mCLCA3 does not contribute to calcium-activated chloride conductance in murine airways

Lars Mundhenk, Bjarki Johannesson, Pinelopi Anagnostopoulou, Josephine Braun, Melanie K. Bothe, Carsten Schultz, Marcus A. Mall, Achim D. Gruber

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Ca2+-activated Cl- channels (CaCCs) contribute to airway Cl- and fluid secretion, and were implicated in the modulation of disease severity and as a therapeutic target in cystic fibrosis (CF). Previous in vitro studies suggested that members of the CLCA gene family, including the murine mCLCA3, contribute to CaCCs. However, the role of mCLCA3 in ion transport in native airway epithelia has not been studied, to the best of our knowledge. In this study, we used mCLCA3-deficient mice and determined bioelectric properties in freshly excised tracheal tissue, airway morphology, and gene expression studies, to determine the role of mCLCA3 in airway ion transport and airway structure. Bioelectric measurements did not detect any differences in basal short-circuit current, amiloride-sensitive Na+ absorption, cyclic adenosine monophosphate-dependent Cl- secretion, and activation of Ca2+-activated (uridine-5′-triphosphate- mediated) Cl- secretion in mCLCA3-deficient mice compared with wild-type mice. Moreover, no histological changes were observed in the respiratory tract or any other tissues of mCLCA3-deficient mice when compared with wild-type control mice. The intratracheal instillation of IL-13 produced an approximately 30-fold up-regulation of mCLCA3 transcripts without inducing CaCC activity in wild-type airways, and induced goblet-cell hyperplasia and mucin gene expression to similar levels in both genotypes. Further, multiple specific reverse-transcriptase quantitative PCR assays for other CaCC candidates, including mCLCA1, mCLCA2, mCLCA4, mCLCA5, mCLCA6, mCLCA7, mBEST1, mBEST2, mCLC4, mTTYH3, and mTMEM16A, failed to identify the differential expression of genes in the respiratory tract that may compensate for a lack of mCLCA3 function. Together, these findings argue against a role of mCLCA3 in CaCC-mediated Cl - secretion in murine respiratory epithelia.

Original languageEnglish (US)
Pages (from-to)87-93
Number of pages7
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume47
Issue number1
DOIs
StatePublished - Jul 2012
Externally publishedYes

Fingerprint

Calcium Chloride
Chlorides
Calcium
Ion Transport
Gene Expression
Respiratory System
Gene expression
Fluids and Secretions
Genes
Respiratory Mucosa
Uridine Triphosphate
Goblet Cells
Interleukin-13
Ions
Tissue
Amiloride
Mucins
Reverse Transcriptase Polymerase Chain Reaction
Cystic Fibrosis
Uridine

Keywords

  • Airway ion transport
  • CaCC
  • Cl secretion
  • Cystic fibrosis
  • Knockout mouse model

ASJC Scopus subject areas

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

Cite this

mCLCA3 does not contribute to calcium-activated chloride conductance in murine airways. / Mundhenk, Lars; Johannesson, Bjarki; Anagnostopoulou, Pinelopi; Braun, Josephine; Bothe, Melanie K.; Schultz, Carsten; Mall, Marcus A.; Gruber, Achim D.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 47, No. 1, 07.2012, p. 87-93.

Research output: Contribution to journalArticle

Mundhenk, L, Johannesson, B, Anagnostopoulou, P, Braun, J, Bothe, MK, Schultz, C, Mall, MA & Gruber, AD 2012, 'mCLCA3 does not contribute to calcium-activated chloride conductance in murine airways', American Journal of Respiratory Cell and Molecular Biology, vol. 47, no. 1, pp. 87-93. https://doi.org/10.1165/rcmb.2010-0508OC
Mundhenk, Lars ; Johannesson, Bjarki ; Anagnostopoulou, Pinelopi ; Braun, Josephine ; Bothe, Melanie K. ; Schultz, Carsten ; Mall, Marcus A. ; Gruber, Achim D. / mCLCA3 does not contribute to calcium-activated chloride conductance in murine airways. In: American Journal of Respiratory Cell and Molecular Biology. 2012 ; Vol. 47, No. 1. pp. 87-93.
@article{b0fe311fe9e04d089aea3d97ca3d88d9,
title = "mCLCA3 does not contribute to calcium-activated chloride conductance in murine airways",
abstract = "Ca2+-activated Cl- channels (CaCCs) contribute to airway Cl- and fluid secretion, and were implicated in the modulation of disease severity and as a therapeutic target in cystic fibrosis (CF). Previous in vitro studies suggested that members of the CLCA gene family, including the murine mCLCA3, contribute to CaCCs. However, the role of mCLCA3 in ion transport in native airway epithelia has not been studied, to the best of our knowledge. In this study, we used mCLCA3-deficient mice and determined bioelectric properties in freshly excised tracheal tissue, airway morphology, and gene expression studies, to determine the role of mCLCA3 in airway ion transport and airway structure. Bioelectric measurements did not detect any differences in basal short-circuit current, amiloride-sensitive Na+ absorption, cyclic adenosine monophosphate-dependent Cl- secretion, and activation of Ca2+-activated (uridine-5′-triphosphate- mediated) Cl- secretion in mCLCA3-deficient mice compared with wild-type mice. Moreover, no histological changes were observed in the respiratory tract or any other tissues of mCLCA3-deficient mice when compared with wild-type control mice. The intratracheal instillation of IL-13 produced an approximately 30-fold up-regulation of mCLCA3 transcripts without inducing CaCC activity in wild-type airways, and induced goblet-cell hyperplasia and mucin gene expression to similar levels in both genotypes. Further, multiple specific reverse-transcriptase quantitative PCR assays for other CaCC candidates, including mCLCA1, mCLCA2, mCLCA4, mCLCA5, mCLCA6, mCLCA7, mBEST1, mBEST2, mCLC4, mTTYH3, and mTMEM16A, failed to identify the differential expression of genes in the respiratory tract that may compensate for a lack of mCLCA3 function. Together, these findings argue against a role of mCLCA3 in CaCC-mediated Cl - secretion in murine respiratory epithelia.",
keywords = "Airway ion transport, CaCC, Cl secretion, Cystic fibrosis, Knockout mouse model",
author = "Lars Mundhenk and Bjarki Johannesson and Pinelopi Anagnostopoulou and Josephine Braun and Bothe, {Melanie K.} and Carsten Schultz and Mall, {Marcus A.} and Gruber, {Achim D.}",
year = "2012",
month = "7",
doi = "10.1165/rcmb.2010-0508OC",
language = "English (US)",
volume = "47",
pages = "87--93",
journal = "American Journal of Respiratory Cell and Molecular Biology",
issn = "1044-1549",
publisher = "American Thoracic Society",
number = "1",

}

TY - JOUR

T1 - mCLCA3 does not contribute to calcium-activated chloride conductance in murine airways

AU - Mundhenk, Lars

AU - Johannesson, Bjarki

AU - Anagnostopoulou, Pinelopi

AU - Braun, Josephine

AU - Bothe, Melanie K.

AU - Schultz, Carsten

AU - Mall, Marcus A.

AU - Gruber, Achim D.

PY - 2012/7

Y1 - 2012/7

N2 - Ca2+-activated Cl- channels (CaCCs) contribute to airway Cl- and fluid secretion, and were implicated in the modulation of disease severity and as a therapeutic target in cystic fibrosis (CF). Previous in vitro studies suggested that members of the CLCA gene family, including the murine mCLCA3, contribute to CaCCs. However, the role of mCLCA3 in ion transport in native airway epithelia has not been studied, to the best of our knowledge. In this study, we used mCLCA3-deficient mice and determined bioelectric properties in freshly excised tracheal tissue, airway morphology, and gene expression studies, to determine the role of mCLCA3 in airway ion transport and airway structure. Bioelectric measurements did not detect any differences in basal short-circuit current, amiloride-sensitive Na+ absorption, cyclic adenosine monophosphate-dependent Cl- secretion, and activation of Ca2+-activated (uridine-5′-triphosphate- mediated) Cl- secretion in mCLCA3-deficient mice compared with wild-type mice. Moreover, no histological changes were observed in the respiratory tract or any other tissues of mCLCA3-deficient mice when compared with wild-type control mice. The intratracheal instillation of IL-13 produced an approximately 30-fold up-regulation of mCLCA3 transcripts without inducing CaCC activity in wild-type airways, and induced goblet-cell hyperplasia and mucin gene expression to similar levels in both genotypes. Further, multiple specific reverse-transcriptase quantitative PCR assays for other CaCC candidates, including mCLCA1, mCLCA2, mCLCA4, mCLCA5, mCLCA6, mCLCA7, mBEST1, mBEST2, mCLC4, mTTYH3, and mTMEM16A, failed to identify the differential expression of genes in the respiratory tract that may compensate for a lack of mCLCA3 function. Together, these findings argue against a role of mCLCA3 in CaCC-mediated Cl - secretion in murine respiratory epithelia.

AB - Ca2+-activated Cl- channels (CaCCs) contribute to airway Cl- and fluid secretion, and were implicated in the modulation of disease severity and as a therapeutic target in cystic fibrosis (CF). Previous in vitro studies suggested that members of the CLCA gene family, including the murine mCLCA3, contribute to CaCCs. However, the role of mCLCA3 in ion transport in native airway epithelia has not been studied, to the best of our knowledge. In this study, we used mCLCA3-deficient mice and determined bioelectric properties in freshly excised tracheal tissue, airway morphology, and gene expression studies, to determine the role of mCLCA3 in airway ion transport and airway structure. Bioelectric measurements did not detect any differences in basal short-circuit current, amiloride-sensitive Na+ absorption, cyclic adenosine monophosphate-dependent Cl- secretion, and activation of Ca2+-activated (uridine-5′-triphosphate- mediated) Cl- secretion in mCLCA3-deficient mice compared with wild-type mice. Moreover, no histological changes were observed in the respiratory tract or any other tissues of mCLCA3-deficient mice when compared with wild-type control mice. The intratracheal instillation of IL-13 produced an approximately 30-fold up-regulation of mCLCA3 transcripts without inducing CaCC activity in wild-type airways, and induced goblet-cell hyperplasia and mucin gene expression to similar levels in both genotypes. Further, multiple specific reverse-transcriptase quantitative PCR assays for other CaCC candidates, including mCLCA1, mCLCA2, mCLCA4, mCLCA5, mCLCA6, mCLCA7, mBEST1, mBEST2, mCLC4, mTTYH3, and mTMEM16A, failed to identify the differential expression of genes in the respiratory tract that may compensate for a lack of mCLCA3 function. Together, these findings argue against a role of mCLCA3 in CaCC-mediated Cl - secretion in murine respiratory epithelia.

KW - Airway ion transport

KW - CaCC

KW - Cl secretion

KW - Cystic fibrosis

KW - Knockout mouse model

UR - http://www.scopus.com/inward/record.url?scp=84863808527&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84863808527&partnerID=8YFLogxK

U2 - 10.1165/rcmb.2010-0508OC

DO - 10.1165/rcmb.2010-0508OC

M3 - Article

VL - 47

SP - 87

EP - 93

JO - American Journal of Respiratory Cell and Molecular Biology

JF - American Journal of Respiratory Cell and Molecular Biology

SN - 1044-1549

IS - 1

ER -