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...Continue Reading
Citations
Sep 24, 1998·Trends in Pharmacological Sciences· Haddad el-B, J Rousell
Mar 27, 2003·Pulmonary Pharmacology & Therapeutics·P J KinghamW Graham McLean
Sep 23, 1997·British Journal of Pharmacology·A F RoffelH Kilbinger
May 14, 1998·British Journal of Pharmacology·Y MuraiY Ito
Jul 6, 2000·British Journal of Pharmacology·D A KnightP J Thompson
May 22, 1998·Journal of Cardiovascular Pharmacology·L B FernandesR G Goldie
Nov 3, 1998·Thorax·R W CostelloA D Fryer
May 26, 2009·American Journal of Physiology. Lung Cellular and Molecular Physiology·Liliana Moreno-VinascoDavid B Jacoby
Apr 18, 2001·American Journal of Respiratory Cell and Molecular Biology·D B JacobyA D Fryer
Jul 3, 1998·The Journal of Clinical Investigation·D B JacobyA D Fryer
Dec 31, 1997·The Journal of Clinical Investigation·C M EvansR W Costello
Dec 24, 2005·The Journal of Clinical Investigation·Allison D FryerDavid B Jacoby
Jan 5, 2011·British Journal of Pharmacology·Bart C Moulton, Allison D Fryer
Jun 29, 2004·Pulmonary Pharmacology & Therapeutics·K Racké, S Matthiesen
Sep 13, 2000·Journal of Smooth Muscle Research = Nihon Heikatsukin Gakkai Kikanshi·T NakajimaY Ito
Aug 6, 2004·Journal of Aerosol Medicine : the Official Journal of the International Society for Aerosols in Medicine·David B Jacoby
Jul 28, 1999·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·D B Jacoby, A D Fryer
Jan 6, 2004·American Journal of Physiology. Lung Cellular and Molecular Physiology·Allison D FryerDavid A Jett
Sep 2, 2003·American Journal of Physiology. Lung Cellular and Molecular Physiology·Deborah A SawatzkyRichard W Costello
May 11, 2002·American Journal of Physiology. Lung Cellular and Molecular Physiology·Deborah A SawatzkyRichard W Costello