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Cell aggregation on agar as an indicator for cell-matrix adhesion: effects of opioids

Delphine Debruyne UGent, MARCUS MAREEL UGent, Barbara Vanhoecke UGent and Marc Bracke UGent (2009) IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL. 45(8). p.473-482
abstract
The slow aggregation assay is generally used to study the functionality of cell–cell adhesion complexes. Single cells are seeded on a semisolid agar substrate in a 96-well plate and the cells spontaneously aggregate. We used HEK FLAG-MOP cells that stably overexpress the mu opioid receptor and the mu-opioid-receptor-selective agonists DAMGO and morphine to study whether other factors than functionality of cell–cell adhesions complexes can contribute to changes in the pattern of slow aggregation on agar. HEK FLAG-MOP cells formed small compact aggregates. In the presence of DAMGO and morphine, larger and fewer aggregates were formed in comparison to the vehicle control. These aggregates were localized in the center of the agar surface, whereas in the vehicle control they were dispersed over the substrate. However, in suspension culture on a Gyrotory shaker, no stimulation of aggregation was observed by DAMGO and morphine, showing that opioids do not affect affinity. A dissociation experiment revealed that HEK FLAG-MOP aggregates formed in the absence or presence of opioids are resistant to de-adhesion. We demonstrated that the larger aggregates are neither the result of cell growth stimulation by DAMGO and morphine. Since manipulations of the substrate such as increasing the agar concentration or mixing agar with agarose induced the same changes in the pattern of slow aggregation as treatment with opioids, we suggest that cell–substrate adhesion may be involved in opioid-stimulated aggregation.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
Opioids, E-CADHERIN/CATENIN COMPLEX, Cell-substrate adhesion, Cell-cell adhesion, Aggregation, Agar, MAMMARY-CARCINOMA CELLS, INVASION, INTERNALIZATION
journal title
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
In Vitro Cell. Dev. Biol.-Anim.
volume
45
issue
8
pages
473 - 482
Web of Science type
Article
Web of Science id
000269320100009
JCR category
CELL BIOLOGY
JCR impact factor
0.791 (2009)
JCR rank
153/159 (2009)
JCR quartile
4 (2009)
ISSN
1071-2690
DOI
10.1007/s11626-009-9180-y
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
700343
handle
http://hdl.handle.net/1854/LU-700343
date created
2009-06-15 16:57:19
date last changed
2011-05-10 10:25:43
@article{700343,
  abstract     = {The slow aggregation assay is generally used to study the functionality of cell--cell adhesion complexes. Single cells are seeded on a semisolid agar substrate in a 96-well plate and the cells spontaneously aggregate. We used HEK FLAG-MOP cells that stably overexpress the mu opioid receptor and the mu-opioid-receptor-selective agonists DAMGO and morphine to study whether other factors than functionality of cell--cell adhesions complexes can contribute to changes in the pattern of slow aggregation on agar. HEK FLAG-MOP cells formed small compact aggregates. In the presence of DAMGO and morphine, larger and fewer aggregates were formed in comparison to the vehicle control. These aggregates were localized in the center of the agar surface, whereas in the vehicle control they were dispersed over the substrate. However, in suspension culture on a Gyrotory shaker, no stimulation of aggregation was observed by DAMGO and morphine, showing that opioids do not affect affinity. A dissociation experiment revealed that HEK FLAG-MOP aggregates formed in the absence or presence of opioids are resistant to de-adhesion. We demonstrated that the larger aggregates are neither the result of cell growth stimulation by DAMGO and morphine. Since manipulations of the substrate such as increasing the agar concentration or mixing agar with agarose induced the same changes in the pattern of slow aggregation as treatment with opioids, we suggest that cell--substrate adhesion may be involved in opioid-stimulated aggregation.},
  author       = {Debruyne, Delphine and MAREEL, MARCUS and Vanhoecke, Barbara and Bracke, Marc},
  issn         = {1071-2690},
  journal      = {IN VITRO CELLULAR \& DEVELOPMENTAL BIOLOGY-ANIMAL},
  keyword      = {Opioids,E-CADHERIN/CATENIN COMPLEX,Cell-substrate adhesion,Cell-cell adhesion,Aggregation,Agar,MAMMARY-CARCINOMA CELLS,INVASION,INTERNALIZATION},
  language     = {eng},
  number       = {8},
  pages        = {473--482},
  title        = {Cell aggregation on agar as an indicator for cell-matrix adhesion: effects of opioids},
  url          = {http://dx.doi.org/10.1007/s11626-009-9180-y},
  volume       = {45},
  year         = {2009},
}

Chicago
Debruyne, Delphine, Marcus Mareel, Barbara Vanhoecke, and Marc Bracke. 2009. “Cell Aggregation on Agar as an Indicator for Cell-matrix Adhesion: Effects of Opioids.” In Vitro Cellular & Developmental Biology-animal 45 (8): 473–482.
APA
Debruyne, Delphine, Mareel, M., Vanhoecke, B., & Bracke, M. (2009). Cell aggregation on agar as an indicator for cell-matrix adhesion: effects of opioids. IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, 45(8), 473–482.
Vancouver
1.
Debruyne D, Mareel M, Vanhoecke B, Bracke M. Cell aggregation on agar as an indicator for cell-matrix adhesion: effects of opioids. IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL. 2009;45(8):473–82.
MLA
Debruyne, Delphine, Marcus Mareel, Barbara Vanhoecke, et al. “Cell Aggregation on Agar as an Indicator for Cell-matrix Adhesion: Effects of Opioids.” IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL 45.8 (2009): 473–482. Print.