Overexpressed Gliotactin activates BMP signaling through interfering with the Tkv–Dad association

Genome ◽  
2020 ◽  
pp. 1-12
Author(s):  
Zohreh Sharifkhodaei ◽  
Vanessa J. Auld
Keyword(s):  
Mrna Level ◽  
Ectopic Expression ◽  
Inhibitory Effect ◽  
Bmp Signaling ◽  
Type I ◽  
Protein Levels ◽  
Barrier Formation ◽  
Epithelial Junctions ◽  
Pupal Wing ◽  
And Function

Epithelial junctions ensure cell–cell adhesion and establish permeability barriers between cells. At the corners of epithelia, the tricellular junction (TCJ) is formed by three adjacent epithelial cells and generates a functional barrier. In Drosophila, a key TCJ protein is Gliotactin (Gli) where loss of Gli disrupts barrier formation and function. Conversely, overexpressed Gli spreads away from the TCJ and triggers apoptosis, delamination, and cell migration. Thus, Gli protein levels are tightly regulated and by two mechanisms, at the protein levels by tyrosine phosphorylation and endocytosis and at the mRNA level through microRNA-184. Regulation of Gli mRNA is mediated through a Gli–BMP–miR184 feedback loop. Excessive Gli triggers BMP signaling pathway through the activation of Tkv type-I BMP receptor and Mad. Elevated level of pMad induces micrRNA-184 expression which in turn targets the Gli 3′UTR and mRNA degradation. Gli activation of Tkv is not through its ligand Dpp but rather through the inhibition of Dad, an inhibitory-Smad. Here, we show that ectopic expression of Gli interferes with Tkv–Dad association by sequestering Dad away from Tkv. The reduced inhibitory effect of Dad on Tkv results in the increased Tkv–pMad signaling activity, and this effect is continuous through larval and pupal wing formation.

Endogenous patterns of TGFbeta superfamily signaling during early Xenopus development

Development ◽  
2000 ◽  
Vol 127 (13) ◽  
pp. 2917-2931 ◽  
Author(s):  
S. Faure ◽  
M.A. Lee ◽  
T. Keller ◽  
P. ten Dijke ◽  
M. Whitman
Keyword(s):  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Ectopic Expression ◽  
Xenopus Embryo ◽  
Bmp Signaling ◽  
Type I ◽  
Embryonic Cells ◽  
Xenopus Development ◽  
Zygotic Transcription ◽  
Carboxy Terminal

Transforming growth factor beta (TGFbeta) superfamily signaling has been implicated in patterning of the early Xenopus embryo. Upon ligand stimulation, TGFbeta receptors phosphorylate Smad proteins at carboxy-terminal SS(V/M)S consensus motifs. Smads 1/5/8, activated by bone morphogenetic protein (BMP) signaling, induce ventral mesoderm whereas Smad2, activated by activin-like ligands, induces dorsal mesoderm. Although ectopic expression studies are consistent with roles for TGFbeta signals in early Xenopus embryogenesis, when and where BMP and activin-like signaling pathways are active endogenously has not been directly examined. In this study, we investigate the temporal and spatial activation of TGFbeta superfamily signaling in early Xenopus development by using antibodies specific for the type I receptor-phosphorylated forms of Smad1/5/8 and Smad2. We find that Smad1/5/8 and two distinct isoforms of Smad2, full-length Smad2 and Smad2(delta)exon3, are phosphorylated in early embryos. Both Smad1/5/8 and Smad2/Smad2(delta)exon3 are activated after, but not before, the mid-blastula transition (MBT). Endogenous activation of Smad2/Smad2(delta)exon3 requires zygotic transcription, while Smad1/5/8 activation at MBT appears to involve transcription-independent regulation. We also find that the competence of embryonic cells to respond to TGF(delta) superfamily ligands is temporally regulated and may be a determinant of early patterning. Levels of phospho-Smad1/5/8 and of phospho-Smad2/Smad2(delta)exon3 are asymmetrically distributed across both the animal-vegetal and dorsoventral axes. The timing of the development of these asymmetries differs for phospho-Smad1/5/8 and for phospho-Smad2/Smad2(delta)exon3, and the spatial distribution of phosphorylation of each Smad changes dramatically as gastrulation begins. We discuss the implications of our results for endogenous functions of BMP and activin-like signals as candidate morphogens regulating primary germ layer formation and dorsoventral patterning of the early Xenopus embryo.

scholarly journals Discoidin Domain Receptor 1 Protein Is a Novel Modulator of Megakaryocyte-Collagen Interactions

2013 ◽  
Vol 288 (23) ◽  
pp. 16738-16746 ◽  
Author(s):  
Vittorio Abbonante ◽  
Cristian Gruppi ◽  
Diana Rubel ◽  
Oliver Gross ◽  
Remigio Moratti ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Type I ◽  
Discoidin Domain Receptor ◽  
Glycoprotein Vi ◽  
Protein Levels ◽  
Collagen Receptors ◽  
And Function ◽  
Extracellular Matrix Receptors ◽  
Collagen Receptor ◽  
Discoidin Domain Receptor 1

Growing evidence demonstrates that extracellular matrices regulate many aspects of megakaryocyte (MK) development; however, among the different extracellular matrix receptors, integrin α2β1 and glycoprotein VI are the only collagen receptors studied in platelets and MKs. In this study, we demonstrate the expression of the novel collagen receptor discoidin domain receptor 1 (DDR1) by human MKs at both mRNA and protein levels and provide evidence of DDR1 involvement in the regulation of MK motility on type I collagen through a mechanism based on the activity of SHP1 phosphatase and spleen tyrosine kinase (Syk). Specifically, we demonstrated that inhibition of DDR1 binding to type I collagen, preserving the engagement of the other collagen receptors, glycoprotein VI, α2β1, and LAIR-1, determines a decrease in MK migration due to the reduction in SHP1 phosphatase activity and consequent increase in the phosphorylation level of its main substrate Syk. Consistently, inhibition of Syk activity restored MK migration on type I collagen. In conclusion, we report the expression and function of a novel collagen receptor on human MKs, and we point out that an increasing level of complexity is necessary to better understand MK-collagen interactions in the bone marrow environment.

scholarly journals Sex differences and similarities in the neuroimmune response to central administration of poly I:C

2020 ◽  
Author(s):  
Caitlin K. Posillico ◽  
Rosa E. Garcia-Hernandez ◽  
Natalie C. Tronson
Keyword(s):  
Sex Differences ◽  
Neurological Diseases ◽  
Type I Interferons ◽  
Poly I:C ◽  
Type I ◽  
Central Administration ◽  
Protein Levels ◽  
Males And Females ◽  
And Function ◽  
Neuroimmune Activation

ABSTRACTThe neuroimmune system is required for normal neural processes, including modulation of cognition, emotion, and adaptive behaviors. Aberrant neuroimmune activation is associated with dysregulation of memory and emotion, though the precise mechanisms at play are complex and highly context dependent. Sex differences neuroimmune activation and function further complicate our understanding of its roles in cognitive and affective regulation. Here, we characterized the physiological sickness and inflammatory response of the hippocampus following intracerebroventricular (ICV) administration of a synthetic viral mimic, polyinosinic:polycytidylic acid (poly I:C), in both male and female C57Bl/6 mice. We observed that poly I:C induced weight loss, fever, and elevations of cytokine and chemokines in the hippocampus of both sexes. Specifically, we found transient increases in gene expression and protein levels of IL-1a, IL-1β, IL-4, IL-6, TNFa, CCL2, and CXCL10, where males showed a greater magnitude of response compared with females. Only males showed increased IFNa and IFNγ in response to poly I:C, whereas both males and females exhibited elevations of IFNβ, demonstrating a specific sex difference in the anti-viral response in the hippocampus. This suggests that type I interferons are one potential node mediating sex-specific cytokine responses and neuroimmune effects on synaptic plasticity and cognition. These findings highlight the importance of using both males and females and analyzing a broad set of inflammatory markers in order to identify the precise, sex-specific roles for neuroimmune dysregulation in neurological diseases and disorders including Alzheimer’s Disease and depression.

Abstract 503: Regulation of D5 Dopamine Receptor on Renalase Expression and Function in Rat Renal Proximal Tubule Cells

Hypertension ◽  
2012 ◽  
Vol 60 (suppl_1) ◽  
Author(s):  
Shaoxing Wang ◽  
Pedro Jose ◽  
Chunyu Zeng
Keyword(s):  
Proximal Tubule ◽  
Dopamine Receptor ◽  
Sch 23390 ◽  
Mrna Level ◽  
Inhibitory Effect ◽  
Renal Proximal Tubule ◽  
Wky Rats ◽  
Wistar Kyoto ◽  
And Function

The dopaminergic and sympathetic systems interact to regulate blood pressure. Our previous studies show the regulation of dopamine receptor on α 1 -adrenergic receptor function. Due to the regulation of renalase on sympathetic tone, we hypothesize that dopamine receptor, especially D 1 -like receptor, might regulate renalase in kidney. The effect of D 1 -like receptor on renalase expression and function was checked in immortalized renal proximal tubule (RPT) cells from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs). It resulted that D 1 -like receptor agonist, fenoldopam (10 -7 -10 -5 M), increased renalase protein expression and function in WKY RPT cells, in contrast, decreased it in SHR cells. These effects were blocked by D 1 -like receptor antagonist SCH 23390 (10 -6 M). Fenoldopam increased renalase mRNA level in WKY RPT cells, but not in SHR cells. Fenoldopam increased the degradation of renalase protein in both WKY and SHR cells. However, the degradation degree was higher in SHR cells than in WKY cells. The regulation of D 1 -like receptor on renalase was mainly via D 5 receptor, because inhibition of D 5 , not D 1 receptor, by antisense blocked inhibitory effect of D 1 -like receptor on renalase in WKY cells. Moreover, inhibition of PKC, by PKC inhibitor 19-31, blocked the effect of fenoldopam on renalase expression; stimulation of PKC, by PKC agonist (PMA), inhibited renalase expression and function, indicating that PKC is involved in the process. Consistent with the in-vitro study, renalase expression was lower in kidney from SHRs than in WKY rats. It indicated that D 1 -like receptor, via D 5 receptor, regulates renalase expression and function in RPT cells, aberrant regulation of D 5 receptor on renalase might be involved in the pathogenesis of hypertension.

scholarly journals Deletion of Alox15 improves kidney dysfunction and inhibits fibrosis by increased PGD2 in the kidney

2021 ◽  
Vol 25 (5) ◽  
pp. 445-455
Author(s):  
Naohiro Takahashi ◽  
Hiroaki Kikuchi ◽  
Ayaka Usui ◽  
Taisuke Furusho ◽  
Takuya Fujimaru ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Interstitial Fibrosis ◽  
Mrna Level ◽  
Type I ◽  
Renal Tubules ◽  
Mrna Levels ◽  
Metabolizing Enzymes ◽  
Protein Levels ◽  
Lipid Metabolizing Enzymes

Abstract Background Lipid-metabolizing enzymes and their metabolites affect inflammation and fibrosis, but their roles in chronic kidney disease (CKD) have not been completely understood. Methods To clarify their role in CKD, we measured the mRNA levels of major lipid-metabolizing enzymes in 5/6 nephrectomized (Nx) kidneys of C57BL/6 J mice. Mediator lipidomics was performed to reveal lipid profiles of CKD kidneys. Results In 5/6 Nx kidneys, both mRNA and protein levels of Alox15 were higher when compared with those in sham kidneys. With respect to in situ hybridization, the mRNA level of Alox15 was higher in renal tubules of 5/6 Nx kidneys. To examine the role of Alox15 in CKD pathogenesis, we performed 5/6 Nx on Alox15−/− mice. Alox15−/− CKD mice exhibited better renal functions than wild-type mice. Interstitial fibrosis was also inhibited in Alox15−/− CKD mice. Mediator lipidomics revealed that Alox15−/− CKD mouse kidneys had significantly higher levels of PGD2 than the control. To investigate the effects of PGD2 on renal fibrosis, we administered PGD2 to TGF-β1-stimulated NRK-52E cells and HK-2 cells, which lead to a dose-dependent suppression of type I collagen and αSMA in both cell lines. Conclusion Increased PGD2 in Alox15−/− CKD mouse kidneys could inhibit fibrosis, thereby resulting in CKD improvement. Thus, Alox15 inhibition and PGD2 administration may be novel therapeutic targets for CKD.

scholarly journals T-614 Promotes Osteoblastic Cell Differentiation by Increasing Dlx5 Expression and Regulating the Activation of p38 and NF-κB

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Jinglue Song ◽  
Hongli Liu ◽  
Qi Zhu ◽  
Yutong Miao ◽  
Feiyan Wang ◽  
...  
Keyword(s):  
Ectopic Expression ◽  
Bone Destruction ◽  
Collagen Type I ◽  
Inhibitory Effect ◽  
Osteoblastic Differentiation ◽  
Nodule Formation ◽  
Osteoblastic Cell ◽  
Type I ◽  
Autoimmune Inflammatory Disease

Rheumatoid arthritis (RA) is an autoimmune inflammatory disease characterized by bone loss. Degree of inflammation has been identified as an important initiator of skeletal damage in RA. Iguratimod (T-614) is an anti-inflammatory agent which has been reported to show the inhibitory effect of bone destruction in RA. However, the role of T-614 in osteoblast differentiation is still not clear. In this study, we intended to find the effect of T-614 on the osteogenesis process. We detected osteogenesis markers and transcription factors associated with osteoblastic lineage and bone formation in the culture of mesenchymal stem cells which differentiate osteoblast. The contents and activity of alkaline phosphatase, levels of collagen type I and bone gla protein, and calcium nodule formation were increased significantly after T-614 treated. Meanwhile, the mRNAs expressions of Osterix and Dlx5 were also found to be increased significantly by real-time PCR. The changes of levels of phosphorylation of p38 and NF-κB were also detected by Western blot. The results showed that T-614 promotes osteoblastic differentiation by increasing the expression of Osterix and Dlx5 and increasing the activation of P38. T-614 could advance the ectopic expression of NF-κB to suppress inflammation, which indirectly inhibits the damage of the osteoblasts.

Differential regulation of G protein expression in rat hearts exposed to chronic hypoxia

1995 ◽  
Vol 269 (6) ◽  
pp. H1865-H1873 ◽  
Author(s):  
R. Kacimi ◽  
J. M. Moalic ◽  
A. Aldashev ◽  
D. E. Vatner ◽  
J. P. Richalet ◽  
...  
Keyword(s):  
Gene Expression ◽  
G Protein ◽  
Functional Activity ◽  
Protein Level ◽  
Chronic Hypoxia ◽  
Mrna Level ◽  
Mrna Levels ◽  
Protein Levels ◽  
Rat Hearts ◽  
And Function

Chronic hypoxia impairs adrenergic responsiveness. A modulation of Gs and/or G1 protein alpha-subunits may be associated with the downregulation of the beta-adrenergic receptors previously found in chronic hypoxia. G protein gene expression and protein level and function in rat hearts exposed to a 30-day hypobaric chronic hypoxia were compared with control rat hearts. No change was observed in G alpha s mRNA levels in either right or left ventricles. In right ventricles, mRNA levels of G alpha i-2 increased by 40% (P < 0.05), but not in left ventricles. In both left and right ventricles, chronic hypoxia did not modify G alpha i-2 and G alpha s protein amounts, but significantly decreased functional activity of G alpha s. In conclusion, gene expression, protein levels of G alpha s and G alpha i-2, and activity of G alpha s do not change in parallel fashion with chronic hypoxia. In chronic hypoxic right ventricles, although the mRNA level of G alpha i-2 is increased, the protein level is unchanged. One potential mechanism of desensitization to catecholamines in chronic hypoxia appears to involve a decreased functional activity of G alpha s in spite of normal mRNA and protein levels

scholarly journals The Regulation and Function of the Forkhead Transcription Factor, Forkhead Box O1, Is Dependent on the Progesterone Receptor in Endometrial Carcinoma

Endocrinology ◽  
2007 ◽  
Vol 149 (4) ◽  
pp. 1942-1950 ◽  
Author(s):  
Erin C. Ward ◽  
Anna V. Hoekstra ◽  
Leen J. Blok ◽  
P. Hanifi-Moghaddam ◽  
John R. Lurain ◽  
...  
Keyword(s):  
Endometrial Cancer ◽  
Progesterone Receptor ◽  
Cell Lines ◽  
Type I ◽  
Ishikawa Cell ◽  
Protein Levels ◽  
Forkhead Box ◽  
Ishikawa Cells ◽  
Presence And Absence ◽  
And Function

In many type I endometrial cancers, the PTEN gene is inactivated, which ultimately leads to constitutively active Akt and the inhibition of Forkhead box O1 (FOXO1), a member of the FOXO subfamily of Forkhead/winged helix family of transcription factors. The expression, regulation, and function of FOXO1 in endometrial cancer were investigated in this study. Immunohistochemical analysis of 49 endometrial tumor tissues revealed a decrease of FOXO1 expression in 95.9% of the cases compared with the expression in normal endometrium. In four different endometrial cancer cell lines (ECC1, Hec1B, Ishikawa, and RL95), FOXO1 mRNA was expressed at similar levels; however, protein levels were low or undetectable in Ecc1, Ishikawa, and RL95 cells. Using small interfering RNA technology, we demonstrated that the low levels of FOXO1 protein were due to the involvement of Skp2, an oncogenic subunit of the Skp1/Cul1/F-box protein ubiquitin complex, given that silencing Skp2 increased FOXO1 protein expression in Ishikawa cells. Inhibition of Akt in Ishikawa cells also increased nuclear FOXO1 protein levels. Additionally, progestins increased FOXO1 protein levels, specifically through progesterone receptor B (PRB) as determined by using stably transfected PRA-specific and PRB-specific Ishikawa cell lines. Finally, overexpression of triple mutant (Tm) FOXO1 in the PR-specific Ishikawa cell lines caused cell cycle arrest and significantly decreased proliferation in the presence and absence of the progestin, R5020. Furthermore, TmFOXO1 overexpression induced apoptosis in PRB-specific cells in the presence and absence of ligand. Taken together, these data provide insight into the phosphoinositide-3-kinase/Akt/FOXO pathway for the determination of progestin responsiveness and the development of alternate therapies for endometrial cancer.

MicroRNA miR-24 inhibits erythropoiesis by targeting activin type I receptor ALK4

Blood ◽  
2008 ◽  
Vol 111 (2) ◽  
pp. 588-595 ◽  
Author(s):  
Qiang Wang ◽  
Zheng Huang ◽  
Huiling Xue ◽  
Chengcheng Jin ◽  
Xiu-Li Ju ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Ectopic Expression ◽  
K562 Cells ◽  
Erythroid Differentiation ◽  
Hematopoietic Progenitor Cells ◽  
Type I ◽  
Hematopoietic Progenitor ◽  
Protein Levels ◽  
Smad2 Phosphorylation ◽  
Differentiation Marker

MicroRNAs have been suggested to modulate a variety of cellular events. Here we report that miR-24 regulates erythroid differentiation by influencing the expression of human activin type I receptor ALK4 (hALK4). Ectopic expression of miR-24 reduces the mRNA and protein levels of hALK4 by targeting the 3′-untranslated region of hALK4 mRNA and interferes with activin-induced Smad2 phosphorylation and reporter expression. Furthermore, miR-24 represses the activin-mediated accumulation of hemoglobin, an erythroid differentiation marker, in erythroleukemic K562 cells and decreases erythroid colony-forming and burst-forming units of CD34+ hematopoietic progenitor cells. ALK4 expression is inversely correlated with miR-24 expression during the early stages of erythroid differentiation, and the forced expression of miR-24 leads to a delay of activin-induced maturation of hematopoietic progenitor cells in liquid culture. Thus, our findings define a regulation mode of miR-24 on erythropoiesis by impeding ALK4 expression.

scholarly journals MicroRNA bta-miR-365-3p inhibits proliferation but promotes differentiation of primary bovine myoblasts by targeting the activin A receptor type I

2020 ◽  
Author(s):  
Dan Hao ◽  
Xiaogang Wang ◽  
Xiao Wang ◽  
Bo Thomsen ◽  
Xianyong Lan ◽  
...  
Keyword(s):  
Muscle Development ◽  
Target Genes ◽  
Heart Tissue ◽  
Nuclear Antigen ◽  
Type I ◽  
Differentiation Markers ◽  
Protein Coding ◽  
Protein Levels ◽  
Mrna Targets ◽  
And Function

Abstract Background MicroRNAs act as post-transcriptional regulators that repress translation or degrades mRNA transcripts. Each microRNA has many mRNA targets and each mRNA may be targeted by several microRNAs. Skeletal muscles express a plethora of microRNA genes that regulate muscle development and function by controlling the expression of protein-coding target genes. To expand our understanding of the role of microRNA, specifically bta-miR-365-3p, in muscle biology, and we studied its function to primary bovine myoblast proliferation and differentiation. Results We first show that bta-miR-365-3p is predominantly expressed in skeletal muscle and heart tissue in Chinese Qinchuan beef cattle. Quantitative PCR and western blotting showed that overexpression of bta-miR-365-3p significantly reduced the levels of cyclinD1 (CCND1), cyclin dependent kinase 2 (CDK2) and proliferating cell nuclear antigen (PCNA) but stimulated the expression of muscle differentiation markers MYOD1, MYOG. Moreover, downregulation of bta-miR-365-3p increased expression of CCND1, CDK2 and PCNA but decreased expression of MYOD1 and MYOG at both mRNA and protein levels. Furthermore, flow cytometry, EdU proliferation assays and immunostaining showed that increased levels of bta-miR-365-3p suppressed cell proliferation but promoted myotube formation, whereas a decreased level of bta-miR-365-3p had opposite consequences. Finally, we determined that ACVR1 is a direct target of bta-miR-365-3p. Thus, dual luciferase gene reporter assays demonstrated that bta-miR-365-3p can bind to the 3'UTR of ACVR1 to regulate its expression. Consistently, knock-down of ACVR1 was associated with reduced CDK2, CCND1 and PCNA expression but increased MYOD1 expression. Conclusion Collectively these data suggest that bta-miR-365-3p represses proliferation but promotes differentiation of bovine myoblasts through a mechanism involving downregulation of ACVR1.

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