Cellular Distribution and Subcellular Localization of mCLCA1/2 in Murine Gastrointestinal Epithelia

Roussa, Eleni; Wittschen, Petra; Wolff, Natascha A.; Torchalski, Blazej; Gruber, Achim D.; Thévenod, Frank

doi:10.1369/jhc.2010.955211

Cited by 14 publications

(10 citation statements)

References 35 publications

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“…Our list of DEGs includes those highly relevant to intracellular signaling and ion transport. In keeping with previous reports that chloride deficiencies affect enamel growth (21)(22)(23), the levels of the Clca2 transcript, encoding an anion channel protein putatively involved in chloride transport, protein secretion, and cell adhesion (24,25), were more abundant in the midto late stage of maturation in both experimental groups (10.2-and 8.5-fold up-regulation, respectively). CLCA2 proteins are commonly localized in the plasma membranes of secretory epithelial cells (26,27).…”

Section: Discussionsupporting

confidence: 90%

Gene‐expression analysis of early‐ and late‐maturation‐stage rat enamel organ

Lacruz

Smith

Chen

et al. 2011

European J Oral Sciences

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Enamel maturation is a dynamic process that involves high rates of mineral acquisition, associated fluctuations in extracellular pH and resorption of extracellular enamel proteins. During maturation, ameloblasts change from a tall, thin and highly polarized organization characteristic of the secretory stage, to a low columnar and widened morphology in the maturation stage. To identify potential differences in gene expression throughout maturation, we obtained enamel organ epithelial cells derived from the early and late maturation stages from rat incisor and analyzed global gene expression profiles at each stage. Sixty three candidate genes were identified with potential roles in the maturation process. qPCR was used to confirm results from this genome-wide analysis in a subset of genes. Enriched transcripts in late maturation (n= 38) included those associated with lysosomal activity, solute carrier transport and calcium signaling. Cellular responses to oxidative stress, proton transport, cell death and immune system-related transcripts were also up-regulated. Transcripts down-regulated in the late maturation stage (n= 25) included those with functions related to cell adhesion, cell signaling, and T-cell activation. These results indicate that ameloblasts undergo widespread molecular changes during the maturation stage of amelogenesis and so provide the bases for future functional investigations into the mechanistic basis of enamel mineralization.

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Section: Discussionsupporting

confidence: 90%

Gene‐expression analysis of early‐ and late‐maturation‐stage rat enamel organ

Lacruz

Smith

Chen

et al. 2011

European J Oral Sciences

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“…For single stainings, slides of murine lungs were incubated with immunopurified rabbit antibodies for mCLCA5 (α-mCLCA5-C1-ap, 1:300) (Braun et al 2010 ), mCLCA3 (α-m3-C-1p, 1:600) (Bothe et al 2011 ) or cytokeratin 5 (1:1,000; ab24647, Abcam) or with the immunopurified goat antibody for club cell 10 protein (CC10, 1:1,500; sc-9772, Santa Cruz Biotechnology). To exclude cross-reactivity of the mCLCA5 antibody with other murine CLCA members, immunohistochemical stainings of the intestine, expressing mCLCA3, mCLCA4, mCLCA6 and mCLCA7 (Patel et al 2009 ), and the pancreas, expressing mCLCA1/2 (Roussa et al 2010 ), were performed and yielded negative results. Furthermore, specificity of the mCLCA5 antibody was previously verified by immunoblot analysis (Braun et al 2010 ).…”

Section: Methodsmentioning

confidence: 99%

Murine CLCA5 is uniquely expressed in distinct niches of airway epithelial cells

Dietert

Mundhenk

Erickson

et al. 2014

Histochem Cell Biol

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The murine mCLCA5 protein is a member of the chloride channel regulators, calcium-activated (CLCA) family and is suspected to play a role in airway mucus cell differentiation. Although mCLCA5 mRNA was previously found in total lung extracts, the expressing cells and functions in the naive murine respiratory tract are unknown. Therefore, mCLCA5 protein expression was identified by immunohistochemistry and confocal laser scanning microscopy using entire lung sections of naive mice. Moreover, we determined mRNA levels of functionally related genes (mClca3, mClca5, Muc5ac and Muc5b) and quantified mCLCA5-, mCLCA3- and CC10-positive cells and periodic acid-Schiff-positive mucus cells in naive, PBS-treated or Staphylococcus aureus-infected mice. We also investigated mCLCA5 protein expression in Streptococcus pneumoniae and influenza virus lung infection models. Finally, we determined species-specific differences in the expression patterns of the murine mCLCA5 and its human and porcine orthologs, hCLCA2 and pCLCA2. The mCLCA5 protein is uniquely expressed in highly select bronchial epithelial cells and submucosal glands in naive mice, consistent with anatomical locations of progenitor cell niches. Under conditions of challenge (PBS, S. aureus, S. pneumoniae, influenza virus), mRNA and protein expression strongly declined with protein recovery only in models retaining intact epithelial cells. In contrast to mice, human and porcine bronchial epithelial cells do not express their respective mCLCA5 orthologs and submucosal glands had fewer expressing cells, indicative of fundamental differences in mice versus humans and pigs.

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“…Of note, CLCA3a1 and CLCA3a2 share 96% nucleic acid sequence identity on the cDNA level and 92% on the amino acid level [ 19 , 23 ], raising the question of whether some of the expression data may have resulted from unintended cross-reactivity of the probes used. Not surprisingly, available antibodies failed to discriminate between these two homologs [ 24 ]. Only a real–time RT-PCR approach using taqman-probes reliably discriminated between CLCA3a1 which is predominantly expressed in spleen and bone marrow and CLCA3a2 which was found in the lactating and involuting mammary gland [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

Interspecies diversity of chloride channel regulators, calcium-activated 3 genes

et al. 2018

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Members of the chloride channel regulators, calcium-activated (CLCA) family, have been implicated in diverse biomedical conditions, including chronic inflammatory airway diseases such as asthma, chronic obstructive pulmonary disease, and cystic fibrosis, the activation of macrophages, and the growth and metastatic spread of tumor cells. Several observations, however, could not be repeated across species boundaries and increasing evidence suggests that select CLCA genes are particularly prone to dynamic species-specific evolvements. Here, we systematically characterized structural and expressional differences of the CLCA3 gene across mammalian species, revealing a spectrum of gene duplications, e.g., in mice and cows, and of gene silencing via diverse chromosomal modifications in pigs and many primates, including humans. In contrast, expression of a canonical CLCA3 protein from a single functional gene seems to be evolutionarily retained in carnivores, rabbits, guinea pigs, and horses. As an accepted asthma model, we chose the cat to establish the tissue and cellular expression pattern of the CLCA3 protein which was primarily found in mucin-producing cells of the respiratory tract and in stratified epithelia of the esophagus. Our results suggest that, among developmental differences in other CLCA genes, the CLCA3 gene possesses a particularly high dynamic evolutionary diversity with pivotal consequences for humans and other primates that seem to lack a CLCA3 protein. Our data also help to explain previous contradictory results on CLCA3 obtained from different species and warrant caution in extrapolating data from animal models in conditions where CLCA3 may be involved.

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Cellular Distribution and Subcellular Localization of mCLCA1/2 in Murine Gastrointestinal Epithelia

Cited by 14 publications

References 35 publications

Gene‐expression analysis of early‐ and late‐maturation‐stage rat enamel organ

Gene‐expression analysis of early‐ and late‐maturation‐stage rat enamel organ

Murine CLCA5 is uniquely expressed in distinct niches of airway epithelial cells

Interspecies diversity of chloride channel regulators, calcium-activated 3 genes

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