Cannabinoid during adolescence phenocopies reelin haploinsufficiency in prefrontal cortex synapses

In humans and rodents, the protracted development of the prefrontal cortex (PFC) throughout adolescence represents a time for marked vulnerability towards environmental adversities, such as stress or drug exposure. We previously showed that the extracellular matrix protein reelin is an instrumental synaptic modulator that shapes medial PFC (mPFC) circuitry during maturation and is a critical mediator of the vulnerability to environmental stress. Emerging evidence highlight the role of the endocannabinoid system in the postnatal maturation of the PFC and reelin deficiency influences behavioral abnormalities caused by heavy consumption of THC during adolescence. Could the reelin-dependent maturation of prefrontal networks may be vulnerable to cannabinoid exposure during adolescence? To explore this hypothesis, we studied the effects of a single in-vivo exposure to a synthetic cannabinoid on reelin expression and mPFC functions in adolescent male mice. The results show that a single cannabinoid exposure mimics reelin haploinsufficiency by decreasing prefrontal reelin expression in a layer-specific pattern without changing its transcriptional levels. Furthermore, this treatment impeded synaptic plasticity: adolescent cannabinoid lowered long-term potentiation to the magnitude observed in age-matched reelin haploinsufficient males. Quantitative PCR analysis showed that changes in the mRNA levels of NMDARs does not account for the reduction of TBS-LTP. Together, the data show that exposure to cannabinoid during adolescence phenocopies reelin haploinsufficiency and further identifies reelin as a key component of the vulnerability of PFC to environmental insults.

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Glucocorticoids upregulate tropoelastin expression during late stages of fetal lung development

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1995.268.3.l491 ◽

1995 ◽

Vol 268 (3) ◽

pp. L491-L500 ◽

Cited By ~ 17

Author(s):

R. A. Pierce ◽

W. I. Mariencheck ◽

S. Sandefur ◽

E. C. Crouch ◽

W. C. Parks

Keyword(s):

Lung Development ◽

Matrix Protein ◽

Fetal Lung ◽

Extracellular Matrix Protein ◽

Transferase Activity ◽

Mrna Levels ◽

Pregnant Rats ◽

Fetal Lung Development ◽

Chloramphenicol Acetyl Transferase Activity

The production of elastin, an essential extracellular matrix protein of terminal airway interstitium, occurs mostly during early development. Because glucocorticoids influence airway maturation, we studied the effect of dexamethasone (Dex) on tropoelastin expression during fetal lung development. Timed-pregnant rats were treated with Dex (1 mg/kg daily), and fetal lungs were collected 3 days later at 17, 19, and 21 days of gestation. Dex treatment resulted in about a threefold increase in tropoelastin mRNA levels at 19 days concomitant with accelerated airway development. By in situ hybridization, Dex treatment increased the number of tropoelastin-expressing cells and the level of tropoelastin mRNA per cell. In organ culture, Dex increased lung tropoelastin expression and augmented cortisol stimulation of tropoelastin expression. In fetal pulmonary artery smooth muscle cells, 10(-8) M Dex upregulated tropoelastin mRNA expression and increased tropoelastin promoter-chloramphenicol acetyl transferase activity in transient transfections. These data indicate that pharmacologically administered glucocorticoids transcriptionally upregulate fetal lung tropoelastin expression and suggest that steroid hormones may be important regulators of elastin production in vivo.

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Specific heparan sulfate modifications stabilize the synaptic organizer MADD-4/Punctin at C. elegans neuromuscular junctions

Genetics ◽

10.1093/genetics/iyab073 ◽

2021 ◽

Author(s):

Mélissa Cizeron ◽

Laure Granger ◽

Hannes E BÜlow ◽

Jean-Louis Bessereau

Keyword(s):

Heparan Sulfate ◽

Matrix Protein ◽

Neuromuscular Junctions ◽

Core Protein ◽

Extracellular Matrix Protein ◽

Inhibitory Synapses ◽

Single Chain ◽

C Elegans ◽

Sugar Chains

Abstract Heparan sulfate proteoglycans contribute to the structural organization of various neurochemical synapses. Depending on the system, their role involves either the core protein or the glycosaminoglycan chains. These linear sugar chains are extensively modified by heparan sulfate modification enzymes, resulting in highly diverse molecules. Specific modifications of glycosaminoglycan chains may thus contribute to a sugar code involved in synapse specificity. Caenorhabditis elegans is particularly useful to address this question because of the low level of genomic redundancy of these enzymes, as opposed to mammals. Here, we systematically mutated the genes encoding heparan sulfate modification enzymes in C. elegans and analyzed their impact on excitatory and inhibitory neuromuscular junctions. Using single chain antibodies that recognize different heparan sulfate modification patterns, we show in vivo that these two heparan sulfate epitopes are carried by the SDN-1 core protein, the unique C. elegans syndecan orthologue, at neuromuscular junctions. Intriguingly, these antibodies differentially bind to excitatory and inhibitory synapses, implying unique heparan sulfate modification patterns at different neuromuscular junctions. Moreover, while most enzymes are individually dispensable for proper organization of neuromuscular junctions, we show that 3-O-sulfation of SDN-1 is required to maintain wild-type levels of the extracellular matrix protein MADD-4/Punctin, a central synaptic organizer that defines the identity of excitatory and inhibitory synaptic domains at the plasma membrane of muscle cells.

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Low-Intensity Pulsed Ultrasound Promotes Autophagy-Mediated Migration of Mesenchymal Stem Cells and Cartilage Repair

Cell Transplantation ◽

10.1177/0963689720986142 ◽

2021 ◽

Vol 30 ◽

pp. 096368972098614

Author(s):

Peng Xia ◽

Xinwei Wang ◽

Qi Wang ◽

Xiaoju Wang ◽

Qiang Lin ◽

...

Keyword(s):

Cartilage Repair ◽

Matrix Protein ◽

Extracellular Matrix Protein ◽

Histopathological Analysis ◽

Pulsed Ultrasound ◽

Low Intensity Pulsed Ultrasound ◽

Low Intensity ◽

Autophagy Inhibitor

Mesenchymal stem cell (MSC) migration is promoted by low-intensity pulsed ultrasound (LIPUS), but its mechanism is unclear. Since autophagy is known to regulate cell migration, our study aimed to investigate if LIPUS promotes the migration of MSCs via autophagy regulation. We also aimed to investigate the effects of intra-articular injection of MSCs following LIPUS stimulation on osteoarthritis (OA) cartilage. For the in vitro study, rat bone marrow-derived MSCs were treated with an autophagy inhibitor or agonist, and then they were stimulated by LIPUS. Migration of MSCs was detected by transwell migration assays, and stromal cell-derived factor-1 (SDF-1) and C-X-C chemokine receptor type 4 (CXCR4) protein levels were quantified. For the in vivo study, a rat knee OA model was generated and treated with LIPUS after an intra-articular injection of MSCs with autophagy inhibitor added. The cartilage repair was assessed by histopathological analysis and extracellular matrix protein expression. The in vitro results suggest that LIPUS increased the expression of SDF-1 and CXCR4, and it promoted MSC migration. These effects were inhibited and enhanced by autophagy inhibitor and agonist, respectively. The in vivo results demonstrate that LIPUS significantly enhanced the cartilage repair effects of MSCs on OA, but these effects were blocked by autophagy inhibitor. Our results suggest that the migration of MSCs was enhanced by LIPUS through the activation autophagy, and LIPUS improved the protective effect of MSCs on OA cartilage via autophagy regulation.

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Contrasting migratory response of astrocytoma cells to tenascin mediated by different integrins

Journal of Cell Science ◽

10.1242/jcs.109.8.2161 ◽

1996 ◽

Vol 109 (8) ◽

pp. 2161-2168 ◽

Cited By ~ 5

Author(s):

A. Giese ◽

M.A. Loo ◽

S.A. Norman ◽

S. Treasurywala ◽

M.E. Berens

Keyword(s):

Cell Adhesion ◽

Matrix Protein ◽

Glioma Cells ◽

Extracellular Matrix Protein ◽

Integrin Receptors ◽

Migration Assay ◽

Dose Dependent Fashion ◽

Beta 1 Integrin

Tenascin, an extracellular matrix protein, is expressed in human gliomas in vitro and in vivo. The distribution of tenascin at the invasive edge of these tumors, even surrounding solitary invading cells, suggests a role for this protein as a regulator of glioma cell migration. We tested whether purified tenascin, passively deposited on surfaces, influenced the adhesion or migration of a human gliomaderived cell line, SF-767. Adhesion of glioma cells to tenascin increased in a dose-dependent fashion up to a coating concentration of 10 micrograms/ml. Higher coating concentrations resulted in progressively fewer cells attaching. Cell adhesion could be blocked to basal levels using anti-beta 1 integrin antibodies. In contrast, when anti-alpha v antibodies were added to the medium of cells on tenascin, cell adhesion was enhanced slightly. Using a microliter scale migration assay, we found that cell motility on tenascin was dose dependently stimulated at coating concentrations of 1 and 3 micrograms/ml, but migration was inhibited below levels of non-specific motility when tested at coating concentrations of 30 and 100 micrograms/ml. Migration on permissive concentrations of tenascin could be reversibly inhibited with anti-beta 1, while treatment with anti-alpha v antibodies increased migration rates. We conclude that SF-767 glioma cells express two separate integrin receptors that mediate contrasting adhesive and migratory responses to tenascin.

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Initial characterization of anosmin-1, a putative extracellular matrix protein synthesized by definite neuronal cell populations in the central nervous system

Journal of Cell Science ◽

10.1242/jcs.109.7.1749 ◽

1996 ◽

Vol 109 (7) ◽

pp. 1749-1757 ◽

Cited By ~ 5

Author(s):

N. Soussi-Yanicostas ◽

J.P. Hardelin ◽

M.M. Arroyo-Jimenez ◽

O. Ardouin ◽

R. Legouis ◽

...

Keyword(s):

Central Nervous System ◽

Extracellular Matrix ◽

Nervous System ◽

Cell Membrane ◽

Heparan Sulfate ◽

Matrix Protein ◽

Neuronal Cell ◽

Extracellular Matrix Protein ◽

Cell Populations

The KAL gene is responsible for the X-chromosome linked form of Kallmann's syndrome in humans. Upon transfection of CHO cells with a human KAL cDNA, the corresponding encoded protein, KALc, was produced. This protein is N-glycosylated, secreted in the cell culture medium, and is localized at the cell surface. Several lines of evidence indicate that heparan-sulfate chains of proteoglycan(s) are involved in the binding of KALc to the cell membrane. Polyclonal and monoclonal antibodies to the purified KALc were generated. They allowed us to detect and characterize the protein encoded by the KAL gene in the chicken central nervous system at late stages of embryonic development. This protein is synthesized by definite neuronal cell populations including Purkinje cells in the cerebellum, mitral cells in the olfactory bulbs and several subpopulations in the optic tectum and the striatum. The protein, with an approximate molecular mass of 100 kDa, was named anosmin-1 in reference to the deficiency of the sense of smell which characterizes the human disease. Anosmin-1 is likely to be an extracellular matrix component. Since heparin treatment of cell membrane fractions from cerebellum and tectum resulted in the release of the protein, we suggest that one or several heparan-sulfate proteoglycans are involved in the binding of anosmin-1 to the membranes in vivo.

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Stromal protein βig-h3 reprogrammes tumour microenvironment in pancreatic cancer

Gut ◽

10.1136/gutjnl-2018-317570 ◽

2018 ◽

Vol 68 (4) ◽

pp. 693-707 ◽

Cited By ~ 18

Author(s):

Delphine Goehrig ◽

Jérémy Nigri ◽

Rémi Samain ◽

Zhichong Wu ◽

Paola Cappello ◽

...

Keyword(s):

Pancreatic Cancer ◽

Tumour Growth ◽

Matrix Protein ◽

Immune Escape ◽

Interaction Mechanism ◽

Tumour Microenvironment ◽

Extracellular Matrix Protein ◽

Ductal Adenocarcinoma ◽

The Impact

ObjectivePancreatic cancer is associated with an abundant stromal reaction leading to immune escape and tumour growth. This massive stroma drives the immune escape in the tumour. We aimed to study the impact of βig-h3 stromal protein in the modulation of the antitumoural immune response in pancreatic cancer.DesignWe performed studies with p48-Cre;KrasG12D, pdx1-Cre;KrasG12D;Ink4a/Arffl/fl, pdx1-Cre;KrasG12D; p53R172H mice and tumour tissues from patients with pancreatic ductal adenocarcinoma (PDA). Some transgenic mice were given injections of anti-βig-h3, anti-CD8, anti-PD1 depleting antibodies. Tumour growth as well as modifications in the activation of local immune cells were analysed by flow cytometry, immunohistochemistry and immunofluorescence. Tissue stiffness was measured by atomic force microscopy.ResultsWe identified βig-h3 stromal-derived protein as a key actor of the immune paracrine interaction mechanism that drives pancreatic cancer. We found that βig-h3 is highly produced by cancer-associated fibroblasts in the stroma of human and mouse. This protein acts directly on tumour-specific CD8+ T cells and F4/80 macrophages. Depleting βig-h3 in vivo reduced tumour growth by enhancing the number of activated CD8+ T cell within the tumour and subsequent apoptotic tumour cells. Furthermore, we found that targeting βig-h3 in established lesions released the tissue tension and functionally reprogrammed F4/80 macrophages in the tumour microenvironment.ConclusionsOur data indicate that targeting stromal extracellular matrix protein βig-h3 improves the antitumoural response and consequently reduces tumour weight. Our findings present βig-h3 as a novel immunological target in pancreatic cancer.

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Direct observation of structure and dynamics during phase separation of an elastomeric protein

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1701877114 ◽

2017 ◽

Vol 114 (22) ◽

pp. E4408-E4415 ◽

Cited By ~ 78

Author(s):

Sean E. Reichheld ◽

Lisa D. Muiznieks ◽

Fred W. Keeley ◽

Simon Sharpe

Keyword(s):

Phase Separation ◽

Matrix Protein ◽

Molecular Mechanisms ◽

Bulk Water ◽

Extracellular Matrix Protein ◽

Elastic Fibers ◽

Cross Linking ◽

Structure And Dynamics ◽

Chemical Cross Linking

Despite its growing importance in biology and in biomaterials development, liquid–liquid phase separation of proteins remains poorly understood. In particular, the molecular mechanisms underlying simple coacervation of proteins, such as the extracellular matrix protein elastin, have not been reported. Coacervation of the elastin monomer, tropoelastin, in response to heat and salt is a critical step in the assembly of elastic fibers in vivo, preceding chemical cross-linking. Elastin-like polypeptides (ELPs) derived from the tropoelastin sequence have been shown to undergo a similar phase separation, allowing formation of biomaterials that closely mimic the material properties of native elastin. We have used NMR spectroscopy to obtain site-specific structure and dynamics of a self-assembling elastin-like polypeptide along its entire self-assembly pathway, from monomer through coacervation and into a cross-linked elastic material. Our data reveal that elastin-like hydrophobic domains are composed of transient β-turns in a highly dynamic and disordered chain, and that this disorder is retained both after phase separation and in elastic materials. Cross-linking domains are also highly disordered in monomeric and coacervated ELP3 and form stable helices only after chemical cross-linking. Detailed structural analysis combined with dynamic measurements from NMR relaxation and diffusion data provides direct evidence for an entropy-driven mechanism of simple coacervation of a protein in which transient and nonspecific intermolecular hydrophobic contacts are formed by disordered chains, whereas bulk water and salt are excluded.

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Integrin-αvβ3 regulates thrombopoietin-mediated maintenance of hematopoietic stem cells

Blood ◽

10.1182/blood-2011-02-335430 ◽

2012 ◽

Vol 119 (1) ◽

pp. 83-94 ◽

Cited By ~ 45

Author(s):

Terumasa Umemoto ◽

Masayuki Yamato ◽

Jun Ishihara ◽

Yoshiko Shiratsuchi ◽

Mika Utsumi ◽

...

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

Matrix Protein ◽

Ex Vivo ◽

Extracellular Matrix Protein ◽

Αvβ3 Integrin ◽

Hematopoietic Stem ◽

Β3 Integrin ◽

Ex Vivo Culture

AbstractThroughout life, one's blood supply depends on sustained division of hematopoietic stem cells (HSCs) for self-renewal and differentiation. Within the bone marrow microenvironment, an adhesion-dependent or -independent niche system regulates HSC function. Here we show that a novel adhesion-dependent mechanism via integrin-β3 signaling contributes to HSC maintenance. Specific ligation of β3-integrin on HSCs using an antibody or extracellular matrix protein prevented loss of long-term repopulating (LTR) activity during ex vivo culture. The actions required activation of αvβ3-integrin “inside-out” signaling, which is dependent on thrombopoietin (TPO), an essential cytokine for activation of dormant HSCs. Subsequent “outside-in” signaling via phosphorylation of Tyr747 in the β3-subunit cytoplasmic domain was indispensable for TPO-dependent, but not stem cell factor-dependent, LTR activity in HSCs in vivo. This was accompanied with enhanced expression of Vps72, Mll1, and Runx1, 3 factors known to be critical for maintaining HSC activity. Thus, our findings demonstrate a mechanistic link between β3-integrin and TPO in HSCs, which may contribute to maintenance of LTR activity in vivo as well as during ex vivo culture.

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Effect of cyclosporin A treatment on the in vivo regulation of type I MHC gene expression

Journal of Applied Physiology ◽

10.1152/japplphysiol.00763.2003 ◽

2004 ◽

Vol 97 (2) ◽

pp. 475-483 ◽

Cited By ~ 18

Author(s):

Julia M. Giger ◽

Fadia Haddad ◽

Anqi X. Qin ◽

Kenneth M. Baldwin

Keyword(s):

Cyclosporin A ◽

Promoter Activity ◽

Pcr Analysis ◽

Type I ◽

Mrna Levels ◽

Activated T Cells ◽

Obvious Effect ◽

Enhancer Region ◽

Muscle Gene

Rat soleus muscle consists predominantly of slow type I fibers. We have shown previously through deletion analysis that the highest level of reporter activity that we measure when injecting type I myosin heavy chain (MHC) promoter (MHC1)-linked luciferase plasmid into soleus muscles depends on the presence of a 550-bp upstream enhancer (3,450–2,900) region of the promoter. Because the calcineurin-nuclear factor of activated T cells (NFAT) pathway has been implicated in the regulation of the slow muscle gene program, particularly the MHC1 isoform, and the MHC1 promoter contains several putative NFAT sites, we examined via deletion and mutation analyses whether this pathway is involved in the regulation of promoter activity in soleus. Nine days of treatment with the calcineurin inhibitor cyclosporin A (CsA) caused a significant decrease in activity of the −3,500- and −3,450-bp promoters compared with vehicle-treated rats. Truncation of the promoter to −2,900 bp or smaller reduced the activity and also eliminated the CsA responsiveness, thus implying that the enhancer region is required for CsA responsiveness. Surprisingly, mutating the two NFAT elements within the enhancer region had no obvious effect on promoter activity. CsA treatment resulted in an increase in the mRNA levels of fast-type IIa and IIx MHC isoforms, but RT-PCR analysis of MHC1 pre-mRNA and mature mRNA expression in soleus muscles revealed no differences between vehicle- and CsA-treated rats. Although CsA affects the activity of the MHC1 promoter, it appears that its effect is not through direct binding of NFAT to sites on the promoter.

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Differential response to myocardial reperfusion injury in eNOS-deficient mice

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00855.2001 ◽

2002 ◽

Vol 282 (6) ◽

pp. H2422-H2426 ◽

Cited By ~ 55

Author(s):

Brent R. Sharp ◽

Steven P. Jones ◽

David M. Rimmer ◽

David J. Lefer

Keyword(s):

Ischemia Reperfusion ◽

Myocardial Necrosis ◽

In Vivo Model ◽

Pcr Analysis ◽

Inos Mrna ◽

Mrna Levels ◽

Wild Type ◽

Significant Difference ◽

Deficient Mice

Two strains of endothelial nitric oxide synthase (eNOS)-deficient (−/−) mice have been developed that respond differently to myocardial ischemia-reperfusion (MI/R). We evaluated both strains of eNOS−/− mice in an in vivo model of MI/R. Harvard (Har) eNOS−/− mice ( n = 12) experienced an 84% increase in myocardial necrosis compared with wild-type controls ( P < 0.05). University of North Carolina (UNC) eNOS−/−( n = 10) exhibited a 52% reduction in myocardial injury versus wild-type controls ( P < 0.05). PCR analysis of myocardial inducible NO synthase (iNOS) mRNA levels revealed a significant ( P < 0.05) increase in the UNC eNOS−/− mice compared with wild-type mice, and there was no significant difference between the Har eNOS−/− and wild-type mice. UNC eNOS−/− mice treated with an iNOS inhibitor (1400W) exacerbated the extent of myocardial necrosis. When treated with 1400W, Har eNOS−/− did not exhibit a significant increase in myocardial necrosis. These data demonstrate that two distinct strains of eNOS−/− mice display opposite responses to MI/R. Although the protection seen in the UNC eNOS−/− mouse may result from compensatory increases in iNOS, other genes may be involved.

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