scholarly journals The Loss of Polysialic Acid Impairs the Contractile Phenotype of Peritubular Smooth Muscle Cells in the Postnatal Testis

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1347
Author(s):  
Nadim E. Hachem ◽  
Luisa Humpfle ◽  
Peter Simon ◽  
Miriam Kaese ◽  
Birgit Weinhold ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Smooth Muscle Actin ◽  
Polysialic Acid ◽  
Muscle Cells ◽  
Seminiferous Tubules ◽  
Vascular Smcs ◽  
Key Enzyme ◽  
Dependent Protein ◽  
Proliferating Cell

In the testis, the germinal epithelium of seminiferous tubules is surrounded by contractile peritubular cells, which are involved in sperm transport. Interestingly, in postnatal testis, polysialic acid (polySia), which is also an essential player for the development of the brain, was observed around the tubules. Western blotting revealed a massive decrease of polySia from postnatal day 1 towards puberty, together with a fundamental reduction of the net-like intertubular polySia. Using polysialyltransferase knockout mice, we investigated the consequences of the loss of polySia in the postnatal testis. Compared to postnatal wild-type animals, polySia knockouts showed slightly reduced smooth muscle actin (SMA) immunostaining of peritubular smooth muscle cells (SMCs), while calponin, marking more differentiated SMCs, dramatically decreased. In contrast, testicular SMA and calponin immunostaining remained unchanged in vascular SMCs in all genotypes. In addition, the cGMP-dependent protein kinase PKG I, a key enzyme of SMC relaxation, was nearly undetectable in the peritubular SMCs. Cell proliferation in the peritubular layer increased significantly in the knockouts, as shown by proliferating cell nuclear anti (PCNA) staining. Taken together, in postnatal testis, the absence of polySia resulted in an impaired differentiation of peritubular, but not vascular, SMCs to a more synthetic phenotype. Thus, polySia might influence the maintenance of a differentiated phenotype of non-vascular SMCs.

scholarly journals Peritubular Contractile Cells in Testis and Epididymis of the Dog, Canis lupus familiaris

2009 ◽  
Vol 78 (1) ◽  
pp. 3-11 ◽  
Author(s):  
Gunter F. Egger ◽  
Kirsti Witter
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Smooth Muscle Actin ◽  
Muscle Cells ◽  
Seminiferous Tubules ◽  
Intermediate Cell ◽  
Canis Lupus Familiaris ◽  
Efferent Ducts ◽  
Tubular System ◽  
Contractile Cells

Contractile cells surrounding the tubular system of the mammalian testis and epididymis are supposed to contribute to the initial transport of spermatozoa from the testis to epididymis. Testicular peritubular smooth muscle cells have been characterised in detail especially in rodents and humans. The aim of our study was to assess the distribution of peritubular contractile cells of the canine tubuli seminiferi, rete testis channels, ductuli efferentes, and ductus epididymidis by immunohistochemistry and transmission electron microscopy and to classify these cells with respect to their possible physiological function. The entire tubular system of the canine testis and epididymis is surrounded by contractile cells expressing smooth muscle actin, smooth muscle myosin and desmin, which are enveloped, at least partly, by a basal lamina. Some contractile cells of the tubuli seminiferi, rete channels, and efferent ducts and sometimes also single peritubular cells of the ductus epididymidis express vimentin. Contractile cells of seminiferous tubules and efferent ducts represent an intermediate cell type exhibiting characteristics of both smooth muscle cells (SMC) and myofibroblasts, those of rete channels stellate myofibroblasts, and those of the ductus epididymidis SMC. Differences in structure and arrangement of the contractile cells between seminiferous tubules, rete channels, efferent ducts, and ductus epididymidis suggest different functions. Myofibroblasts and contractile cells similar to them could be mainly responsible for the maintenance of an appropriate tissue turgor, whereas contraction of SMC of the ductus epididymidis might cause propulsion of spermatozoa by peristaltic waves.

Abstract 418: Co-Culture of Endothelial Cells, Smooth Muscle Cells and Monocytes in Collagen Scaffold: A Novel i n vitro Model of Atherosclerosis

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Martin Liu ◽  
Angelos Karagiannis ◽  
Matthew Sis ◽  
Srivatsan Kidambi ◽  
Yiannis Chatzizisis
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Type I Collagen ◽  
Smooth Muscle Actin ◽  
Muscle Cells ◽  
Type I ◽  
Collagen Gels ◽  
Human Coronary Artery

Objectives: To develop and validate a 3D in-vitro model of atherosclerosis that enables direct interaction between various cell types and/or extracellular matrix. Methods and Results: Type I collagen (0.75 mg/mL) was mixed with human artery smooth muscle cells (SMCs; 6x10 5 cells/mL), medium, and water. Human coronary artery endothelial cells (HCAECs; 10 5 /cm 2 ) were plated on top of the collagen gels and activated with oxidized low density lipoprotein cholesterol (LDL-C). Monocytes (THP-1 cells; 10 5 /cm 2 ) were then added on top of the HCAECs. Immunofluorescence showed the expression of VE-cadherin by HCAECs (A, B) and α-smooth muscle actin by SMCs (A). Green-labelled LDL-C particles were accumulated in the subendothelial space, as well as in the cytoplasm of HCAECs and SMCs (C). Activated monocytes were attached to HCAECs and found in the subendothelial area (G-I). Both HCAECs and SMCs released IL-1β, IL-6, IL-8, PDGF-BB, TGF-ß1, and VEGF. Scanning and transmission electron microscopy showed the HCAECs monolayer forming gap junctions and the SMCs (D-F) and transmigrating monocytes within the collagen matrix (G-I). Conclusions: In this work, we presented a novel, easily reproducible and functional in-vitro experimental model of atherosclerosis that has the potential to enable in-vitro sophisticated molecular and drug development studies.

scholarly journals Immunocytochemical characteristics of submucosal uterine myomas

2010 ◽  
Vol 67 (12) ◽  
pp. 977-982 ◽  
Author(s):  
Aleksandra Mladenovic-Mihailovic ◽  
Zorica Mladenovic-Bogdanovic ◽  
Predrag Mitrovic ◽  
Irena Tanaskovic ◽  
Slavica Usaj-Knezevic ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Connective Tissue ◽  
Smooth Muscle Cells ◽  
Smooth Muscle Actin ◽  
Muscle Cells ◽  
Synthetic Activity ◽  
Benign Tumors ◽  
Paraffin Sections ◽  
Weak Reaction ◽  
Synthetic Phenotype

Background/Aim. Myomas of the uterus, the most common benign tumors, have been studied for decades from the aspects of different basic and clinical disciplines. Despite this fact, their pathogenesis is still poorly understood. The aim of this study was to determine immunocytochemical characteristics of smooth muscle cells and connective tissue components of submucosal myomas of the uterus. Method. During the course of this study, 25 samples of submucosal myomas of the uterus were analyzed, all of them obtained during the surgery, after abdominal histerctomy by Aldridge. The samples were fixed in 4% formalin and embedded in paraffin. Sections of 5 ?m thickness were stained immunocytochemically using the DAKO LSAB+/HRP technique to identify ?- smooth muscle actin (?-SMA), vimentin, desmin, CD34, CD45, CD68 and PCNA (DAKO specification). Results. Our results suggest that submucosal myomas of the uterus are build-up of smooth muscle cells which are immunoreactive to ?-SMA and desmin, but also to a certain number of smooth muscle cells which are immunoreactive to ?-SMA and vimentin. Some of vimentin-immunoreactive cells also show an immunoreactivity of PCNA. In the build-up of connective stroma CD34-immunoreactive fibroblasts and neovascular formations are also present. By examining the distribution of CD45 antigen, at all the analyzed samples we observed a weak reaction. Conclusion. Submucosal myomas of the uterus are made-up of smooth muscle cells of the highly differentiated contractile phenotype (?-SMA- and desminimmunoreactivity), as well as smooth muscle cell of the synthetic phenotype which proliferate (?-SMA-, vimentin- and PCNA-immunoreactivity). In submucosal myoma of the uterus there is a significant presence of connective tissue as a result of synthetic activity of fibroblasts, which clearly differ in their immunocytochemical characteristics from smooth muscle cells of the synthetic phenotype.

Abstract 496: Extracellular Matrix Secretion by Vascular Smooth Muscle Cells: Role of MiR-195 and MiR-29b

2014 ◽  
Vol 34 (suppl_1) ◽  
Author(s):  
Anna Zampetaki ◽  
Xiaoke Yin ◽  
Ursula Mayr ◽  
Renata Gomes ◽  
Sarah Langley ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Abdominal Aortic Aneurysms ◽  
Aortic Aneurysms ◽  
Abdominal Aortic ◽  
Vascular Smcs

Rationale: Extracellular matrix (ECM) remodeling is a key function of vascular smooth muscle cells (SMCs). MicroRNAs (miRNAs), in particular the miR-29 family and miR-195, have been implicated in the control of ECM secretion. Objective: To perform a proteomics comparison of miRNA effects on ECM production by vascular SMCs. Methods and Results: Murine SMCs were transfected with miRNA mimics and antimiRs of miR-29b and miR-195, and their conditioned medium was analyzed by mass spectrometry. Both miRNAs targeted a cadre of ECM proteins, including proteoglycans, collagens, proteases, elastin and proteins associated with elastic microfibrils, albeit miR-29 showed a stronger effect. The proteomics findings were subsequently validated at the transcription level using quantitative polymerase chain reaction. Similar to miR-29, in vivo inhibition of miR-195 by intraperitoneal injection of cholesterol bound antagomiRs led to significant alterations of elastin expression in murine aortas. Since elastin degradation is a key event in aortic aneurysm formation, we investigated miR-195 expression in patients. In human aortic aneurysmal tissue, miR-195 expression was reduced compared to non-aneurysmal tissue. In plasma, a comparison between male patients with abdominal aortic aneurysms and controls matched for diabetes and hypertension returned a panel of five highly correlated miRNAs: miR-195, miR-125b, miR-148a, miR-20a and miR-340 showed significant inverse associations with the presence of abdominal aortic aneurysms and aortic diameter, with miR-195 dominating in terms of association strength. Conclusions: Using proteomic analysis, we compared the effect of miR-29 and miR-195 on ECM secretion by vascular SMCs and identified novel miRNA targets. Findings in patients support an important role for miR-195 in vascular remodeling as evidenced by reduced miR-195 expression in human aneurysmal tissue and an inverse correlation between plasma miR-195 levels and aortic diameter.

Conjugated Linoleic Acids Exert Similar Actions on Prostanoid Release from Aortic and Coronary Artery Smooth Muscle Cells

2006 ◽  
Vol 76 (5) ◽  
pp. 281-289 ◽  
Author(s):  
Ringseis ◽  
Gahler ◽  
Herter ◽  
Eder
Keyword(s):  
Smooth Muscle ◽  
Fatty Acid ◽  
Coronary Artery ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Biologically Active ◽  
Similar Degree ◽  
Conjugated Linoleic Acids ◽  
Vascular Smcs ◽  
Cla Isomers

Conjugated linoleic acids (CLAs) are biologically active lipid compounds exerting anti-atherogenic actions in vivo without exact knowledge about the underlying mechanisms. Recently, CLAs were shown to lower the release of vasoactive prostanoids from vascular smooth muscle cells (SMCs) which play a central role in atherosclerosis. Since SMCs from different vascular locations were shown to exert differential actions in response to a common stimulus, the present study aimed to explore potential differential effects of CLA isomers on the release of the prostanoids PGE2 and PGI2 from coronary artery and aortic SMCs. For this purpose, human aortic and coronary artery SMCs were incubated with 5 and 50 μmol/L of cis-9, trans-11 CLA and trans-10, cis-12 CLA for 24 hours and analyzed for fatty acid composition and the release of prostaglandins E2 and I2 (PGE2 and PGI2). Incubations were performed in the absence (basal conditions) and in the presence of 10 ng/mL of the cytokine tumor necrosis factor-α (TNFα) (cytokine-stimulated conditions). Fatty acid analysis revealed a similar degree of incorporation of CLA isomers and dose-dependent reduction of arachidonic acid in total cell lipids of both types of vascular SMCs following treatment with CLA. The release of PGE2 and PGI2 was dose-dependently inhibited by either CLA isomer from both types of vascular SMCs. The inhibitory potential of CLA isomers on the release of prostanoids was slightly different between basal and cytokine-stimulated conditions. In conclusion, the present findings suggest that the action of CLA isomers on the release of vasoactive prostanoids from vascular SMCs is largely independent of the vascular location; e.g., coronary arteries or systemic vasculature (aorta), but partially depends on the pathophysiological status of SMCs. The observed anti-inflammatory effect of CLAs may contribute to the anti-atherogenic actions of CLA.

scholarly journals Extracellular vesicle cross-talk between pulmonary artery smooth muscle cells and endothelium during excessive TGF-β signalling: implications for PAH vascular remodelling

2019 ◽  
Vol 17 (1) ◽  
Author(s):  
Fernando de la Cuesta ◽  
Ilaria Passalacqua ◽  
Julie Rodor ◽  
Raghu Bhushan ◽  
Laura Denby ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Pulmonary Artery ◽  
Smooth Muscle Cells ◽  
Smooth Muscle Actin ◽  
Muscle Cells ◽  
Protein Markers ◽  
Vascular Remodelling ◽  
Pulmonary Artery Smooth Muscle ◽  
Tgf Β1

Abstract Background Excessive TGF-β signalling has been shown to underlie pulmonary hypertension (PAH). Human pulmonary artery smooth muscle cells (HPASMCs) can release extracellular vesicles (EVs) but their contents and significance have not yet been studied. Here, we aimed to analyse the contents and biological relevance of HPASMC-EVs and their transport to human pulmonary arterial endothelial cells (HPAECs), as well as the potential alteration of these under pathological conditions. Methods We used low-input RNA-Seq to analyse the RNA cargoes sorted into released HPASMC-EVs under basal conditions. We additionally analysed the effects of excessive TGF-β signalling, using TGF-β1 and BMP4, in the transcriptome of HPASMCs and their EVs. We then, for the first time, optimised Cre-loxP technology for its use with primary cells in vitro, directly visualising HPASMC-to-HPAEC communication and protein markers on cells taking up EVs. Furthermore we could analyse alteration of this transport with excessive TGF-β signalling, as well as by other cytokines involved in PAH: IL-1β, TNF-α and VEGFA. Results We were able to detect transcripts from 2417 genes in HPASMC-EVs. Surprisingly, among the 759 enriched in HPASMC-EVs compared to their donor cells, we found Zeb1 and 2 TGF-β superfamily ligands, GDF11 and TGF-β3. Moreover, we identified 90 genes differentially expressed in EVs from cells treated with TGF-β1 compared to EVs in basal conditions, including a subset involved in actin and ECM remodelling, among which were bHLHE40 and palladin. Finally, using Cre-loxP technology we showed cell-to-cell transfer and translation of HPASMC-EV Cre mRNA from HPASMC to HPAECs, effectively evidencing communication via EVs. Furthermore, we found increased number of smooth-muscle actin positive cells on HPAECs that took up HPASMC-EVs. The uptake and translation of mRNA was also higher in activated HPAECs, when stimulated with TGF-β1 or IL-1β. Conclusions HPASMC-EVs are enriched in RNA transcripts that encode genes that could contribute to vascular remodelling and EndoMT during development and PAH, and TGF-β1 up-regulates some that could enhance this effects. These EVs are functionally transported, increasingly taken up by activated HPAECs and contribute to EndoMT, suggesting a potential effect of HPASMC-EVs in TGF-β signalling and other related processes during PAH development.

scholarly journals cGMP-dependent protein kinase I in vascular smooth muscle cells improves ischemic stroke outcome in mice

2019 ◽  
Vol 39 (12) ◽  
pp. 2379-2391
Author(s):  
Maria Shvedova ◽  
Maxim M Litvak ◽  
Jesse D Roberts ◽  
Dai Fukumura ◽  
Tomoaki Suzuki ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Ischemic Stroke ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Infarct Volume ◽  
Muscle Cells ◽  
Dependent Protein Kinase ◽  
Cgmp Dependent Protein Kinase ◽  
Dependent Protein

Recent works highlight the therapeutic potential of targeting cyclic guanosine monophosphate (cGMP)-dependent pathways in the context of brain ischemia/reperfusion injury (IRI). Although cGMP-dependent protein kinase I (cGKI) has emerged as a key mediator of the protective effects of nitric oxide (NO) and cGMP, the mechanisms by which cGKI attenuates IRI remain poorly understood. We used a novel, conditional cGKI knockout mouse model to study its role in cerebral IRI. We assessed neurological deficit, infarct volume, and cerebral perfusion in tamoxifen-inducible vascular smooth muscle cell-specific cGKI knockout mice and control animals. Stroke experiments revealed greater cerebral infarct volume in smooth muscle cell specific cGKI knockout mice (males: 96 ± 16 mm3; females: 93 ± 12 mm3, mean±SD) than in all control groups: wild type (males: 66 ± 19; females: 64 ± 14), cGKI control (males: 65 ± 18; females: 62 ± 14), cGKI control with tamoxifen (males: 70 ± 8; females: 68 ± 10). Our results identify, for the first time, a protective role of cGKI in vascular smooth muscle cells during ischemic stroke injury. Moreover, this protective effect of cGKI was found to be independent of gender and was mediated via improved reperfusion. These results suggest that cGKI in vascular smooth muscle cells should be targeted by therapies designed to protect brain tissue against ischemic stroke.

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