Molecular cloning and expression of the chicken smooth muscle γ‐actin mRNA

Kovacs, Adrienne M.; Zimmer, Warren E.

doi:10.1002/cm.970240108

Cited by 12 publications

(2 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…36 Nevertheless, "combined " expression levels of smoothelin-A and-B were higher in bladder, consistent with smoothelin-A being expressed in visceral, not vascular, SMCs ( Figure 3C). Expression of telokin and smooth muscle ␥-actin, both visceral SMC markers, 37,38 was notably absent in SKP-derived SMCs but present in the adult rat bladder, providing further evidence that SKP-derived SMCs were not visceral in nature ( Figure 3D). Interestingly, nonmuscle myosin heavy chain B (NMMHC-B) (SMemb) was highly expressed in SKPderived SMCs and reached levels higher than in adult SMC-enriched tissues ( Figure 3C).…”

Section: Low Cell Density and Reduced Serum Concentrations Direct Ratmentioning

confidence: 59%

Directed Differentiation of Skin-Derived Precursors Into Functional Vascular Smooth Muscle Cells

Steinbach

El-Mounayri

DaCosta

et al. 2011

ATVB

View full text Add to dashboard Cite

Objective-The goal of this study was to characterize the factors and conditions required for smooth muscle cell (SMC)-directed differentiation of Sox2 ϩ multipotent rat and human skin-derived precursors (SKPs) and to define whether they represent a source of fully functional vascular SMCs for applications in vivo. Methods and Results-We found that rat SKPs can differentiate almost exclusively into SMCs by reducing serum concentrations to 0.5% to 2% and plating them at low density. Human SKPs derived from foreskin required the addition of transforming growth factor-␤1 or -␤3 to differentiate into SMCs, but they did so even in the absence of serum. SMC formation was confirmed by quantitative reverse transcription-polymerase chain reaction, immunocytochemistry, and fluorescence-activated cell sorting, with increased expression of smoothelin-B and little to no expression of telokin or smooth muscle ␥-actin, together indicating that SKPs differentiated into vascular rather than visceral SMCs. Rat and human SKP-derived SMCs were able to contract in vitro and also wrap around and support new capillary and larger blood vessel formation in angiogenesis assays in vivo. Conclusion-SKPs are Sox2ϩ progenitors that represent an attainable autologous source of stem cells that can be easily differentiated into functional vascular SMCs in defined serum-free conditions without reprogramming. SKPs represent a clinically viable cell source for potential therapeutic applications in neovascularization. (Arterioscler Thromb Vasc Biol. 2011;31:2938-2948.)

show abstract

Section: Low Cell Density and Reduced Serum Concentrations Direct Ratmentioning

confidence: 59%

Directed Differentiation of Skin-Derived Precursors Into Functional Vascular Smooth Muscle Cells

Steinbach

El-Mounayri

DaCosta

et al. 2011

ATVB

View full text Add to dashboard Cite

show abstract

“…7 Briefly, a chicken genomic library was constructed in EMBL-3 phage and was screened for SMGA clones using the 32 P-dCTP labeled full length cDNA, SMGA15-1, as a probe. 34 A unique XbaI restriction site was introduced at position +25 in exon 1 of the SMGA gene by PCR to facilitate cloning. The full length promoter was subcloned into the pCAT-basic vector (Promega, Madison, WI, USA) using the EcoRI and XbaI sites (Figure 1).…”

Section: Plasmidsmentioning

confidence: 99%

Cell-specific nuclear import of plasmid DNA

et al. 1999

View full text Add to dashboard Cite

One factor limiting the success of non-viral gene therapy smooth muscle cells, we have created a series of reporter vectors is the relative inability to target genes specifically plasmids that are expressed selectively in smooth muscle to a desired cell type. To address this limitation, we have cells. Moreover, when injected into the cytoplasm, plasbegun to develop cell-specific vectors whose specificity is mids containing portions of the SMGA promoter localize to at the level of the nuclear import of the plasmid DNA. We the nucleus of smooth muscle cells, but remain cytoplashave recently shown that nuclear import of plasmid DNA mic in fibroblasts and CV1 cells. In contrast, a similar plasis a sequence-specific event, requiring the SV40 enhancer, mid carrying the SV40 enhancer is transported into the a region known to bind to a number of general transcription nuclei of all cell types tested. Nuclear import of the SMGA factors (Dean DA, Exp Cell Res 1997; 230: 293). From promoter-containing plasmids could be achieved when the these studies we developed a model whereby transcription smooth muscle specific transcription factor SRF was factor(s) bind to the DNA in the cytoplasm to create a proexpressed in stably transfected CV1 cells, supporting our tein-DNA complex that can enter the nucleus using the model for the nuclear import of plasmids. Finally, these protein import machinery. Our model predicts that by using nuclear targeting sequences were also able to promote DNA elements containing binding sites for transcription facincreased gene expression in liposome-and polycationtors expressed in unique cell types, we should be able to transfected non-dividing cells in a cell-specific manner, create plasmids that target to the nucleus in a cell-specific similar to their nuclear import activity. These results promanner. Using the promoter from the smooth muscle vide proof of principle for the development of cell-specific gamma actin (SMGA) gene whose expression is limited to non-viral vectors for any desired cell type.

show abstract

The Developmentally Regulated Expression of Serum Response Factor Plays a Key Role in the Control of Smooth Muscle-Specific Genes

Browning

Culberson

Aragon

et al. 1998

Developmental Biology

View full text Add to dashboard Cite

Serum response factor (SRF) is a MADS box transcription factor that has been shown to be important in the regulation of a variety of muscle-specific genes. We have previously shown SRF to be a major component of multiple cis/trans interactions found along the smooth muscle gamma-actin (SMGA) promoter. In the studies reported here, we have further characterized the role of SRF in the regulation of the SMGA gene in the developing gizzard. EMSA analyses, using nuclear extracts derived from gizzards at various stages in development, showed that the SRF-containing complexes were not present early in gizzard smooth muscle development, but appeared as development progressed. We observed an increase in SRF protein and mRNA levels during gizzard development by Western and Northern blot analyses, with a large increase just preceding an increase in SMGA expression. Thus, changes in SRF DNA-binding activity were paralleled with increased SRF gene expression. Immunohistochemical analyses demonstrated a correspondence of SRF and SMGA expression in differentiating visceral smooth muscle cells (SMCs) during gizzard tissue development. This correspondence of SRF and SMGA expression was also observed in cultured smooth muscle mesenchyme induced to express differentiated gene products in vitro. In gene transfer experiments with SMGA promoter-luciferase reporter gene constructs we observed four- to fivefold stronger SMGA promoter activity in differentiated SMCs relative to replicating visceral smooth muscle cells. Further, we demonstrate the ability of a dominant negative SRF mutant protein to specifically inhibit transcription of the SMGA promoter in visceral smooth muscle, directly linking SRF with the control of SMGA gene expression. Taken together, these data suggest that SRF plays a prominent role in the developmental regulation of the SMGA gene.

show abstract

Molecular cloning and expression of the chicken smooth muscle γ‐actin mRNA

Cited by 12 publications

References 56 publications

Directed Differentiation of Skin-Derived Precursors Into Functional Vascular Smooth Muscle Cells

Directed Differentiation of Skin-Derived Precursors Into Functional Vascular Smooth Muscle Cells

Cell-specific nuclear import of plasmid DNA

The Developmentally Regulated Expression of Serum Response Factor Plays a Key Role in the Control of Smooth Muscle-Specific Genes

Contact Info

Product

Resources

About