scholarly journals Combination of histochemical analyses and micro-MRI reveals regional changes of the murine cervix in preparation for labor

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Antara Chatterjee ◽  
Rojan Saghian ◽  
Anna Dorogin ◽  
Lindsay S. Cahill ◽  
John G. Sled ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Diffusion Tensor ◽  
Ex Vivo ◽  
Muscle Cells ◽  
Pregnant Mouse ◽  
Collagen Fibers ◽  
Muscular Layer ◽  
Tissue Organization ◽  
Picrosirius Red

AbstractThe cervix is responsible for maintaining pregnancy, and its timely remodeling is essential for the proper delivery of a baby. Cervical insufficiency, or “weakness”, may lead to preterm birth, which causes infant morbidities and mortalities worldwide. We used a mouse model of pregnancy and term labor, to examine the cervical structure by histology (Masson Trichome and Picrosirius Red staining), immunohistochemistry (Hyaluronic Acid Binding Protein/HABP), and ex-vivo MRI (T2-weighted and diffusion tensor imaging), focusing on two regions of the cervix (i.e., endocervix and ectocervix). Our results show that mouse endocervix has a higher proportion of smooth muscle cells and collagen fibers per area, with more compact tissue structure, than the ectocervix. With advanced gestation, endocervical changes, indicative of impending delivery, are manifested in fewer smooth muscle cells, expansion of the extracellular space, and lower presence of collagen fibers. MRI detected three distinctive zones in pregnant mouse endocervix: (1) inner collagenous layer, (2) middle circular muscular layer, and (3) outer longitudinal muscular layer. Diffusion MRI images detected changes in tissue organization as gestation progressed suggesting the potential application of this technique to non-invasively monitor cervical changes that precede the onset of labor in women at risk for preterm delivery.

scholarly journals Mechanical stretch is a highly selective regulator of gene expression in human bladder smooth muscle cells

2004 ◽  
Vol 20 (1) ◽  
pp. 36-44 ◽  
Author(s):  
Rosalyn M. Adam ◽  
Samuel H. Eaton ◽  
Carlos Estrada ◽  
Ashish Nimgaonkar ◽  
Shu-Ching Shih ◽  
...  
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Growth Factor ◽  
Smooth Muscle Cells ◽  
Ex Vivo ◽  
Muscle Cells ◽  
Mechanical Stimuli ◽  
Bladder Smooth Muscle ◽  
Human Bladder ◽  
Cox 2

Application of mechanical stimuli has been shown to alter gene expression in bladder smooth muscle cells (SMC). To date, only a limited number of “stretch-responsive” genes in this cell type have been reported. We employed oligonucleotide arrays to identify stretch-sensitive genes in primary culture human bladder SMC subjected to repetitive mechanical stimulation for 4 h. Differential gene expression between stretched and nonstretched cells was assessed using Significance Analysis of Microarrays (SAM). Expression of 20 out of 11,731 expressed genes (∼0.17%) was altered >2-fold following stretch, with 19 genes induced and one gene (FGF-9) repressed. Using real-time RT-PCR, we tested independently the responsiveness of 15 genes to stretch and to platelet-derived growth factor-BB (PDGF-BB), another hypertrophic stimulus for bladder SMC. In response to both stimuli, expression of 13 genes increased, 1 gene (FGF-9) decreased, and 1 gene was unchanged. Six transcripts (HB-EGF, BMP-2, COX-2, LIF, PAR-2, and FGF-9) were evaluated using an ex vivo rat model of bladder distension. HB-EGF, BMP-2, COX-2, LIF, and PAR-2 increased with bladder stretch ex vivo, whereas FGF-9 decreased, consistent with expression changes observed in vitro. In silico analysis of microarray data using the FIRED algorithm identified c-jun, AP-1, ATF-2, and neurofibromin-1 (NF-1) as potential transcriptional mediators of stretch signals. Furthermore, the promoters of 9 of 13 stretch-responsive genes contained AP-1 binding sites. These observations identify stretch as a highly selective regulator of gene expression in bladder SMC. Moreover, they suggest that mechanical and growth factor signals converge on common transcriptional regulators that include members of the AP-1 family.

scholarly journals Identification, Structural, and Functional Characterization of a New Early Gene (6A3-5, 7 kb): Implication in the Proliferation and Differentiation of Smooth Muscle Cells

2005 ◽  
Vol 2005 (3) ◽  
pp. 254-270 ◽  
Author(s):  
Kazem Zibara ◽  
Gwenaële Garin ◽  
John L. McGregor
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Ex Vivo ◽  
Functional Characterization ◽  
Muscle Cells ◽  
Protein Band ◽  
Early Gene ◽  
New Family ◽  
Transcription Profiles

Arterial smooth muscle cells (SMCs) play a major role in atherosclerosis and restenosis. Differential display was used to compare transcription profiles of synthetic SMCs to proliferating rat cultured SMC line. An isolated cDNA band (6A3-5) was shown by northern (7 kb) to be upregulated in the proliferating cell line. A rat tissue northern showed differential expression of this gene in different tissues. Using 5’ RACE and screening of a rat brain library, part of the cDNA was cloned and sequenced (5.4 kb). Sequence searches showed important similarities with a new family of transcription factors, bearing ARID motifs. A polyclonal antibody was raised and showed a protein band of 175 kd, which is localized intracellularly. We also showed that 6A3-5 is upregulated in dedifferentiated SMC (P9) in comparison to contractile SMC ex vivo (P0). This work describes cloning, structural, and functional characterization of a new early gene involved in SMC phenotype modulation.

scholarly journals A simple laboratory exercise with rat isolated esophagus and stomach fundus to reveal functional differences between striated and smooth muscle cells

2017 ◽  
Vol 41 (2) ◽  
pp. 291-297 ◽  
Author(s):  
Francisco José Batista-Lima ◽  
Kalinne Kelly Lima Gadelha ◽  
Daniel Maia Oliveira ◽  
Thiago Brasileiro Vasconcelos ◽  
Teresinha Silva Brito ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Student Learning ◽  
Ex Vivo ◽  
Electrical Field Stimulation ◽  
Muscle Cells ◽  
Physiological Solution ◽  
Force Frequency ◽  
Laboratory Activity ◽  
Laboratory Exercise

This study describes an undergraduate student laboratory activity using isolated preparations from rat gastrointestinal tissues that possess contractile profiles typically exhibited by striated and smooth muscle cells. While students are introduced to an ex vivo methodology, they can compare differences in trace experiments, twitch aspects, phasic and tonic properties, force-frequency relationships, and pharmacological responsiveness of esophageal (striated) and fundic (smooth muscle) segments. Muscle strips were subjected to electrical field stimulation (EFS) applied by platinum electrodes immersed in the physiological solution. The contractile profile of EFS responses varied between these two types of gut preparations. Atropine and tubocurarine revealed differential inhibitory influences in esophagus or fundus tissues; caffeine and procaine produced similar effects, i.e., potentiation and blockade of the EFS-induced contractile response in these tissues, respectively. Experimental results obtained during the activity helped the improvement of student learning about basic concepts previously discussed in theoretical lectures. To measure student learning with this laboratory exercise, a questionnaire was applied before and after the activity, and the number of expected correct answers, concerning the mechanisms of contraction in striated and smooth muscle, could be clearly evidenced.

scholarly journals Expression and Regulation of Glucocorticoid Receptor in Human Placental Villous Fibroblasts

Endocrinology ◽  
2005 ◽  
Vol 146 (11) ◽  
pp. 4619-4626 ◽  
Author(s):  
Men-Jean Lee ◽  
Zhen Wang ◽  
Herman Yee ◽  
Yuehong Ma ◽  
Nicole Swenson ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Human Placenta ◽  
Ex Vivo ◽  
Phosphorylation Site ◽  
Muscle Cells ◽  
Hormone Treatment ◽  
Cell Types ◽  
Mrna Levels ◽  
Stromal Fibroblasts

The human placenta is a glucocorticoid (GC)-responsive organ consisting of multiple cell types including smooth muscle cells, fibroblasts, and trophoblast that demonstrate changes in gene expression after hormone treatment. However, little is known about the relative expression or activity of the GC receptor (GR) among the various placental cell types. Normal term human placentas were examined by immunohistochemistry using either GR phosphorylation site-specific antibodies that are markers for various activation states of the GR or a GR antibody that recognizes the receptor independent of its phosphorylation state (total GR). We found strong total GR and phospho-GR immunoreactivity in stromal fibroblasts of terminal villi, as well as perivascular fibroblasts and vascular smooth muscle cells of the stem villi. Lower levels of both total GR and phospho-GR were found within cytotrophoblast cells relative to fibroblasts, whereas syncytiotrophoblast showed very little total GR or phospho-GR immunoreactivity. This pattern holds true for immunoblot analysis of extracts from cell fractions cultured ex vivo. In cultured placental fibroblasts, phosphorylation of GR increased upon short-term GC treatment, consistent with a role for GR phosphorylation in receptor transactivation. Total GR levels were reduced by nearly 90% after long-term hormone treatment; however, this down-regulation was independent of changes in GR mRNA levels. These findings demonstrate that GR levels in fibroblasts can be modulated by changes in hormone exposure. Such cell type-specific differences in GR protein expression and phosphorylation may provide the means of differentially regulating the GC response among the cells of the human placenta.

scholarly journals Stressor-Induced “Inflammaging” of Vascular Smooth Muscle Cells via Nlrp3-Mediated Pro-inflammatory Auto-Loop

2021 ◽  
Vol 8 ◽  
Author(s):  
Jaqueline Herrmann ◽  
Mengdi Xia ◽  
Manasa Reddy Gummi ◽  
Anna Greco ◽  
Annika Schacke ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Cardiovascular Mortality ◽  
Ex Vivo ◽  
Muscle Cells ◽  
Sterile Inflammation ◽  
Subsequent Reduction

Calcification of the vessel wall as one structural pathology of aged vessels is associated with high cardiovascular mortality of elderly patients. Aging is linked to chronic sterile inflammation and high burden of reactive oxygen species (ROS), leading to activation of pattern recognition receptors (PRRs) such as Nlrp3 in vascular cells. The current study investigates the role of PRR activation in the calcification of vascular smooth muscle cells (VSMCs). Therefore, in vitro cell culture of primary rat VSMCs and ex vivo aortic stimulations were used to analyze osteogenic, senescence and inflammatory markers via real-time PCR, in situ RNA hybridization, Western Blot, photometric assays and histological staining. Induction of ROS and DNA-damage by doxorubicin induces a shift of VSMC phenotype toward the expression of osteogenic, senescence and inflammatory proteins. Induction of calcification is dependent on Nlrp3 activity. Il-1β as a downstream target of Nlrp3 induces the synthetic, pro-calcifying VSMC phenotype. Inhibition of PRR with subsequent reduction of chronic inflammation might be an interesting target for reduction of calcification of VSMCs, with subsequent reduction of cardiovascular mortality of patients suffering from vessel stiffness.

Probing the Utility of Vascular Smooth Muscle Cells as a Target Cell for ex vivo Cardiovascular Gene Therapy

2000 ◽  
Vol 30 (6) ◽  
pp. 729
Author(s):  
Jonghoe Byun ◽  
Jeong Eun Huh ◽  
Eun A Jung ◽  
Sun Jin Park ◽  
Jin Ok Jeong ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Target Cell ◽  
Ex Vivo ◽  
Muscle Cells

scholarly journals Cellular Evidence of Telocytes as Novel Interstitial Cells within the Magnum of Chicken Oviduct

2017 ◽  
Vol 26 (1) ◽  
pp. 135-143 ◽  
Author(s):  
Ping Yang ◽  
Xudong Zhu ◽  
Lingling Wang ◽  
Nisar Ahmed ◽  
Yufei Huang ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Lamina Propria ◽  
Muscle Cells ◽  
Close Relation ◽  
Cell Types ◽  
Muscular Layer ◽  
3D Network ◽  
Close Relationship ◽  
Chicken Oviduct

Telocytes are a novel type of interstitial cell that has been identified in many organs of mammals, but there is little information available on these cells in avian species. This study shows the latest findings associated with telocytes in the muscular layer and lamina propria of the magnum of chicken oviduct analyzed by transmission electron microscopy. Telocytes are characterized by telopodes, which are thin and long prolongations, and a small amount of cytoplasm rich with mitochondria. Spindle- or triangular-shaped telocytes were detected at various locations in the magnum. In the muscular layer, telocytes have direct connection with smooth muscle cells. The cell body of telocytes along with their long telopodes mainly exists in the interstitial space between the smooth muscle bundles, whereas large numbers of short telopodes are scattered in between the smooth muscle cells. In the lamina propria, extremely long telopodes are twisting around each other and are usually collagen embedded. Both in the lamina propria and muscular layer, telocytes have a close relationship with other cell types, such as immune cells and blood vessels. Telopodes appear with dichotomous branching alternating between the podom and podomer, forming a 3D network structure with complex homo- and heterocellular junctions. In addition, a distinctive size of the vesicles is visible around the telopodes and may be released from telopodes because of the close relation between the vesicle and telopode. All characteristics of telocytes in the magnum indicate that telocytes may play a potential, but important, role in the pathogenesis of oviduct diseases.

scholarly journals Ultrasonically targeted delivery into endothelial and smooth muscle cells in ex vivo arteries

2007 ◽  
Vol 118 (3) ◽  
pp. 285-293 ◽  
Author(s):  
Daniel M. Hallow ◽  
Anuj D. Mahajan ◽  
Mark R. Prausnitz
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Targeted Delivery ◽  
Ex Vivo ◽  
Muscle Cells

scholarly journals SAMD1 Distribution Patterns in Mouse Atherosclerosis Models Suggest Roles in LDL Retention, Antigen Presentation, and Cell Phenotype Modulation

2021 ◽  
Author(s):  
Bruce Campbell ◽  
Patricia Bourassa ◽  
Robert Aiello
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Foam Cell ◽  
Ex Vivo ◽  
Muscle Cells ◽  
Distribution Patterns ◽  
Lesion Size ◽  
Cell Phenotype ◽  
Lesion Growth

The theory that lesions formed by retention of circulating LDL can then progress to complicated atherosclerotic lesions has been a subject of debate, as has the mechanism of retention. In earlier work, we identified SAMD1, a protein expressed by intimal smooth muscle cells in human lesions that appears to irreversibly bind apoB-Lps in extracellular matrix near the lumen. We hypothesized this binding could contribute to the formation of lesions in mice, and that inhibiting binding could reduce lesion growth. In mouse models of atherosclerosis, we found that SAMD1 binds LDL; that SAMD1/apoB complex is ingested by intimal cells; and that recognizable epitopes of the SAMD1/apoB complex survive some degree of catabolism in foam cell. These data appear to support the SAMD1/LDL retention hypothesis of lesion growth. Despite apparently irreversible binding of human LDL to full-length human SAMD1, efficient anti-SAMD1-antibody inhibitors were created. In vivo lesion targeting of inhibitors was demonstrated by MRI, ultrasound, and ex vivo microscopy. However, only non-statistically significant reductions in spontaneous lesion size in apoE-/- mice were seen after 12 weeks of treatment with PEG-fab inhibitors of SAMD1/LDL binding. In contrast, inhibitors substantially reduced LDL retention in carotid injury lesions in apoE-/- and LDLR-/- mice 7 days after injury. The most obvious difference between injury lesions and early spontaneous lesions is the presence of numerous SMCs and associated ECM in the injury lesions. Thus, SAMD1 may be involved in retention of apoB-Lps in mouse lesions, but not until smooth muscle cells have entered the intima. In addition, SAMD1 is seen throughout arteries in changing patterns that suggest broader and more complicated roles in atherosclerosis.

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