responded to alpha,beta-methylene ATP with action potential discharge that was sensitive to the P2X3 (P2X2/3) selective receptor antagonist A-317491. The somata of lung- and esophagus-specific sensory fibers were identified using retrograde tracing with a fluorescent dye. Esophageal- and lung-traced neurons from placodal tissue (nodose neurons) responded similarly to alpha,beta-methylene ATP (30 microM) with a large sustained inward current, whereas in neurons derived from neural crest tissue (jugular and DRG neurons), the same dose of alpha,beta-methylene ATP resulted in only a transient rapidly inactivating current or no detectable current. It has been shown previously that only activation of P2X2/3 heteromeric receptors produce sustained currents, whereas homomeric P2X3 receptor activation produces a rapidly inactivating current. Consistent with this, single-cell RT-PCR analysis revealed that the nodose ganglion neurons innervating the lungs and esophagus expressed mRNA for P2X2 and P2X3 subunits, whereas the vast majority of jugular and dorsal root ganglia innervating these tissues expressed only P2X3 mRNA with little to no P2X2 mRNA expression. We conclude that the responsiveness of C-fibers innervating the lungs and esophagus to ATP and other purinergic agonists is determined more by their embryonic origin than by the environment of the tissue they ultimately innervate.

[1]  K. Kwong,et al.  Expression and function of the ion channel TRPA1 in vagal afferent nerves innervating mouse lungs , 2008, The Journal of physiology.

[2]  K. Kwong,et al.  Voltage‐gated sodium channels in nociceptive versus non‐nociceptive nodose vagal sensory neurons innervating guinea pig lungs , 2008, The Journal of physiology.

[3]  M. Kollarik,et al.  Evidence for both adenosine A1 and A2A receptors activating single vagal sensory C‐fibres in guinea pig lungs , 2006, The Journal of physiology.

[4]  D. Weinreich,et al.  Reflex regulation of airway sympathetic nerves in guinea‐pigs , 2006, The Journal of physiology.

[5]  W. Snider,et al.  “Runx”ing towards Sensory Differentiation , 2006, Neuron.

[6]  D. Cockayne,et al.  P2X2 knockout mice and P2X2/P2X3 double knockout mice reveal a role for the P2X2 receptor subunit in mediating multiple sensory effects of ATP , 2005, The Journal of physiology.

[7]  M. Kollarik,et al.  Effect of 5-hydroxytryptamine on vagal C-fiber subtypes in guinea pig lungs. , 2005, Pulmonary pharmacology & therapeutics.

[8]  D. Weinreich,et al.  Advances in Vagal Afferent Neurobiology , 2005 .

[9]  M. Kollarik,et al.  Vagal afferent nerves with nociceptive properties in guinea‐pig oesophagus , 2005, The Journal of physiology.

[10]  S. Meeker,et al.  Identification of the tracheal and laryngeal afferent neurones mediating cough in anaesthetized guinea‐pigs , 2004, The Journal of physiology.

[11]  D. Weinreich,et al.  Subtypes of vagal afferent C‐fibres in guinea‐pig lungs , 2004, The Journal of physiology.

[12]  Darrell R. Abernethy,et al.  International Union of Pharmacology: Approaches to the Nomenclature of Voltage-Gated Ion Channels , 2003, Pharmacological Reviews.

[13]  R. North,et al.  Subunit Arrangement in P2X Receptors , 2003, The Journal of Neuroscience.

[14]  M. Kollarik,et al.  Capsaicin‐sensitive and ‐insensitive vagal bronchopulmonary C‐fibres in the mouse , 2003, The Journal of physiology.

[15]  T. Brennan,et al.  A-317491, a novel potent and selective non-nucleotide antagonist of P2X3 and P2X2/3 receptors, reduces chronic inflammatory and neuropathic pain in the rat , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[16]  G. Burnstock,et al.  Localization of P2X3 receptors and coexpression with P2X2 receptors during rat embryonic neurogenesis , 2002, The Journal of comparative neurology.

[17]  G. Burnstock,et al.  P2x Receptors in Peripheral Neurons , 2000 .

[18]  B S Khakh,et al.  International union of pharmacology. XXIV. Current status of the nomenclature and properties of P2X receptors and their subunits. , 2001, Pharmacological reviews.

[19]  L. Lee,et al.  Afferent properties and reflex functions of bronchopulmonary C-fibers. , 2001, Respiration physiology.

[20]  G Burnstock,et al.  Diinosine pentaphosphate: an antagonist which discriminates between recombinant P2X3 and P2X2/3 receptors and between two P2X receptors in rat sensory neurones , 2000, British journal of pharmacology.

[21]  R. North,et al.  Contribution of individual subunits to the multimeric P2X(2) receptor: estimates based on methanethiosulfonate block at T336C. , 1999, Molecular pharmacology.

[22]  C. Stucky,et al.  Isolectin B4-Positive and -Negative Nociceptors Are Functionally Distinct , 1999, The Journal of Neuroscience.

[23]  B. Undem,et al.  Identification and substance P content of vagal afferent neurons innervating the epithelium of the guinea pig trachea. , 1999, American journal of respiratory and critical care medicine.

[24]  A. Nicke,et al.  P2X1 and P2X3 receptors form stable trimers: a novel structural motif of ligand‐gated ion channels , 1998, The EMBO journal.

[25]  R. North,et al.  Immunohistochemical study of the P2X2 and P2X3 receptor subunits in rat and monkey sensory neurons and their central terminals , 1997, Neuropharmacology.

[26]  B. Undem,et al.  Immunomodulation of afferent neurons in guinea‐pig isolated airway. , 1996, The Journal of physiology.

[27]  M. Radeke,et al.  Presence or absence of TrKA protein distinguishes subsets of small sensory neurons with unique cytochemical characteristics and dorsal horn projections , 1995, The Journal of comparative neurology.

[28]  R. North,et al.  Coexpression of P2X2 and P2X3 receptor subunits can account for ATP-gated currents in sensory neurons , 1995, Nature.

[29]  J. Polak,et al.  Retrograde tracing shows that CGRP-immunoreactive nerves of rat trachea and lung originate from vagal and dorsal root ganglia. , 1987, Journal of the autonomic nervous system.

[30]  S. Hunt,et al.  Peptide- and non-peptide-containing unmyelinated primary afferents: the parallel processing of nociceptive information. , 1985, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[31]  G. Sant'ambrogio,et al.  Circulatory accessibility of nervous receptors localized in the tracheobronchial tree. , 1982, Respiration physiology.

[32]  Y. Jammes,et al.  Afferent and efferent components of the bronchial vagal branches in cats. , 1982, Journal of the autonomic nervous system.

[33]  H. Coleridge,et al.  Impulse activity in afferent vagal C-fibres with endings in the intrapulmonary airways of dogs. , 1977, Respiration physiology.

[34]  A. Paintal,et al.  Mechanism of stimulation of type J pulmonary receptors , 1969, The Journal of physiology.

[35]  E. Agostoni,et al.  Functional and histological studies of the vagus nerve and its branches to the heart, lungs and abdominal viscera in the cat , 1957, The Journal of physiology.

[36]  B. A. Paintal Impulses in vagal afferent fibres from specific pulmonary deflation receptors: the response of these receptors to phenyl diguanide, potato starch, 5-hydroxytryptamine and nicotine, and their rôle in respiratory and cardiovascular reflexes. , 1955, Quarterly journal of experimental physiology and cognate medical sciences.

[37]  A. Paintal The response of gastric stretch receptors and certain other abdominal and thoracic vagal receptors to some drugs , 1954, The Journal of physiology.

[38]  C. Baker,et al.  The Embryology of Vagal Sensory Neurons , 2006 .

[39]  H. Coleridge,et al.  Afferent vagal C fibre innervation of the lungs and airways and its functional significance. , 1984, Reviews of physiology, biochemistry and pharmacology.

[40]  A. Paintal,et al.  Vagal sensory receptors and their reflex effects. , 1973, Physiological reviews.