Select de novo gene and protein expression during renal epithelial cell culture in rotating wall vessels is shear stress dependent.
The rotating wall vessel has gained popularity as a clinical cell culture tool to produce hormonal implants. It is desirable to understand the mechanisms by which the rotating wall vessel induces genetic changes, if we are to prolong the useful life of implants. During rotating wall vessel culture gravity is balanced by equal and opposite hydrodynamic forces including shear stress. The current study provides the first evidence that shear stress response elements, which modulate gene expression in endothelial cells, are also active in epithelial cells. Rotating wall culture of renal cells changes expression of select gene products including the giant glycoprotein scavenger receptors cubulin and megalin, the structural microvillar protein villin, and classic shear stress response genes ICAM, VCAM and MnSOD. Using a putative endothelial cell shear stress response element binding site as a decoy, we demonstrate the role of this sequence in the regulation of selected genes in epithelial cells. However, many of the changes observed in the rotating wall vessel are independent of this response element. It remains to define other genetic response elements modulated during rotating wall vessel culture, including the role of hemodynamics characterized by 3-dimensionality, low shear and turbulence, and cospatial relation of dissimilar cell types.
Nephrology Section, Department of Medicine and Tulane Environmental Astrobiology Center, Tulane/Xavier Center for Bioenvironmental Research, 1430 Tulane Avenue, New Orleans, LA 70112, USA.
J H
JH
Kaysen
W C
WC
Campbell
R R
RR
Majewski
F O
FO
Goda
G L
GL
Navar
F C
FC
Lewis
T J
TJ
Goodwin
T G
TG
Hammond
D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000235:genetics; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002454:Cell Differentiation; D011992:Endosomes / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D006112:Gravitation; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics; D026541:Heymann Nephritis Antigenic Complex; D006801:Humans; D007672:Kidney Cortex / Q000166:cytology* / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D019736:Prostheses and Implants; D051381:Rats; D017207:Rats, Sprague-Dawley; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012399:Rotation; D013314:Stress, Mechanical; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000235:genetics
D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D006360:Heat-Shock Proteins; D026541:Heymann Nephritis Antigenic Complex; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D016376:Oligonucleotides, Antisense; D011956:Receptors, Cell Surface; C029486:intrinsic factor-cobalamin receptor; C026615:villin; D013482:Superoxide Dismutase