A variety of indirect data exist which, in aggregate, suggest that exocytosis can alter tight junction structure and/or permeability. These experiments are designed to assess the feasibility of utilizing a homogenous epithelia monolayer derived from a human intestinal cell line to study the effects of exocytosis on tight junction structure and permeability. To accomplish this goal we will utilize the 18N2 clone of the HT29 cell line which was derived from a human colonic carcinoma. HT 29-18N2 cells differentiate as goblet cells, grow as monolayers on coverslips, and respond to cholinergic stimulation with exocytosis of mucin granule content. We will first attempt to identify conditions under which HT2918N2 cells can be grown on permeable supports as confluent monolayers from which electrophysical and flux data can be obtained. Using physiological techniques we will then ascertain if stimulated exocytosis perturbs tight junction barrier function. Using electron microscopy and freeze fracture techniques we will define the effects which exocytosis exert on tight junction structure. Lastly we will define and inter-relate physiological and structural events which occur as monolayers recover from stimulated exocytosis. These studies may yield insights into how local events such as exocytosis influence tight junction barrier function. These studies may also define methods whereby electrophysiological studies can be performed on homogeneous, flat, easily manipulated monolayers composed entirely of one of the major subtypes of cells in the intestinal epithelium - the goblet cell.

Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Harvard University
Department
Type
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
Garcia-Castillo, Maria Daniela; Chinnapen, Daniel J-F; Te Welscher, Yvonne M et al. (2018) Mucosal absorption of therapeutic peptides by harnessing the endogenous sorting of glycosphingolipids. Elife 7:
Basu, Sankha S; Delaney, Mary L; Li, Ning et al. (2018) Acetobacter indonesiensis Pneumonia after Lung Transplantation. Emerg Infect Dis 24:598-599
Garber, John J; Mallick, Emily M; Scanlon, Karen M et al. (2018) Attaching-and-Effacing Pathogens Exploit Junction Regulatory Activities of N-WASP and SNX9 to Disrupt the Intestinal Barrier. Cell Mol Gastroenterol Hepatol 5:273-288
Maglic, Dejan; Schlegelmilch, Karin; Dost, Antonella Fm et al. (2018) YAP-TEAD signaling promotes basal cell carcinoma development via a c-JUN/AP1 axis. EMBO J 37:
Yien, Yvette Y; Shi, Jiahai; Chen, Caiyong et al. (2018) FAM210B is an erythropoietin target and regulates erythroid heme synthesis by controlling mitochondrial iron import and ferrochelatase activity. J Biol Chem 293:19797-19811
Cho, Jin A; Lee, Ann-Hwee; Platzer, Barbara et al. (2018) Retraction Notice to: The Unfolded Protein Response Element IRE1? Senses Bacterial Proteins Invading the ER to Activate RIG-I and Innate Immune Signaling. Cell Host Microbe 23:571
Long, Jonathan Z; Roche, Alexander M; Berdan, Charles A et al. (2018) Ablation of PM20D1 reveals N-acyl amino acid control of metabolism and nociception. Proc Natl Acad Sci U S A 115:E6937-E6945
Bader, Razan M; Jonas, Maureen M; Mitchell, Paul D et al. (2018) Controlled attenuation parameter: A measure of hepatic steatosis in patients with cystic fibrosis. J Cyst Fibros :
Ohsaki, Asa; Venturelli, Nicholas; Buccigrosso, Tess M et al. (2018) Maternal IgG immune complexes induce food allergen-specific tolerance in offspring. J Exp Med 215:91-113
Pacheco, Alline R; Lazarus, Jacob E; Sit, Brandon et al. (2018) CRISPR Screen Reveals that EHEC's T3SS and Shiga Toxin Rely on Shared Host Factors for Infection. MBio 9:

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