American Journal of Respiratory Cell and Molecular Biology. Vol. 28, pp. 473-477, 2003
© 2003 American Thoracic Society DOI: 10.1165/rcmb.2002-0190OC
Expression of the High-Affinity Choline Transporter, CHT1, in the Rat Trachea
Uwe Pfeil,
Katrin Susanne Lips,
Lars Eberling,
Veronika Grau,
Rainer Viktor Haberberger and
Wolfgang Kummer
Institutes for Anatomy and Cell Biology, Justus-Liebig-University, Giessen; and Philipps-University, Marburg, Germany
Address correspondence to: Dr. Uwe Pfeil, Institute for Anatomy and Cell Biology, Aulweg 123, 35385 Giessen, Germany. E-mail: uwe.pfeil{at}anatomie.med.uni-giessen.de
The rate limiting step in neuronal acetylcholine (ACh) synthesis is the uptake of choline by the high-affinity choline transporter (CHT1). Here, we investigated the distribution of CHT1 in the rat trachea. CHT1-mRNA was detected by reverse transcriptasepolymerase chain reaction in trachea without epithelium, abraded tracheal mucosa, and in epithelial cells obtained by laser-assisted cell-picking. Accordingly, CHT1-mRNA could also be detected in tracheal epithelial cells by in situ hybridization. Recently obtained polyclonal rabbit and guinea-pig antisera against a synthetic peptide corresponding to amino acid residues 2940 of the rat CHT1 sequence localized CHT1 protein in combination with antisera against the vesicular acetylcholine transporter in cholinergic fibers innervating tracheal glands and the tracheal muscle. In case of the tracheal epithelium, CHT1 was restricted to the apical membrane of the ciliated cells, as demonstrated by confocal laser scanning and electron microscopy using an affinity-purified CHT1 antiserum. The close apposition of CHT1 to reported sites of localization of choline acetyltransferase in these cells is strongly in favor of ACh synthesis being fuelled by choline uptake via CHT1 after release and breakdown of ACh at the luminal surface. Accordingly, cholinergic regulation of tracheal epithelial function is governed by local release and recycling of ACh by ciliated cells.
Abbreviations: acetylcholine, ACh high-affinity choline transporter, CHT choline acetyltransferase, ChAT digoxygenin, DIG polymerase chain reaction, PCR reverse transcriptase, RT Tris-buffered saline with 0.05% Tween-20, TTBS vesicular acetylcholine transporter, VAChT
This article has been cited by other articles:

|
 |

|
 |
 
A. Hecker, Z. Mikulski, K. S. Lips, U. Pfeil, A. Zakrzewicz, S. Wilker, P. Hartmann, W. Padberg, I. Wessler, W. Kummer, et al.
Pivotal Advance: Up-regulation of acetylcholine synthesis and paracrine cholinergic signaling in intravascular transplant leukocytes during rejection of rat renal allografts
J. Leukoc. Biol.,
July 1, 2009;
86(1):
13 - 22.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
V. Michel, Z. Yuan, S. Ramsubir, and M. Bakovic
Choline Transport for Phospholipid Synthesis.
Experimental Biology and Medicine,
May 1, 2006;
231(5):
490 - 504.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. D. Fullerton, L. Wagner, Z. Yuan, and M. Bakovic
Impaired trafficking of choline transporter-like protein-1 at plasma membrane and inhibition of choline transport in THP-1 monocyte-derived macrophages
Am J Physiol Cell Physiol,
April 1, 2006;
290(4):
C1230 - C1238.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
K. S. Lips, C. Volk, B. M. Schmitt, U. Pfeil, P. Arndt, D. Miska, L. Ermert, W. Kummer, and H. Koepsell
Polyspecific Cation Transporters Mediate Luminal Release of Acetylcholine from Bronchial Epithelium
Am. J. Respir. Cell Mol. Biol.,
July 1, 2005;
33(1):
79 - 88.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. Xie and Q. Guo
Par-4 Inhibits Choline Uptake by Interacting with CHT1 and Reducing Its Incorporation on the Plasma Membrane
J. Biol. Chem.,
July 2, 2004;
279(27):
28266 - 28275.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
B. J. Proskocil, H. S. Sekhon, Y. Jia, V. Savchenko, R. D. Blakely, J. Lindstrom, and E. R. Spindel
Acetylcholine Is an Autocrine or Paracrine Hormone Synthesized and Secreted by Airway Bronchial Epithelial Cells
Endocrinology,
May 1, 2004;
145(5):
2498 - 2506.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
K. S. Lips, U. Pfeil, K. Reiners, C. Rimasch, K. Kuchelmeister, R. C. Braun-Dullaeus, R. V. Haberberger, R. Schmidt, and W. Kummer
Expression of the High-affinity Choline Transporter CHT1 in Rat and Human Arteries
J. Histochem. Cytochem.,
December 1, 2003;
51(12):
1645 - 1654.
[Abstract]
[Full Text]
[PDF]
|
 |
|
Copyright © 2003 American Thoracic Society.
|
|
|