Selective isolation of rat aortic wall layers and their cell types in culture—Application to converting enzyme activity measurement

Battle, Thierry; Arnal, Jean‐François; Challah, Mireille; Michel, Jean‐Baptiste

doi:10.1016/0040-8166(94)90043-4

Cited by 97 publications

(63 citation statements)

References 40 publications

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“…Four hours later, the medium was changed and adherent cells were washed free of debris and non-attached cells. Smooth muscle and endothelial cells were obtained by a previously described method (13). The rat brain homogenate protein fraction was a gift from Dr. Ling Xiao.…”

Section: Methodsmentioning

confidence: 99%

Nuclear-delimited Angiotensin Receptor-mediated Signaling Regulates Cardiomyocyte Gene Expression

Tadevosyan

Maguy

Villeneuve

et al. 2010

Journal of Biological Chemistry

105

117

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Angiotensin-II (Ang-II) from extracardiac sources and intracardiac synthesis regulates cardiac homeostasis, with mitogenic and growth-promoting effects largely due to altered gene expression. Here, we assessed the possibility that angiotensin-1 (AT1R) or angiotensin-2 (AT2R) receptors on the nuclear envelope mediate effects on cardiomyocyte gene expression. Immunoblots of nucleus-enriched fractions from isolated cardiomyocytes indicated the presence of AT1R and AT2R proteins that copurified with the nuclear membrane marker nucleoporin-62 and histone-3, but not markers of plasma (calpactin-I), Golgi (GRP-78), or endoplasmic reticulum (GM130) membranes. Confocal microscopy revealed AT1R and AT2R proteins on nuclear membranes. Microinjected Ang-II preferentially bound to nuclear sites of isolated cardiomyocytes. AT1R and AT2R ligands enhanced de novo RNA synthesis in isolated cardiomyocyte nuclei incubated with [␣-

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Section: Methodsmentioning

confidence: 99%

Nuclear-delimited Angiotensin Receptor-mediated Signaling Regulates Cardiomyocyte Gene Expression

Tadevosyan

Maguy

Villeneuve

et al. 2010

Journal of Biological Chemistry

105

117

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“…20 This method provides pure cell preparations (Ͼ95% of endothelial cells), as shown by immunostaining with a specific antibody against rat endothelial cells. 21 Cells from passages 1 to 3 were used in these studies.…”

Section: Cell Culturementioning

confidence: 99%

Angiotensin II Stimulates Endothelial Vascular Cell Adhesion Molecule-1 via Nuclear Factor-κB Activation Induced by Intracellular Oxidative Stress

Pueyo

González

Nicoletti

et al. 2000

ATVB

493

306

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Abstract-The recruitment of monocytes via the endothelial expression of vascular cell adhesion molecule-1 (VCAM-1) is a key step in the formation of the initial lesion in atherosclerosis. Because angiotensin (Ang) II may be involved in this process, we investigated its role on the signaling cascade leading to VCAM-1 expression in endothelial cells. Ang II stimulates mRNA and protein expression of VCAM-1 in these cells via the AT 1 receptor. This effect was enhanced by N G -nitro-L-arginine methyl ester, a nitric oxide synthase inhibitor, and blocked by pyrrolidinedithiocarbamate, an antioxidant molecule. Ang II activated the redox-sensitive transcription factor nuclear factor-B and stimulated the degradation of both inhibitor of B (IB)␣ and IB␤ with different kinetics. The degradation of IBs induced by Ang II was not modified by incubation with exogenous superoxide dismutase and catalase, suggesting that this effect was not mediated by the extracellular production of O 2 Ϫ . In contrast, rotenone and antimycin, 2 inhibitors of the mitochondrial respiratory chain, inhibited the Ang II-induced IB degradation, showing that generation of reactive oxygen species in the mitochondria is involved on Ang II action. BXT-51702, a glutathione peroxidase mimic, inhibited the effect of Ang II, and aminotriazole, an inhibitor of catalase, enhanced it, suggesting a role for H 2 O 2 in IB degradation. This is confirmed by experiments showing that Ang II stimulates the intracellular production of H 2 O 2 in endothelial cells. These results demonstrate that Ang II induced an intracellular oxidative stress in endothelial cells, which stimulates IB degradation and nuclear factor-B activation. This activation enhances the expression of VCAM-1 and probably other genes involved in the early stages of atherosclerosis.

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“…The three cell types from aortae (ie, ECs from intima, SMCs from media, and fibroblasts from adventitia) of both Lou and BN inbred strains were isolated and cultured as described by Battle et al 39 The purity of the cultures was assessed using immunohistological and morphological criteria. ECs were stained for Von Willebrand factor and by RECA-1 (a monoclonal antibody specific for rat endothelial cells, kindly provided by Dr A.M. Duijvestijn, Maastricht, The Netherlands 40 ).…”

Section: Vascular Cell Phenotypesmentioning

confidence: 99%

“…Morphological criteria are described in detail elsewhere. 39 At passage 2, cells were cultured in six-well tissue culture plates. At the third day of confluency, cells were deprived of serum for 24 hours.…”

Section: Vascular Cell Phenotypesmentioning

confidence: 99%

Angiotensin I-Converting Enzyme Genotype Influences Arterial Response to Injury in Normotensive Rats

Challah

Villard

Philippe

et al. 1998

ATVB

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Abstract-Two normotensive strains of rat, the Lou and Brown Norway (BN) strains, have contrasting levels of plasma angiotensin-converting enzyme (ACE). To investigate the degree of genetic determination of ACE expression, a polymorphic marker of the ACE gene was analyzed in inbred rats of the two strains. The two inbred strains were shown to bear different alleles for a polymorphic marker at the ACE gene. The segregation of the alleles of this marker and the plasma ACE levels were studied in a group of F2 rats issued from a cross between Lou and BN rats. The degree of genetic determination of plasma ACE activity was estimated to be 94% in the F2 cohort. The ACE locus accounts for 74% of total plasma ACE variance. ACE activity and mRNA expression in lungs were also genetically determined. The difference observed in ACE mRNA accumulation in the lungs between the two strains was due to a difference in the transcriptional rate of the ACE gene, as shown in nuclear run-on experiments. No differences were observed in arterial blood pressure of homozygous F2 progeny. In these animals, ACE genotype did not interfere with the pressor or the depressor responses to ACE-dependent vasoactive peptides. There was a significant effect of strain on constitutive or inducible membrane or soluble ACE activity in primary cultures of vascular cells. Neointima formation in the carotid artery 14 days after balloon injury was also influenced by the genotype in F2 homozygous progeny, whereas the medial area was not. These results demonstrate that there is a close relationship between the genetically determined ACE expression and the inducibility of the ACE gene. The degree of genetic determination of ACE expression in inbred rat strains offers a unique opportunity to study the interaction between genetic and environmental determinants of ACE expression and its involvement in response to experimental cardiovascular and renal injury. (Arterioscler Thromb Vasc Biol. 1998;18:235-243.)

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Selective isolation of rat aortic wall layers and their cell types in culture—Application to converting enzyme activity measurement

Cited by 97 publications

References 40 publications

Nuclear-delimited Angiotensin Receptor-mediated Signaling Regulates Cardiomyocyte Gene Expression

Nuclear-delimited Angiotensin Receptor-mediated Signaling Regulates Cardiomyocyte Gene Expression

Angiotensin II Stimulates Endothelial Vascular Cell Adhesion Molecule-1 via Nuclear Factor-κB Activation Induced by Intracellular Oxidative Stress

Angiotensin I-Converting Enzyme Genotype Influences Arterial Response to Injury in Normotensive Rats

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