Volume 126, Issue 1, Pages (January 2004)

Slides:



Advertisements
Similar presentations
Volume 72, Issue 2, Pages (July 2007)
Advertisements

Volume 15, Issue 3, Pages (March 2007)
Connective Tissue Growth Factor (CCN2) in Rat Pancreatic Stellate Cell Function: Integrin α5β1 as a Novel CCN2 Receptor  Runping Gao, David R. Brigstock 
Volume 6, Issue 5, Pages (November 2000)
Volume 73, Issue 11, Pages (June 2008)
Volume 136, Issue 3, Pages (March 2009)
Volume 128, Issue 1, Pages (January 2005)
Volume 125, Issue 5, Pages (November 2003)
Volume 127, Issue 2, Pages (August 2004)
by Mineo Iwata, Lynn Graf, Norihiro Awaya, and Beverly Torok-Storb
Volume 133, Issue 6, Pages (December 2007)
Volume 133, Issue 4, Pages (October 2007)
Volume 133, Issue 6, Pages (December 2007)
Base Treatment Corrects Defects Due to Misfolding of Mutant Cystic Fibrosis Transmembrane Conductance Regulator  Wan Namkung, Kyung Hwan Kim, Min Goo.
Volume 129, Issue 3, Pages (September 2005)
Volume 133, Issue 1, Pages (July 2007)
Volume 130, Issue 2, Pages (February 2006)
“Atypical p-ANCA” in IBD and hepatobiliary disorders react with a 50-kilodalton nuclear envelope protein of neutrophils and myeloid cell lines  Birgit.
Volume 136, Issue 5, Pages (May 2009)
Kwo–Yih Yeh, Mary Yeh, Jonathan Glass  Gastroenterology 
Volume 123, Issue 1, Pages (July 2002)
Volume 143, Issue 4, Pages e9 (October 2012)
Volume 129, Issue 5, Pages (November 2005)
Volume 127, Issue 2, Pages (August 2004)
Volume 126, Issue 1, Pages (January 2004)
Volume 127, Issue 3, Pages (September 2004)
Volume 126, Issue 7, Pages (June 2004)
Toll-like receptor 2 enhances ZO-1-associated intestinal epithelial barrier integrity via protein kinase C  Elke Cario, Guido Gerken, Daniel K. Podolsky 
Volume 134, Issue 1, Pages (January 2008)
Volume 142, Issue 2, Pages (February 2012)
John F. Öhd, Katarina Wikström, Anita Sjölander  Gastroenterology 
Volume 140, Issue 2, Pages (February 2011)
Volume 68, Issue 6, Pages (December 2005)
Duodenal expression of a putative stimulator of Fe transport and transferrin receptor in anemia and hemochromatosis  Donatella Barisani, Dario Conte 
Proteomic analysis of the slit diaphragm complex: CLIC5 is a protein critical for podocyte morphology and function  Brian A. Pierchala, Maura R. Muñoz,
Volume 136, Issue 4, Pages e3 (April 2009)
Volume 128, Issue 7, Pages (June 2005)
Volume 120, Issue 7, Pages (June 2001)
Volume 115, Issue 1, Pages (July 1998)
Volume 140, Issue 4, Pages e1 (April 2011)
Volume 137, Issue 3, Pages (September 2009)
Volume 136, Issue 2, Pages (February 2009)
Volume 123, Issue 3, Pages (September 2002)
Volume 133, Issue 4, Pages (October 2007)
Marie-Thérèse Leccia  Journal of Investigative Dermatology 
Volume 145, Issue 6, Pages e4 (December 2013)
Covering the Cover Gastroenterology
Volume 134, Issue 4, Pages (April 2008)
Volume 133, Issue 5, Pages (November 2007)
Activation of Epidermal Growth Factor Receptor/ERK Signaling Correlates with Suppressed Differentiation in Malignant Acanthosis Nigricans  Ingo Haase,
Differential Contribution of Dermal Resident and Bone Marrow–Derived Cells to Collagen Production during Wound Healing and Fibrogenesis in Mice  Reiichi.
The role of transforming growth factor beta-2, beta-3 in mediating apoptosis in the murine intestinal mucosa  Nicole Dünker, Kai Schmitt, Norbert Schuster,
Epidermal growth factor and transforming growth factor α down-regulate human gastric lipase gene expression  Eric Tremblay, Jean René Basque, Nathalie.
Volume 139, Issue 6, Pages (December 2010)
Volume 6, Issue 5, Pages (November 2000)
Volume 132, Issue 4, Pages (April 2007)
Volume 110, Issue 6, Pages (September 2002)
Noritaka Oyama, Keiji Iwatsuki, Yoshimi Homma, Fumio Kaneko 
Contribution of Src-FAK signaling to the induction of connective tissue growth factor in renal fibroblasts  A. Graness, I. Cicha, M. Goppelt-Struebe 
Connective Tissue Growth Factor (CCN2) in Rat Pancreatic Stellate Cell Function: Integrin α5β1 as a Novel CCN2 Receptor  Runping Gao, David R. Brigstock 
Volume 130, Issue 2, Pages (February 2006)
Temporal Regulation of Salmonella Virulence Effector Function by Proteasome- Dependent Protein Degradation  Tomoko Kubori, Jorge E. Galán  Cell  Volume.
Mitogen- and Stress-Activated Protein Kinase 2 and Cyclic AMP Response Element Binding Protein are Activated in Lesional Psoriatic Epidermis  Anne T.
Volume 139, Issue 2, Pages (August 2010)
Volume 76, Issue 1, Pages (July 2009)
Volume 126, Issue 1, Pages (January 2004)
This month in Gastroenterology
Katherine K. Rogers, Tzuu-Shuh Jou, Wei Guo, Joshua H. Lipschutz 
Gα12 and Gα13 Interact with Ser/Thr Protein Phosphatase Type 5 and Stimulate Its Phosphatase Activity  Yoshiaki Yamaguchi, Hironori Katoh, Kazutoshi Mori,
Matrix Metalloproteinase Inhibitor BB-3103 Unlike the Serine Proteinase Inhibitor Aprotinin Abrogates Epidermal Healing of Human Skin Wounds Ex Vivo1 
Presentation transcript:

Volume 126, Issue 1, Pages 32-41 (January 2004) The ΔF508 mutation results in loss of CFTR function and mature protein in native human colon  Marcus Mall, Silvia M. Kreda, April Mengos, Timothy J. Jensen, Stephanie Hirtz, Hans H. Seydewitz, James Yankaskas, Karl Kunzelmann, John R. Riordan, Richard C. Boucher  Gastroenterology  Volume 126, Issue 1, Pages 32-41 (January 2004) DOI: 10.1053/j.gastro.2003.10.049

Figure 1 Cholinergic and cAMP-dependent activation of ion transport in native normal and CF rectal epithelia. Experiments were performed in the presence of indomethacin (10 μmol/L, basolateral) and amiloride (10 μmol/L, luminal). Original recordings of the effects of carbachol (cch) and IBMX/forskolin on Vte and Rte in (A) normal and (B) CF (ΔF508/ΔF508) rectal biopsy specimens. Rte was determined from Vte deflections obtained by pulsed current injection. Time gaps between recordings were 20 minutes. (C) Summary of cAMP-induced (IBMX/forskolin) Isc in rectal biopsy specimens from wild-type (wt/wt), ΔF508 heterozygous (ΔF/wt), and ΔF508 homozygous CF (ΔF/ΔF) subjects. Individual data points represent the mean of measurements on 2–5 biopsy specimens per individual. ∗Significant difference comparing the effects of IBMX/forskolin in normal (wt/wt or ΔF/wt) and CF (ΔF/ΔF) rectal tissues (Kruskal-Wallis analysis of variance on ranks). (D) Summary of carbachol-induced Isc in the absence (−) and presence (+) of IBMX/forskolin. Closed circles, initial peak responses; open circles, the plateau for monophasic responses or the lumen-negative peak for biphasic responses in the absence of cAMP-dependent activation; closed diamonds, initial peak responses; open diamonds, the plateau for monophasic lumen-positive responses or the lumen-negative peak for biphasic responses in the presence of cAMP-dependent activation. #Significantly different from carbachol-induced plateau responses in rectal tissues from wild-type (wt/wt) and ΔF508 heterozygous (ΔF/wt) subjects in the absence of cAMP-dependent stimulation. ∗Significantly different from peak responses in wt/wt and ΔF/wt subjects in the presence of cAMP-dependent stimulation (Kruskal-Wallis analysis of variance on ranks). Solid line, median; dashed line, 25th and 75th percentiles, respectively. Gastroenterology 2004 126, 32-41DOI: (10.1053/j.gastro.2003.10.049)

Figure 2 CFTR protein detection in heterologous cells. (A) Specificity and sensitivity of monoclonal CFTR antibodies. (A, left panel) MTE18 cells lacking (m) or expressing (c) human CFTR were assayed by Western blot with identical concentrations (1 μg/mL) of monoclonal CFTR antibodies 769, 528, 596, and 217 (new) and 24-1 and MATG1061 (published). Mature (arrow) and immature (arrowhead) CFTR bands are indicated. (A, right panel) MTE18 cells lacking (mock) or expressing human CFTR grown on collagen-coated glass culture chambers were assayed independently for immunofluorescence with the indicated CFTR antibodies at identical concentrations (1 μg/mL). Nuclei were stained with 4′,6-diamidino-2-phenylindole. Cells were analyzed with a Leica 4D-TCS confocal microscope by xz scanning using 2 independent laser sources. (Original magnification 150×.) (B) BHK cells lacking (mock) or expressing either wild-type (wt) or ΔF508 (ΔF) CFTR were assayed by Western blot and immunofluorescence as previously described using monoclonal CFTR antibodies 769, 528, 596, and 217 as indicated. The mature CFTR band (arrow) is only present in BHK wt CFTR cells. The doublet evident in the ΔF508 BHK cells reflects the full-length core glycosylated protein and an N-terminally cleaved proteolysis product (upper and lower band, respectively). Molecular-weight bands are indicated in kilodaltons. Gastroenterology 2004 126, 32-41DOI: (10.1053/j.gastro.2003.10.049)

Figure 3 CFTR protein detection in freshly excised human colonic epithelia. Rectal biopsy specimens from normal (wt/wt; n = 3), ΔF508 homozygous (ΔF/ΔF; n = 3), and ΔF508 heterozygous (ΔF/wt; n = 3) individuals and the ascending colon of a normal individual (∗) were homogenized and immunoprecipitated with a polyclonal CFTR antibody. After sodium dodecyl sulfate/polyacrylamide gel electrophoresis (6% acrylamide), immunoblots were probed with a cocktail of CFTR monoclonal antibodies 596 and 217, which recognize different CFTR epitopes (see Patients and Methods). The fully glycosylated CFTR protein (arrow) was only observed in normal specimens. In contrast, CF samples exhibited only the immature CFTR protein (arrowhead). ΔF508 heterozygous specimens exhibited both fully processed and immature protein forms. Specimens from the rectum and ascending colon showed a similar pattern of CFTR protein expression. Molecular-weight bands are indicated in kilodaltons. Gastroenterology 2004 126, 32-41DOI: (10.1053/j.gastro.2003.10.049)

Figure 4 Morphology of freshly excised human rectal epithelia. (A) Schematic of rectal epithelium showing a transverse section of a deep crypt (left) and a single crypt (center). Colonocytes are shown in green and goblet cells in purple; nuclei are represented in blue. The actin cytoskeleton was labeled with Alexa 488 phalloidin to identify colonocytes and MUC2 antibody (color encode in purple) was used to label goblet cells (far right). l, lumen. (B) Rectal biopsy specimens from 10 normal (wt/wt) and 8 ΔF508 homozygous patients with CF (ΔF/ΔF) were immunostained with a MUC2 monoclonal antibody followed by a Texas Red-labeled secondary antibody and Alexa 488 phalloidin. Nuclei were stained with 4′,6-diamidino-2-phenylindole. Images show xy confocal planes scanned in a Leica SP2 AOBS confocal microscope in the differential interference contrast, 4′,6-diamidino-2-phenylindole, Texas Red (color encode in purple), and Alexa 488 channels using 3 independent laser sources. The antibody recognizes the MUC2 precursor protein and stains mainly the MUC2 associated with the endoplasmic reticulum of the goblet cells in both normal and CF crypts. (Bar = 20 μm.) Gastroenterology 2004 126, 32-41DOI: (10.1053/j.gastro.2003.10.049)

Figure 5 CFTR immunolocalization in freshly excised human rectal epithelia. Rectal biopsy specimens from 10 normal (wt/wt) and 8 age-matched ΔF508 homozygous individuals with CF (ΔF/ΔF) were immunostained independently with 3 CFTR antibodies (528, 769, and 525) and control immunoglobulin G at identical concentrations (1 μg/mL) followed by a Texas Red-labeled secondary antibody and Alexa 488 phalloidin (F-actin). Images are xy confocal planes recorded in the differential interference contrast, Texas Red, and Alexa 488 channels using 2 independent laser sources (488 and 568 nm) and identical scanning conditions. Representative results from (A) 1 normal and (B) 1 CF rectal tissue, stained with antibody 528, are shown. Control immunoglobulin G staining was negative in both normal and CF epithelia. Arrowheads indicate the position of the apical membrane of colonocytes. (Original magnification 100×.) Gastroenterology 2004 126, 32-41DOI: (10.1053/j.gastro.2003.10.049)