Amyloid angiopathy of the human brain: immunohistochemical studies using markers for components of extracellular matrix, smooth muscle actin and endothelial cells

1998 ◽  
Vol 96 (6) ◽  
pp. 558-563 ◽  
Author(s):  
W. W. Zhang ◽  
Hariklia Lempessi ◽  
Yngve Olsson
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Smooth Muscle ◽  
Human Brain ◽  
Smooth Muscle Actin ◽  
Amyloid Angiopathy ◽  
Muscle Actin ◽  
Immunohistochemical Studies

Osteopontin expression in spontaneously developed neointima in fowl (Gallus gallus)

2000 ◽  
Vol 203 (2) ◽  
pp. 273-282
Author(s):  
R.J. Kuykindoll ◽  
H. Nishimura ◽  
D.B. Thomason ◽  
S.K. Nishimoto
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Abdominal Aorta ◽  
Smooth Muscle Actin ◽  
Polyacrylamide Gel Electrophoresis ◽  
Balloon Catheter ◽  
Protein Band ◽  
Muscle Actin ◽  
Smooth Muscle Tissue ◽  
Aortic Smooth Muscle

Fowl show spontaneous elevation of blood pressure and neointimal plaque formation in the abdominal aorta at young ages. A similar neointima can be induced by a balloon-catheter-induced endothelium injury to the fowl aorta. Both spontaneously developed and injury-induced vascular lesions exhibit subendothelial hyperplasia consisting of neointimal cells with a synthetic phenotype and abundant extracellular matrix. The role of the extracellular matrix in the formation of neointima is not known. In this study, we investigated whether osteopontin, an adhesive glycoprotein present in the extracellular matrix, is expressed in aortic smooth muscle tissue of the fowl abdominal aorta, in spontaneously developed neointimal plaques and in the aortic smooth muscle underlying neointimal plaques. Crude protein extracted from isolated aortic smooth muscle tissues and neointimal plaques was fractionated by SDS-polyacrylamide gel electrophoresis and analyzed by immunoblotting with rabbit anti-fowl osteopontin (provided by Dr L. C. Gerstenfeld, Boston University) or anti-α smooth muscle actin antibodies. The anti-fowl osteopontin antibody predominantly recognized a 66–70 kDa protein band in neointimal plaques that co-migrated with the osteopontin phosphoprotein from chick bone. In contrast, intact aortic smooth muscle and the smooth muscle underlying neointimal plaques equally expressed three proteins (66–70 kDa, approximately 50 kDa and approximately 43 kDa) recognized by the anti-osteopontin antibody. Anti-α smooth muscle actin antibody recognized a 43 kDa protein band, and the expression of α smooth muscle actin was higher in aortic smooth muscle than in neointimal plaques. Osteopontin mRNA expression was examined using reverse transcription-polymerase chain reaction (RT-PCR) of total RNA from vascular tissues with specific primers constructed on the basis of the reported fowl osteopontin nucleotide sequence. The PCR products from intact aortic smooth muscle and neointimal plaques correspond to the product from recombinant plasmid cDNA (a gift from Dr L. C. Gerstenfeld) transcribed in vitro. These results suggest that osteopontin is synthesized in intact aortic smooth muscle and neointimal plaques in fowl and that unmetabolized approximately 66 kDa osteopontin protein is a predominant form in the neointima, indicating that osteopontin protein may be actively synthesized in the neointima.

scholarly journals Host-Derived Pericytes and Sca-1+ Cells Predominate in the MART-1− Stroma Fraction of Experimentally Induced Melanoma

2011 ◽  
Vol 59 (12) ◽  
pp. 1060-1075 ◽  
Author(s):  
J. Humberto Treviño-Villarreal ◽  
Douglas A. Cotanche ◽  
Rosalinda Sepúlveda ◽  
Magda E. Bortoni ◽  
Otto Manneberg ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Endothelial Cells ◽  
Smooth Muscle ◽  
Blood Vessel ◽  
Smooth Muscle Actin ◽  
Cell Types ◽  
Muscle Actin ◽  
Cellular Composition ◽  
Tumor Capsule ◽  
Form Blood Vessel

Identification of cell types in tumor-associated stroma that are involved in the development of melanoma is hampered by their heterogeneity. The authors used flow cytometry and immunohistochemistry to demonstrate that anti–MART-1 antibodies can discriminate between melanoma and stroma cells. They investigated the cellular composition of the MART-1−, non-hematopoietic melanoma-associated stroma, finding it consisted mainly of Sca-1+ and CD146+ cells. These cell types were also observed in the skin and muscle adjacent to developing melanomas. The Sca-1+ cell population was observed distributed in the epidermis, hair follicle bulges, and tumor capsule. The CD146+ population was found distributed within the tumor, mainly associated with blood vessels in a perivascular location. In addition to a perivascular distribution, CD146+ cells expressed α-smooth muscle actin, lacked expression of endothelial markers CD31 and CD34, and were therefore identified as pericytes. Pericytes were found to be associated with CD31+ endothelial cells; however, some pericytes were also observed associated with CD31−, MART-1+ B16 melanoma cells that appeared to form blood vessel structures. Furthermore, the authors observed extensive nuclear expression of HIF-1α in melanoma and stroma cells, suggesting hypoxia is an important factor associated with the melanoma microenvironment and vascularization. The results suggest that pericytes and Sca-1+ stroma cells are important contributors to melanoma development.

Endothelial cells genetically selected from differentiating mouse embryonic stem cells incorporate at sites of neovascularization in vivo

2002 ◽  
Vol 115 (10) ◽  
pp. 2075-2085 ◽  
Author(s):  
Sandrine Marchetti ◽  
Clotilde Gimond ◽  
Kristiina Iljin ◽  
Christine Bourcier ◽  
Kari Alitalo ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Endothelial Cell ◽  
Smooth Muscle Cell ◽  
Muscle Cell ◽  
Es Cells ◽  
Smooth Muscle Actin ◽  
Embryonic Stem ◽  
Muscle Actin ◽  
Puromycin Selection

Large scale purification of endothelial cells is of great interest as it could improve tissue transplantation, reperfusion of ischemic tissues and treatment of pathologies in which an endothelial cell dysfunction exists. In this study, we describe a novel genetic approach that selects for endothelial cells from differentiating embryonic stem (ES) cells. Our strategy is based on the establishment of ES-cell clones that carry an integrated puromycin resistance gene under the control of a vascular endothelium-specific promoter, tie-1. Using EGFP as a reporter gene, we first confirmed the endothelial specificity of the tie-1 promoter in the embryoid body model and in cells differentiated in 2D cultures. Subsequently, tie-1-EGFP ES cells were used as recipients for the tie-1-driven puror transgene. The resulting stable clones were expanded and differentiated for seven days in the presence of VEGF before puromycin selection. As expected, puromycin-resistant cells were positive for EGFP and also expressed several endothelial markers, including CD31, CD34,VEGFR-1, VEGFR-2, Tie-1, VE-cadherin and ICAM-2. Release from the puromycin selection resulted in the appearance of α-smooth muscle actin-positive cells. Such cells became more numerous when the population was cultured on laminin-1 or in the presence of TGF-β1, two known inducers of smooth muscle cell differentiation. The hypothesis that endothelial cells or their progenitors may differentiate towards a smooth muscle cell phenotype was further supported by the presence of cells expressing both CD31 andα-smooth muscle actin markers. Finally, we show that purified endothelial cells can incorporate into the neovasculature of transplanted tumors in nude mice. Taken together, these results suggest that application of endothelial lineage selection to differentiating ES cells may become a useful approach for future pro-angiogenic and endothelial cell replacement therapies.

scholarly journals Extracellular Matrix Scaffold for Cardiac Repair

Circulation ◽  
2005 ◽  
Vol 112 (9_supplement) ◽  
Author(s):  
Keith A. Robinson ◽  
Jinshen Li ◽  
Megumi Mathison ◽  
Alka Redkar ◽  
Jianhua Cui ◽  
...  
Keyword(s):  
Heart Failure ◽  
Flow Cytometry ◽  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Urinary Bladder ◽  
Smooth Muscle Actin ◽  
Left Ventricular ◽  
Foreign Body Response ◽  
Muscle Actin ◽  
Extracellular Matrix Scaffold

Background— Heart failure remains a significant problem. Tissue-engineered cardiac patches offer potential to treat severe heart failure. We studied an extracellular matrix scaffold for repairing the infarcted left ventricle. Methods and Results— Pigs (n=42) underwent left ventricular (LV) infarction. At 6 to 8 weeks, either 4-layer multilaminate urinary bladder-derived extracellular matrix or expanded polytetrafluoroethlyene (ePTFE) was implanted as full-thickness LV wall patch replacement. At 1-week, 1-month, or 3-month intervals, pigs were terminated. After macroscopic examination, samples of tissue were prepared for histology, immunocytochemistry, and analysis of cell proportions by flow cytometry. One-week and 1-month patches were intact with thrombus and inflammation; at 1 month, there was also tissue with spindle-shaped cells in proteoglycan-rich and collagenous matrix. More α-smooth muscle actin-positive cells were present in urinary bladder matrix (UBM) than in ePTFE (22.2±3.3% versus 8.4±2.7%; P =0.04). At 3 months, UBM was bioresorbed, and a collagen-rich vascularized tissue with numerous myofibroblasts was present. Isolated regions of α-sarcomeric actin-positive, intensely α-smooth muscle actin-immunopositive, and striated cells were observed. ePTFE at 3 months had foreign-body response with necrosis and calcification. Flow cytometry showed similarities of cells from UBM to normal myocardium, whereas ePTFE had limited cardiomyocyte markers. Conclusions— Appearance of a fibrocellular tissue that included contractile cells accompanied biodegradation of UBM when implanted as an LV-free wall infarction patch. UBM appears superior to synthetic material for cardiac patching and trends toward myocardial replacement at 3 months.

scholarly journals Cardiac microvascular endothelial cells express α-smooth muscle actin and show low NOS III activity

1999 ◽  
Vol 276 (5) ◽  
pp. H1755-H1768 ◽  
Author(s):  
Hiroshi Ando ◽  
Thomas Kubin ◽  
Wolfgang Schaper ◽  
Jutta Schaper
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Actin ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Microvascular Endothelial Cells ◽  
Low Grade ◽  
Muscle Actin ◽  
Microvascular Endothelial

We established a culture system of porcine coronary microvascular endothelial cells (MVEC) with high cellular yield and purity >98%. Endothelial origin was confirmed by immunostaining, immunoblotting and fluorescence-activated cell sorter (FACS) analysis using low-density lipoprotein uptake, CD31, von Willebrand factor, and the lectin Dolichos biflorus agglutinin. MVEC were positive for α-smooth muscle actin in culture and in myocardium, as confirmed by FACS. Of the primary MVEC, ∼30% expressed nitric oxide synthase (NOS) III in numbers decreasing from the first passage (6 ± 1%) to the second passage (4 ± 1%; P < 0.001 vs. primary isolates), whereas ∼100% of aortic endothelial cells (AEC) expressed NOS III. In AEC, NOS III activity (pmol citrulline ⋅ mg protein−1 ⋅ min−1) was 80 ± 10 and was nearly abolished in the absence of calcium (5 ± 1, P < 0.001). In primary MVEC, however, NOS III activity in the presence and absence of calcium was 20 ± 4 and 25 ± 5, respectively. We conclude that cardiac MVEC, in contrast to AEC, contain α-smooth muscle actin, show low-grade NOS III activity, and provide a suitable in vitro system for the study of endothelial pathophysiology.

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