PMID- 8662885
OWN - NLM
STAT- MEDLINE
DCOM- 19960826
LR  - 20220310
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 271
IP  - 23
DP  - 1996 Jun 7
TI  - Dependence of epithelial intercellular junction biogenesis on 
      thapsigargin-sensitive intracellular calcium stores.
PG  - 13636-41
AB  - Perturbation of potentially regulatable endoplasmic reticulum (ER) calcium stores 
      with the Ca-ATPase inhibitor, thapsigargin (TG), perturbs the formation of 
      desmosomes and tight junctions during polarized epithelial cell biogenesis, 
      despite the development of cell contact. In a Madin-Darby canine kidney cell 
      model for intercellular junction assembly, TG treatment inhibited the development 
      of transepithelial electrical resistance (TER), a measure of tight junction 
      assembly, in a dose-dependent manner. The TG-induced inhibition of tight junction 
      assembly was paralleled by a defect in the sorting of the tight junction protein, 
      ZO-1. An even more dramatic delay in sorting of the desmosomal protein, 
      desmoplakin, was observed in the presence of TG. In addition, while both ZO-1 and 
      desmoplakin-I in control cells were shown to become associated with the Triton 
      X-100 insoluble cytoskeleton during intercellular junction assembly, prior 
      treatment with 100 nM TG diminished this biochemical stabilization into the 
      detergent-insoluble fraction, particularly in the case of ZO-1. Although 
      spectrofluorimetric measurements in fura-2 loaded Madin-Darby canine kidney cells 
      confirmed the occurrence of TG-mediated release of calcium from internal stores, 
      total cytosolic calcium during junction assembly remained similar to untreated 
      cells. Therefore, the presence of cytosolic calcium alone is not sufficient for 
      normal intercellular junction biogenesis if intracellular stores are perturbed by 
      TG. The results indicate the presence of calcium-sensitive intracellular 
      mechanisms involved in the sorting and cytoskeletal stabilization of both tight 
      junction and desmosomes and suggest a role for calcium-dependent signaling 
      pathways at an early (possibly common) step in polarized epithelial biogenesis.
FAU - Stuart, R O
AU  - Stuart RO
AD  - Renal Division, Department of Medicine, Brigham and Women's Hospital and Harvard 
      Medical School, Boston, Massachusetts 02115, USA.
FAU - Sun, A
AU  - Sun A
FAU - Bush, K T
AU  - Bush KT
FAU - Nigam, S K
AU  - Nigam SK
LA  - eng
GR  - 1 F32 DK08822-01A1/DK/NIDDK NIH HHS/United States
GR  - 5-T32-DK07527-09/DK/NIDDK NIH HHS/United States
GR  - R01DK44503/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Terpenes)
RN  - 67526-95-8 (Thapsigargin)
RN  - EC 7.2.2.10 (Calcium-Transporting ATPases)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - Calcium/*metabolism
MH  - Calcium-Transporting ATPases/antagonists & inhibitors
MH  - Cell Line
MH  - Dogs
MH  - Electric Impedance
MH  - Endoplasmic Reticulum/drug effects/metabolism
MH  - Enzyme Inhibitors/pharmacology
MH  - Epithelium/metabolism/ultrastructure
MH  - Intercellular Junctions/drug effects/*metabolism/ultrastructure
MH  - Intracellular Fluid/drug effects/metabolism
MH  - Macromolecular Substances
MH  - Terpenes/*pharmacology
MH  - Thapsigargin
EDAT- 1996/06/07 00:00
MHDA- 1996/06/07 00:01
CRDT- 1996/06/07 00:00
PHST- 1996/06/07 00:00 [pubmed]
PHST- 1996/06/07 00:01 [medline]
PHST- 1996/06/07 00:00 [entrez]
AID - S0021-9258(18)41370-1 [pii]
AID - 10.1074/jbc.271.23.13636 [doi]
PST - ppublish
SO  - J Biol Chem. 1996 Jun 7;271(23):13636-41. doi: 10.1074/jbc.271.23.13636.