Cultures of Airway Parasympathetic Nerves Express Functional M2 Muscarinic Receptors

Allison D. Fryer, Constance L. Elbon, Amanda L. Kim, Hui Qing Xiao, Allan I. Levey, David B. Jacoby

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

To study the control of acetylcholine release from airway parasympathetic neurons, primary cultures of these cells were established. Guinea pig tracheas were disaggregated with collagenase and plated onto matrigel-coated plates in medium that contained cytosine arabinoside to inhibit growth of dividing cells. Over 7 to 10 days neurites grow from the cell bodies, reaching a length of 2 mm. The vast majority of the cells in these cultures were neurons, as identified by morphology and staining with Neurotag® and with antibody to neuron-specific antigen protein gene product 9.5. Cultured neurons contained acetylcholine, which was released by electrical field stimulation. Thus these were parasympathetic neurons. Staining with antibodies to M1, M2, and M4 muscarinic receptors revealed the presence of only M2 receptors. Likewise, reverse transcription-polymerase chain reaction using primers for M1, M2, and M4 muscarinic receptors revealed mRNA only for M2 receptors. Blocking these M2 receptors using atropine potentiated the stimulated release of acetylcholine, demonstrating that the M2 receptors inhibit acetylcholine release, as they have been shown to do in vivo. Thus airway parasympathetic neurons can be grown in culture, they retain the ability to synthesize and release acetylcholine, and they express functional inhibitory M2 muscarinic receptors.

Original languageEnglish (US)
Pages (from-to)716-725
Number of pages10
JournalAmerican journal of respiratory cell and molecular biology
Volume15
Issue number6
DOIs
StatePublished - Jan 1 1996

    Fingerprint

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this