scholarly journals IL-6 Protein Expression is Increased in Fast Glycolytic Muscle Fibers Over Time in the SOD1G93A Mouse model.

2021 ◽  
Author(s):  
Laura Moreno Martínez ◽  
Ana Cristina Calvo ◽  
Leticia Moreno-García ◽  
Gabriel García-Salamero ◽  
Christian Lunetta ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Patterns ◽  
Muscle Fibers ◽  
Main Role ◽  
Inflammatory Biomarker ◽  
Gene And Protein Expression ◽  
Edl Muscle ◽  
Als Patients ◽  
Lateral Sclerosis ◽  
Over Time

Abstract The deregulation of the inflammatory cytokine interleukin (IL)-6 has been associated to a variety of neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). The aim of this work was to analyze the variation of IL-6 levels in blood and damaged tissues during the course of the disease. We studied IL-6 protein expression in spinal cord, extensor digital longus (EDL) muscle and soleus (SOL) muscle of the SOD1G93A animal model at four stages of the disease by western blot. Concurrently, we analyzed IL-6 gene and protein expression in blood of ALS patients, healthy subjects and patients with other neuropathies through RTqPCR and ELISA. The results revealed different expression patterns depending on both the tissue analyzed and the stage of the disease, showing increasing IL-6 levels in EDL muscle over time. Moreover, lower IL-6 levels in blood were found in ALS patients. The decreased levels of IL-6 in blood from ALS patients could suggest that IL-6 might not be the main pro-inflammatory biomarker in the last stages in whole blood. In contrast, IL-6 may play a main role in fast glycolytic muscle fibers associated with muscle atrophy, suggesting that modulation of IL-6 in this tissue could be a potential target for anti-inflammatory therapies in ALS.

scholarly journals Tissue ischemia time affects gene and protein expression patterns within minutes following surgical tumor excision

BioTechniques ◽  
2004 ◽  
Vol 36 (6) ◽  
pp. 1030-1037 ◽  
Author(s):  
Annika Spruessel ◽  
Garnet Steimann ◽  
Mira Jung ◽  
Sung A. Lee ◽  
Theresa Carr ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Patterns ◽  
Tumor Excision ◽  
Ischemia Time ◽  
Tissue Ischemia ◽  
Gene And Protein Expression

scholarly journals Cell signaling pathways in human mutant PAX6 corneal cells: an in vitro model for aniridia-related keratopathy

2021 ◽  
Author(s):  
Marta Słoniecka ◽  
André Vicente ◽  
Berit Byström ◽  
Fátima Pedrosa Domellöf
Keyword(s):  
Protein Expression ◽  
Signaling Pathways ◽  
Cell Fate ◽  
Expression Patterns ◽  
Pax6 Gene ◽  
Cas9 Nuclease ◽  
Extracellular Matrix Components ◽  
Gene And Protein Expression ◽  
Human Keratocytes

ABSTRACTPURPOSETo establish an in vitro model of aniridia-related keratopathy (ARK) using CRISPR/Cas9 engineered human keratocytes with mutations in the PAX6 gene, and to study the Notch Homolog 1, Translocation-Associated (Notch1), sonic hedgehog (SHH), mammalian target of rapamycin (mTOR), and Wnt/β-catenin signaling pathways in the PAX6 mutant keratocytes.METHODSPrimary human keratocytes were isolated from healthy corneas. Keratocytes were transduced with Cas9 lentiviral particles in order to create cells stably expressing Cas9 nuclease. Lentiviral particles carrying PAX6 sgRNA were transduced into the Cas9 keratocytes creating mutants. Analysis of signaling pathways was assessed by RT-qPCR for gene expression and western blot for protein expression.RESULTSHuman keratocytes stably expressing Cas9 nuclease were created. Keratocytes carrying PAX6 gene mutation were successfully generated. PAX6 mutant keratocytes showed modified expression patterns of extracellular matrix components such as collagens and fibrotic markers. Analysis of the Notch1, SHH, mTOR, and Wnt/β-catenin signaling pathways in the PAX6 mutant keratocytes revealed altered gene and protein expression of the key players involved in these pathways.CONCLUSIONSA properly functioning PAX6 gene in keratocytes is crucial for the regulation of signaling pathways important for cell fate determination, proliferation, and inflammation. Pax6 mutation in the in vitro settings leads to changes in these pathways which resemble those found in corneas of patients with ARK.

Growth surface-induced gene and protein expression patterns in Caco-2 cells

2008 ◽  
Vol 4 (6) ◽  
pp. 1819-1826 ◽  
Author(s):  
Claudia Piana ◽  
Stefan Toegel ◽  
Iris Guell ◽  
Stefan Gerbes ◽  
Helmut Viernstein ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Patterns ◽  
Growth Surface ◽  
Gene And Protein Expression

Differential gene and protein expression in abluminal sprouting and intraluminal splitting forms of angiogenesis

2006 ◽  
Vol 110 (5) ◽  
pp. 587-595 ◽  
Author(s):  
James L. Williams ◽  
Alexander Weichert ◽  
Andreas Zakrzewicz ◽  
Luis Da Silva-Azevedo ◽  
Axel R. Pries ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Patterns ◽  
Gene Array ◽  
Matrix Metalloproteinase 2 ◽  
Adult Skeletal Muscle ◽  
Longitudinal Splitting ◽  
Sprouting Angiogenesis ◽  
Reverse Transcription Pcr ◽  
Gene And Protein Expression ◽  
Angiopoietin 2

In adult skeletal muscle, abluminal sprouting or longitudinal splitting of capillaries can be initiated separately by muscle overload and elevated microcirculation shear stress respectively. In the present study, gene and protein expression patterns associated with the different forms of angiogenesis were examined using a targeted gene array (Superarray), validated by quantitative RT (reverse transcription)-PCR and immunoblots. Sprouting angiogenesis induced large changes in expression levels in genes associated with extracellular matrix remodelling, such as MMP-2 (matrix metalloproteinase-2), TIMP (tissue inhibitor of metalloproteinases), SPARC (secreted protein, acidic and rich in cysteine) and thrombospondin. Changes in neuropilin, midkine and restin levels, which may underpin changes in endothelial morphology, were seen during splitting angiogenesis. Up-regulation of VEGF (vascular endothelial growth factor), Flk-1, angiopoietin-2 and PECAM-1 (platelet/endothelial cell adhesion molecule-1) was seen in both forms of angiogenesis, representing a common angiogenic response of endothelial cells. In conclusion, the present study demonstrates that general angiogenic signals from growth factors can be influenced by the local microenvironment resulting in differing forms of capillary growth to produce a co-ordinated expansion of the vascular bed.

Relaxin and Milk-Borne Lactocrine-Acting Factors Affect Morphoregulatory Gene and Protein Expression Patterns in the Neonatal Porcine Uterus.

2009 ◽  
Vol 81 (Suppl_1) ◽  
pp. 257-257
Author(s):  
Joseph C. Chen ◽  
Amy-Lynn Frankshun ◽  
Anne A. Wiley ◽  
Dori J. Miller ◽  
Kristene A. Welch ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Patterns ◽  
Gene And Protein Expression

scholarly journals Discordance between GLP-1R gene and protein expression in mouse pancreatic islet cells

2020 ◽  
Vol 295 (33) ◽  
pp. 11529-11541 ◽  
Author(s):  
Sarah M. Gray ◽  
Yurong Xin ◽  
Elizabeth C. Ross ◽  
Bryanna M. Chazotte ◽  
Megan E. Capozzi ◽  
...  
Keyword(s):  
Protein Expression ◽  
Islet Cells ◽  
Expression Patterns ◽  
Metabolic Stress ◽  
Β Cells ◽  
Β Cell ◽  
Protein Levels ◽  
Gene And Protein Expression ◽  
Glucagon Like Peptide 1 ◽  
Mouse Pancreatic Islet

The insulinotropic actions of glucagon-like peptide 1 receptor (GLP-1R) in β-cells have made it a useful target to manage type 2 diabetes. Metabolic stress reduces β-cell sensitivity to GLP-1, yet the underlying mechanisms are unknown. We hypothesized that Glp1r expression is heterogeneous among β-cells and that metabolic stress decreases the number of GLP-1R–positive β-cells. Here, analyses of publicly available single-cell RNA-Seq sequencing (scRNASeq) data from mouse and human β-cells indicated that significant populations of β-cells do not express the Glp1r gene, supporting heterogeneous GLP-1R expression. To check these results, we used complementary approaches employing FACS coupled with quantitative RT-PCR, a validated GLP-1R antibody, and flow cytometry to quantify GLP-1R promoter activity, gene expression, and protein expression in mouse α-, β-, and δ-cells. Experiments with Glp1r reporter mice and a validated GLP-1R antibody indicated that >90% of the β-cells are GLP-1R positive, contradicting the findings with the scRNASeq data. α-cells did not express Glp1r mRNA and δ-cells expressed Glp1r mRNA but not protein. We also examined the expression patterns of GLP-1R in mouse models of metabolic stress. Multiparous female mice had significantly decreased β-cell Glp1r expression, but no reduction in GLP-1R protein levels or GLP-1R–mediated insulin secretion. These findings suggest caution in interpreting the results of scRNASeq for low-abundance transcripts such as the incretin receptors and indicate that GLP-1R is widely expressed in β-cells, absent in α-cells, and expressed at the mRNA, but not protein, level in δ-cells.

scholarly journals Chromatin run-on sequencing analysis finds that ECM remodeling plays an important role in canine hemangiosarcoma pathogenesis

2020 ◽  
Author(s):  
Chinatsu Mukai(New Corresponding Author) ◽  
Eunju Choi ◽  
Kelly L. Sams ◽  
Elena Zu Klampen ◽  
Lynne Anguish ◽  
...  
Keyword(s):  
Protein Expression ◽  
Differentially Expressed Genes ◽  
Tumor Tissue ◽  
Expression Patterns ◽  
Unmet Need ◽  
Splenic Tissue ◽  
Differentially Expressed ◽  
Sequencing Analysis ◽  
Ecm Remodeling ◽  
Gene And Protein Expression

Abstract Background Canine visceral hemangiosarcoma (HSA) is a highly aggressive cancer of endothelial origin that closely resembles human visceral angiosarcoma, both clinically and histopathologically. Currently there is an unmet need for new diagnostics and therapies for both forms of this disease. The goal of this study was to utilize ChRO-seq and immunohistochemistry (IHC) to identify gene and protein expression signatures that may be important drivers of HSA progression. Methods Chromatin run-on sequencing (ChRO-seq) was performed on tissue isolated from 17 HSA samples and 4 normal splenic samples. Computational analysis was then used to identify differentially expressed genes and these factors were subjected to gene ontology analysis. Next, RT-PCR was performed on a subset of candidate genes to validate the ChRO-seq data. We then performed Masson’s trichrome, H&E, and IHC staining on these tissues to investigate the morphological features of HSA tumor tissue as well as the expression patterns of several proteins identified in our ChRO-seq analysis. Results ChRO-seq analysis revealed over a thousand differentially expressed genes in HSA tissue compared with normal splenic tissue (FDR <0.005). Interestingly the majority of genes overexpressed in HSA tumor tissue were associated with extracellular matrix (ECM) remodeling. This observation correlated well with our histological analysis, which found that HSA tumors contain a rich and complex collagen network. Additionally, we characterized the protein expression patterns of two highly overexpressed molecules identified in ChRO-seq analysis, podoplanin (PDPN) and laminin alpha 4 (LAMA4). We found that the expression of these two ECM-associated factors appeared to be largely limited to transformed endothelial cells within the HSA lesions. Conclusion Outcomes from this study suggest that ECM remodeling has an important role in HSA progression. Additionally, our study identified two potential novel biomarkers of HSA, PDPN and LAMA4. Interestingly, given that function-blocking anti-PDPN have shown anti-tumor effects in mouse models of canine melanoma, our studies raise the possibility that these types of therapeutic strategies could potentially be developed for treating canine HSA.

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