Immunohistochemical Localization of Cystic Fibrosis Transmembrane Conductance Regulator in Human Fetal Airway and Digestive Mucosa

Gaillard, Dominique; Ruocco, S M; Lallemand, A; Dalemans, Wilfried; Hinnrasky, Jocelyne; Puchelle, Édith

doi:10.1203/00006450-199408000-00002

Cited by 40 publications

(30 citation statements)

References 8 publications

(8 reference statements)

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“…CFTR in + /AF tissues had a similar localization (not shown). Expression in these tissues agrees with previous studies using in situ hybridization and immunocytochemistry of normal rodent and human tissue (30)(31)(32). In AF/AF and null mice, the Brunner's glands were often dilated (Fig.…”

Section: Methodssupporting

confidence: 90%

A mouse model for the delta F508 allele of cystic fibrosis.

et al. 1995

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The most common cause of cystic fibrosis is a mutation that deletes phenylalanine 508 in cystic fibrosis transmembrane conductance regulator (CFTR). The AF508 protein is misprocessed and degraded rather than traveling to the apical membrane. We used a novel strategy to introduce the AF508 mutation into the mouse CFTR gene. Affected epithelia from homozygous AF508 mice lacked CFTR in the apical membrane and were Cl -impermeable. These abnormalities are the same as those observed in patients with AF508 and suggest that these mice have the same cellular defect. 40% of homozygous AF508 animals survived into adulthood and displayed several abnormalities found in human disease and in CFTR null mice. These animals should provide an excellent model to investigate pathogenesis and to examine therapies directed at correcting the AF508 defect. (J. Clin.

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Section: Methodssupporting

confidence: 90%

A mouse model for the delta F508 allele of cystic fibrosis.

et al. 1995

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“…3), we observe that CFTR is in the AR of TCE cells, both ciliated and nonciliated (the latter comprising goblet cells). This confirms previous findings of immunocytochemical studies on fetal airways (Gaillard et al, 1994) and nasal polyps (Puchelle et al, 1992). However, it contradicts other immunocytochemical observations of the human bronchus, which were unable to detect CFTR in ciliated cells (perhaps using a less sensitive method), and restricting it to the submucosal glands (Engelhardt et al, 1992;.…”

Section: Localization Of Cftr In Nasal Brushing Cellssupporting

confidence: 86%

Cystic Fibrosis F508del Patients Have Apically Localized CFTR in a Reduced Number of Airway Cells

Penque

Mendes

Beck

et al. 2000

Laboratory Investigation

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SUMMARY:Present state of knowledge, mostly based on heterologous expression studies, indicates that the cystic fibrosis transmembrane conductance regulator (CFTR) protein bearing the F508del mutation is misprocessed and mislocalized in the cytoplasm, unable to reach the cell surface. Recently, however, it was described that protein levels and localization are similar between F508del and wild-type CFTR in airway and intestinal tissues, but not in the sweat glands. In this study, we used immunocytochemistry with three different anti-CFTR antibodies to investigate endogenous CFTR expression and localization in nasal epithelial cells from F508del homozygous patients, F508del carriers, and non-CF individuals. On average, 300 cells were observed per individual. No significant differences were observed for cell type distributions among CF, carrier, and non-CF samples; epithelial cells made up approximately 80% to 95% of all cells present. CFTR was detected mostly in the apical region (AR) of the tall columnar epithelial (TCE) cells, ciliated or nonciliated. By confocal microscopy analysis, we show that the CFTR apical region-staining does not overlap with either anti-calnexin (endoplasmic reticulum), anti-p58 (Golgi), or anti-tubulin (cilia) stainings. The median from results with three antibodies indicate that the apical localization of CFTR happens in 22% of TCE cells from F508del homozygous patients with CF (n ϭ 12), in 42% of cells from F508del carriers (n ϭ 20), and in 56% of cells from healthy individuals (n ϭ 12). Statistical analysis indicates that differences are significant among all groups studied and for the three antibodies (p Ͻ 0.05). These results confirm the presence of CFTR in the apical region of airway cells from F508del homozygous patients; however, they also reveal that the number of cells in which this occurs is significantly lower than in F508del carriers and much lower than in healthy individuals. These findings may have an impact on the design of novel pharmacological strategies aimed at circumventing the CF defect caused by the F508del mutation. (Lab Invest 2000, 80:857-868).

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“…In airway tissue, the expression of CFTR mRNA transcripts in undifferentiated fetal epithelium appears to be very low (34). CFTR protein expression is localized in the apical domain of the ciliated cells by the 7th week of gestation (5). Interestingly, it has been reported that in rat regenerating liver the quantitative levels of CFTR gene activity is the result of transcriptional and posttranscriptional phenomena (35).…”

Section: Discussionmentioning

confidence: 99%

“…Oligodeoxynucleotides were synthesized using the phosphoamidite method. The following primers were made for the detection of mRNA transcripts: aldolase sense primer oald 141N 5'-GGCAAGGGCATCCTGGCT-GCAGA, aldolase antisense primer oaldS83T 5 The score of inflammation was qualitatively evaluated on HPSstained tissue sections by analyzing the presence of inflammatory cells, in either the surface epithelium or the lamina propria. Three scores were identified according to the inflammatory cell density (1, rare and diffuse inflammation; 2, moderate inflammation; 3, severe inflammation).…”

Section: Methodsmentioning

confidence: 99%

CFTR and differentiation markers expression in non-CF and delta F 508 homozygous CF nasal epithelium.

Dupuit¹,

Kälin²,

Brézillon³

et al. 1995

J. Clin. Invest.

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Human nasal polyps from non-CF and AF 508 homozygous CF patients were used to compare the expression of CFTR and markers of epithelial differentiation, such as cytokeratins (CK) and desmoplakins (DP), at the transcriptional and translational levels. mRNA expression was assessed by semiquantitative RT/PCR kinetic assays while the expression and distribution of proteins were evaluated by immunofluorescence analysis. In parallel, for each nasal tissue specimen, the importance of surface epithelium remodeling and inflammation was estimated after histological observations. Our results show that the steady-state levels of CFTR, CK13, CK18, CK14, or DP 1 mRNA transcripts in AF 508 CF nasal polyps were not significantly different from those of non-CF tissues. A variability in the CFTR mRNA transcript level and in the pattern of CFTR immunolabeling has been observed between the different tissue samples. However, no relationship was found between the level of CFTR mRNA transcripts and the CFTR protein expression and distribution, either in the non-CF or in the CF group. The histological observations of non-CF and CF nasal polyp tissue indicated that the huge variations in the expression and distribution of the CFTR protein were associated with the variations in the degree of surface epithelium remodeling and inflammation in the lamina propria. A surface epithelium, showing a slight basal cell hyperplasia phenotype associated with diffuse inflammation, was mainly characterized by a CFTR protein distribution at the apex of ciliated cells in both non-CF and CF specimens. In contrast, in a remodeled surface epithelium associated with severe inflammation, CFTR protein presented either a diffuse distribution in the cytoplasm of ciliated cells, or was absent. These results suggest that abnormal expression and distribution of the CFTR protein in CF airways is not only caused by CFTR mutations. Airway surface epithelium remodeling and inflammation could play a critical role in the posttranscriptional and/ or the posttranslational regulation of the CFIR protein expression in non-CF and CF airways. (J. Clin. Invest. 1995Invest. . 95:1601Invest. -1611

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Immunohistochemical Localization of Cystic Fibrosis Transmembrane Conductance Regulator in Human Fetal Airway and Digestive Mucosa

Cited by 40 publications

References 8 publications

A mouse model for the delta F508 allele of cystic fibrosis.

A mouse model for the delta F508 allele of cystic fibrosis.

Cystic Fibrosis F508del Patients Have Apically Localized CFTR in a Reduced Number of Airway Cells

CFTR and differentiation markers expression in non-CF and delta F 508 homozygous CF nasal epithelium.

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