Estrogen decreases biglycan mRNA expression in resistance blood vessels

2003 ◽  
Vol 285 (4) ◽  
pp. R754-R761 ◽  
Author(s):  
Manoj C. Rodrigo ◽  
Douglas S. Martin ◽  
Kathleen M. Eyster
Keyword(s):  
Gene Expression ◽  
Microarray Data ◽  
Dna Microarrays ◽  
Mesenteric Arteries ◽  
Ovariectomized Rats ◽  
Rt Pcr ◽  
New Gene ◽  
The Media ◽  
Estrogen Effects ◽  
Estrogenic Regulation

This study was designed to identify new gene targets of estrogen in the mesenteric arteries and to determine whether the soy phytoestrogens could mimic estrogen effects. Ovariectomized rats were treated with estradiol, genistein, or daidzein for 4 days. The mesenteric arteries were harvested, total RNA was extracted, mRNA was reverse transcribed in the presence of [33P]dCTP, and the labeled probes were hybridized with DNA microarrays. Analysis of the microarray data identified biglycan as a target of estrogenic regulation. Semiquantitative RT-PCR was used to confirm and quantitate the decrease in biglycan gene expression in response to estrogen (-37%), genistein (-15%), and daidzein (-10%). Treatment with the pure estrogen receptor antagonist ICI-182,780 reversed the inhibition of biglycan gene expression. The decrease in biglycan gene expression in response to estrogens was paralleled with a decrease in biglycan protein expression. Biglycan protein was localized to the media of the mesenteric arteries by immunohistochemistry. Collectively, these data suggest that biglycan is a vascular protein regulated at the genomic level by estrogens.

Vascular gene expression in mice overexpressing human endothelin-1 targeted to the endothelium

2011 ◽  
Vol 43 (3) ◽  
pp. 148-160 ◽  
Author(s):  
Stefania M. C. Simeone ◽  
Melissa W. Li ◽  
Pierre Paradis ◽  
Ernesto L. Schiffrin
Keyword(s):  
Gene Expression ◽  
Vascular Injury ◽  
Expression Profiling ◽  
Dna Microarrays ◽  
Vascular Diseases ◽  
Vascular Wall ◽  
Mesenteric Arteries ◽  
Blood Pressure Elevation ◽  
Genome Wide ◽  
Lipid Metabolism Genes

Endothelin (ET)-1 plays an important pathophysiological role in several vascular diseases including hypertension and atherosclerosis. Transgenic mice overexpressing human preproET-1 selectively in the endothelium (eET-1) exhibit vascular injury in the absence of blood pressure elevation. ET-1 overexpression may induce vascular injury by inducing changes in gene expression. To understand mechanisms whereby ET-1 induces vascular damage, vascular gene expression profiling was performed using DNA microarrays. RNA from mesenteric arteries of male and female young (6–7 wk) and mature (6–8 mo) eET-1 and wild-type (WT) mice was isolated, and changes in gene expression were determined by genome-wide expression profiling using Illumina microarray and FlexArray software. Data were analyzed using a relaxed and a stringent statistical approach. The gene lists were compared and analyzed as well with Ingenuity Pathway Analysis. The most common change was an increase in the expression of lipid metabolism genes. Four of these genes were validated by qPCR, cyp51, dgat2, and scd1 genes in young and elovl6 in both young and mature male mice, supporting a role of ET-1 in atherosclerosis. To test the hypothesis that ET-1 participates in mechanisms leading to atherosclerosis, we crossed eET-1 with atherosclerosis-prone apoE−/− mice to determine whether ET-1 overexpression exacerbates high-fat diet (HFD)-induced atherosclerosis using oil red O staining of descending thoracic aorta. HFD increased lipid plaques by 3-, 27-, and 86-fold in eET-1, apoE−/−, and crossed mice, respectively, vs. WT. This suggests that increased endothelial ET-1 expression results in early changes in gene expression in the vascular wall that enhance lipid biosynthesis and accelerate progression of atherosclerosis.

scholarly journals 90 Establishment and characterization of Day 30 equine chorionic girdle and allantochorion cell lines

2020 ◽  
Vol 32 (2) ◽  
pp. 171
Author(s):  
S. Salman ◽  
A. Asghar ◽  
C. Magee ◽  
Q. Winger ◽  
G. Bouma ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Chorionic Gonadotropin ◽  
Vector System ◽  
Stable Cell Line ◽  
Rt Pcr ◽  
Study Gene Expression ◽  
Lentiviral Infection ◽  
The Media ◽  
Equine Chorionic Gonadotropin

Establishing cell lines is a good model for experimental applications to study molecular mechanisms and cell-specific gene expression. Equids have a diffuse epitheliochorial placenta, where the invasive trophoblast is represented by the chorionic girdle (CG) and the noninvasive trophoblast by the allantochorion (AC). Embryonic CG cells are unique to horses and have a crucial role in equine chorionic gonadotropin (eCG) production and maintenance of pregnancy during the first trimester. This study had three objectives: (1) establishing a stable cell line from Day 30 CG cells and AC using lentivirus encoding hTERT; (2) characterisation of Day 30 CG cells and AC cell morphology and expression of eCG α (eCGA) and β (eCGB) subunits, major histocompatibility complex class II (MHCII), and Kisspeptin receptor (KISS1R) in CG and AC cells; (3) investigating eCG protein production invitro from Day 30 CG and AC cells. Three mares (n=3) were used to collect Day 30 conceptuses by non-surgical uterine lavage on Day 30 of pregnancy. All 3 conceptuses were dissected for CG and AC cells then cultured invitro to confluency in cell culture plates. Second-generation lentiviral particles were generated using a three-vector system including transfer vector pLV-hTERT-IRES-hygro, and human telomerase reverse transcriptase (hTERT) lentivirus was utilised to establish stable hygromycin-resistant equine embryonic cell lines. Reverse-transcription PCR (RT-PCR) was used to study gene expression in cells and radioimmunoassay was used to investigate protein presence in the media. We established a hygromycin-resistant Day 30 CG and AC cell lines that express eCGA, eCGB, and hTERT and confirmed using RT-PCR yielding the predicted bands. The cell lines were maintained for 16 passages (7±2 days/passage), 10 of which were cultured after the lentiviral infection steps. Also, we characterised CG cells as fast-growing, large, binucleated, and epithelioid, and AC cells as rapid-growing showing smaller, squamous, mononucleate, epithelioid, and elongated fibroblastic cells. The RT-PCR results showed eCGA and eCGB subunits are expressed by both Day 30 CG and AC cells, but MHCII and KISS1R genes were not expressed in either of cells. Moreover, radioimmunoassay results showed that Day 30 CG cells did produce eCG protein (35.42ngmL−1) invitro earlier than what previous literature has shown. However, Day 30 AC cells did not produce eCG protein (0.042ngmL−1) invitro, and both CG and AC cell lines stopped secreting eCG in the media after the lentiviral infection. To conclude, establishing stable and hygromycin-resistant cell lines from Day 30 equine CG and AC cells using lentivirus encoding pLV-hTERT-IRES-hygro is attainable. Also, equine chorionic gonadotropin eCG protein is produced invitro as early as Day 30 from CG cells.

scholarly journals Differential expression of VEGFA, TIE2, and ANG2 but not ADAMTS1 in rat mesenteric microvascular arteries and veins

2009 ◽  
pp. 193-202
Author(s):  
N Mecha Disassa ◽  
B Styp-Rekowska ◽  
B Hinz ◽  
L Da Silva-Azevedo ◽  
AR Pries ◽  
...  
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Shear Stress ◽  
Wall Stress ◽  
Mesenteric Arteries ◽  
Rt Pcr ◽  
Hemodynamic Forces ◽  
Differential Gene ◽  
Structural Adaptations ◽  
Arteries And Veins

Microvessels respond to metabolic stimuli (e.g. pO2) and hemodynamic forces (e.g. shear stress and wall stress) with structural adaptations including angiogenesis, remodeling and pruning. These responses could be mediated by differential gene expression in endothelial and smooth muscle cells. Therefore, rat mesenteric arteries and veins were excised by microsurgery, and mRNA expression of four angioadaptation-related genes was quantified by real time duplex RT-PCR in equal amounts of total RNA, correlated to two different house keeping genes (ß-actin, GAPDH). The results show higher expression of VEGFA, TIE2, and ANG2 in arteries than in veins, but equal expression of ADAMTS1. Higher availability of VEGFA mRNA in endothelial cells of arteries shown here could contribute to the maintenance of mechanically stressed blood vessels and counteract pressureinduced vasoconstriction.

1650: Novel Gene Expression Markers in Renal Tumors Using Rt-PCR on Fixed Tissues Useful in Differentiating Histologic Subtypes

2004 ◽  
Vol 171 (4S) ◽  
pp. 436-436
Author(s):  
John A. Petros ◽  
Audry N. Schuetz ◽  
Andrew N. Young ◽  
Q. Yin Goen ◽  
So Dug Lim ◽  
...  
Keyword(s):  
Gene Expression ◽  
Renal Tumors ◽  
Rt Pcr ◽  
Novel Gene ◽  
Histologic Subtypes

Gene expression of collagen, fibronectin and elastin on a collagen and a collagen-fabric matrix measured by realtime RT-PCR

2004 ◽  
Vol 52 (S 1) ◽  
Author(s):  
LO Schilling ◽  
N Davies ◽  
R Autschbach ◽  
P Zilla
Keyword(s):  
Gene Expression ◽  
Rt Pcr
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