Differential Display and Cloning of Shear Stress-Responsive Messenger RNAs in Human Endothelial Cells

Ando, Jun; Tsuboi, Hiroko; Korenaga, Risa; Takahashi, K.; Kosaki, Keisuke; Isshiki, Masashi; Tojo, Tadashi; Takada, Yoshio; Kamiya, Akihide

doi:10.1006/bbrc.1996.1178

Cited by 47 publications

(29 citation statements)

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“…The microarray results confirmed findings reached by other methods on ET-1 (15), MCP-1 (16), and PAI-1 (7). Shear stress did not change NADH dehydrogenase; although Ando et al (17) found that a gene coding for a different subunit of this enzyme was down-regulated by shear stress. PGHS-2 mRNA was up-regulated at 6 h and returned to baseline by 24 h, which is consistent with our previous studies of PGHS-2 protein levels under similar shear conditions (18).…”

Section: Discussionmentioning

confidence: 90%

DNA microarray reveals changes in gene expression of shear stressed human umbilical vein endothelial cells

McCormick

Eskin

McIntire

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

343

218

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Using DNA microarray screening (GeneFilter 211, Research Genetics, Huntsville, AL) of mRNA from primary human umbilical vein endothelial cells (HUVEC), we identified 52 genes with significantly altered expression under shear stress [25 dynes͞cm 2 for 6 or 24 h (1 dyne ‫؍‬ 10 N), compared with matched stationary controls]; including several genes not heretofore recognized to be shear stress responsive. We examined mRNA expression of nine genes by Northern blot analysis, which confirmed the results obtained on DNA microarrays. Thirty-two genes were up-regulated (by more than 2-fold), the most enhanced being cytochromes P450 1A1 and 1B1, zinc finger protein EZF͞GKLF, glucocorticoid-induced leucine zipper protein, argininosuccinate synthase, and human prostaglandin transporter. Most dramatically decreased (by more than 2-fold) were connective tissue growth factor, endothelin-1, monocyte chemotactic protein-1, and spermidine͞spermine N1-acetyltransferase. The changes observed suggest several potential mechanisms for increased NO production under shear stress in endothelial cells. During the past 15 years, over 40 genes have been identified as being regulated by shear stress in endothelial cells (1-4). Shear stress responsive genes are involved in cell proliferation, differentiation, maintenance of vascular tone, thrombosis, cellmatrix and cell-cell adhesion, and modulation of the inflammatory͞immune system. The identification of such genes is important not only for developing a fundamental understanding of how endothelial cells work, but also for understanding and treating pathological conditions that are influenced by shear stress, such as thrombosis, restenosis, and atherosclerosis (5, 6).Most of the genes that have been shown to be regulated by shear stress were identified by using traditional techniques such as Northern blot analysis or reverse transcriptase PCR (7-9). The main limitation of these techniques is that only one gene or at best a handful of genes can be studied in one experiment. When multiple genes are studied by using traditional methods, the experiments usually require a reiteration of the detection procedure for each gene. Investigators must therefore be very selective in the genes they choose to study, necessitating a priori information linking the chosen genes to shear stress. Thus, these experiments generally tend to validate or disprove specific hypotheses and do not lead to the discovery of unexpected differentially expressed genes. However, DNA microarray technology allows researchers to study several thousands of genes at one time. In addition to identifying unexpected genes, this technology also has the power to lead to the development of new hypotheses concerning how cells respond to shear stress and identification of coregulated pathways responsive to the mechanical environment of the cell.We used DNA GeneFilter GF211 from Research Genetics (Huntsville, AL; ref. 10), which contains over 4,000 named human genes, to identify genes altered by shear stress in primary human umbilical vein en...

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Section: Discussionmentioning

confidence: 90%

DNA microarray reveals changes in gene expression of shear stressed human umbilical vein endothelial cells

McCormick

Eskin

McIntire

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

343

218

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“…LOX expression is immunohistochemically demonstrated in dermal fibroblasts and extracellular fibers in the chick and mouse embryos (Wakasaki and Ooshima, 1990;Hayashi et al, 2004) and human foreskin (Noblesse et al, 2004), although a temporospatial pattern of its expression still remains to be determined in the chick TMT skin. LOX expression is regulated variably in combinations of various effectors such as TGF-bs, interleukin-1b, prostagladin E 2 , and so on (Roy et al, 1996;Smith-Mungo and Kagan, 1997), but mechanical stress is also among effectors that activate LOX expression (Ando et al, 1996). In the onset of elastin deposition along MFs observed in the present study, mechanical stress can be significant, because MFs extending from ectodermal basal lamina penetrate fairly loose matrix at early stages (Hurle et al, 1989;Isokawa et al, 2004) but become buried in increasing amounts of orthogonally oriented collagen fibers as the fibrogenesis proceeds (Fig.…”

Section: Discussionmentioning

confidence: 99%

Late Deposition of Elastin to Vertical Microfibrillar Fibers in the Presumptive Dermis of the Chick Embryonic Tarsometatarsus

Yamazaki

Sejima

Yuguchi

et al. 2007

The Anatomical Record

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Fibrillin microfibrils are integral components of elastic fibers and serve as a scaffold for elastin deposition. However, microfibrillar fibers (MFs) are not necessarily committed to develop into so-called elastic fibers. In dermis, elastin-free oxytalan MFs originating from the dermoepidermal junction are continuous to elaunin-type MFs (with a small amount of elastin) in the deeper papillary dermis, whereas the reticular dermis contains elastic fibers, or MFs embedded largely in elastin. In this study, we have investigated temporospatial patterns of elastin deposition on the MFs in tarsometatarsal presumptive dermis. While the earliest expression of elastin was demonstrated immunohistochemically as early as embryonic day 4 (ED4) in the wall of cardiac outflow and pharyngeal arch arteries, its deposition in the tarsometatarsus was first detected at ED6 in the deeper mesenchyme and at ED13 in the subectodermal mesenchyme. In the latter tissue, MFs had been organized perpendicularly to the covering ectoderm by ED4, well before an overt accumulation of collagenous matrix. Elastin deposition was observed initially in a punctate manner at ED13 and afterward became continuous along MFs. However, a characteristic spaced array of subectodermal vertical MFs was disorganized by ED17. These findings suggest that elastin deposition in the subectodermal MFs is not deployed by continuous, orderly propagation from elastic fibers in the deeper mesenchyme but occurs de novo in multiple foci along vertical MFs. Moreover, the present chronology of elastin deposition indicates that subectodermal, elastin-free MFs function as a transient, but primary fibrous structure in the presumptive dermis before the accumulation of collagenous matrix.

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“…10 The passage of blood in the vessels generates hemodynamic forces, such as shear stress, and regulates the function of endothelial cells lining the intimal surface of the vasculature. 11 Shear stress stimulates the gene expression of CF6 12 as well as the synthesis and secretion of bioactive molecules such as prostacyclin, nitric oxide (NO), tissue plasminogen activator, and platelet-derived growth factor. [13][14][15] Recently, it was shown that the ␣ and ␤ subunits of ATP synthase are present on the surface in the vascular endothelial cells and that angiostatin binds to them to exert its effect, 16 -18 suggesting that ATP synthase on the plasma membrane may consist of full components.…”

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confidence: 99%

Mitochondrial Coupling Factor 6 Is Present on the Surface of Human Vascular Endothelial Cells and Is Released by Shear Stress

et al. 2001

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Background-We showed that mitochondrial coupling factor 6 (CF6), an endogenous inhibitor of prostacyclin synthesis, is present in the systemic circulation as a pressor substance in rats. We investigated the possibility of vascular endothelial cells as a source of circulating CF6. Methods and Results-We used 2 cultured endothelial cell lines, human umbilical vein endothelial cells (HUVECs) and ECV 304 cells (transformed HUVECs), for this study. Immunofluorescence microscopy of both ECV 304 and HUVECs confirmed the surface-associated immunoreactivity of anti-CF6 antibody on the plasma membrane. The concentration of CF6 in the medium increased gradually with time in both ECV 304 and HUVECs in static conditions. Exposure of ECV 304 and HUVECs to a fluid shear stress enhanced the release of CF6: In ECV 304, the concentration of CF6 in the medium (ng · well Ϫ1 · 6 hours

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Differential Display and Cloning of Shear Stress-Responsive Messenger RNAs in Human Endothelial Cells

Cited by 47 publications

References 0 publications

DNA microarray reveals changes in gene expression of shear stressed human umbilical vein endothelial cells

DNA microarray reveals changes in gene expression of shear stressed human umbilical vein endothelial cells

Late Deposition of Elastin to Vertical Microfibrillar Fibers in the Presumptive Dermis of the Chick Embryonic Tarsometatarsus

Mitochondrial Coupling Factor 6 Is Present on the Surface of Human Vascular Endothelial Cells and Is Released by Shear Stress

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