TY - JOUR
T1 - Apolipoprotein AI could be a significant determinant of epithelial integrity in rainbow trout gill cell cultures
T2 - A study in functional proteomics
AU - Smith, Richard W.
AU - Wood, Chris M.
AU - Cash, Phil
AU - Diao, Linda
AU - Pärt, Peter
N1 - Funding Information:
This study was financially supported by a European Commission contract between the JRC and Aberdeen University (contract number: 17033-2000-110F1ED ISP GB), and an NSERC Discovery Grant and a European Commission Visiting Fellowship awarded to CMW. CMW is supported by the Canada Research Chair Programme.
PY - 2005/6/15
Y1 - 2005/6/15
N2 - The freshwater fish gill forms a barrier against an external hypotonic environment. By culturing rainbow trout gill cells on permeable supports, as intact epithelia, this study investigates barrier property mechanisms. Under symmetrical conditions the apical and basolateral epithelial surfaces contact cell culture media. Replacing apical media with water, to generate asymmetrical conditions (i.e. the situation encountered by the freshwater gill), rapidly increases transepithelial resistance (TER). Proteomic analysis revealed that this is associated with enhanced expression of pre-apolipoprotein AI (pre-apoAI). To test the physiological relevance, gill cells were treated with a dose of 50 μg ml-1 human apolipoprotein (apoAI). This was found to elevate TER in those epithelia which displayed a lower TER prior to apoAI treatment. These results demonstrate the action of apoAI and provide evidence that the rainbow trout gill may be a site of apoAI synthesis. TER does not differentiate between the trans-cellular (via the cell membrane) and para-cellular (via intercellular tight junctions) pathways. However, despite the apoAI-induced changes in TER, para-cellular permeability (measured by polyethylene glycol efflux) remained unaltered suggesting apoAI specifically reduces trans-cellular permeability. This investigation combines proteomics with functional measurements to show how a proteome change may be associated with freshwater gill function.
AB - The freshwater fish gill forms a barrier against an external hypotonic environment. By culturing rainbow trout gill cells on permeable supports, as intact epithelia, this study investigates barrier property mechanisms. Under symmetrical conditions the apical and basolateral epithelial surfaces contact cell culture media. Replacing apical media with water, to generate asymmetrical conditions (i.e. the situation encountered by the freshwater gill), rapidly increases transepithelial resistance (TER). Proteomic analysis revealed that this is associated with enhanced expression of pre-apolipoprotein AI (pre-apoAI). To test the physiological relevance, gill cells were treated with a dose of 50 μg ml-1 human apolipoprotein (apoAI). This was found to elevate TER in those epithelia which displayed a lower TER prior to apoAI treatment. These results demonstrate the action of apoAI and provide evidence that the rainbow trout gill may be a site of apoAI synthesis. TER does not differentiate between the trans-cellular (via the cell membrane) and para-cellular (via intercellular tight junctions) pathways. However, despite the apoAI-induced changes in TER, para-cellular permeability (measured by polyethylene glycol efflux) remained unaltered suggesting apoAI specifically reduces trans-cellular permeability. This investigation combines proteomics with functional measurements to show how a proteome change may be associated with freshwater gill function.
KW - Apolipoprotein AI
KW - Proteome
KW - Trans-cellular flux
KW - Transepithelial resistance
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UR - http://www.scopus.com/inward/citedby.url?scp=17444381547&partnerID=8YFLogxK
U2 - 10.1016/j.bbapap.2005.02.009
DO - 10.1016/j.bbapap.2005.02.009
M3 - Article
C2 - 15848139
AN - SCOPUS:17444381547
VL - 1749
SP - 81
EP - 93
JO - BBA - Protein Structure
JF - BBA - Protein Structure
SN - 1570-9639
IS - 1
ER -