Cav-1 Protein Levels in Serum and Infarcted Brain Correlate with Hemorrhagic Volume in a Mouse Model of Thromboembolic Stroke, Independently of rt-PA Administration

Background The mechanisms underlying fibromyalgia (FM) pain are not understood. The US Food and Drug Administration has recommended three drugs for treating FM—namely, pregabalin, duloxetine and milnacipran; however, these medications are associated with severe side effects. Objective To create a mouse model of FM pain using dual injections of acidic saline to cause mechanical hyperalgesia and test whether ASIC3, Nav1.7 and Nav1.8 are involved in this process and whether electroacupuncture (EA) can reverse these phenomena. Methods The FM model was established by injecting acidic saline twice into 40 ICR mice. The mice were assigned to subgroups (n=8 each) treated with different EA frequencies (2, 15 and 50 Hz). ASIC3, Nav1.7 and Nav1.8 expression levels were measured by Western blotting and immunohistochemistry. Results Significant mechanical hyperalgesia was induced on day 8 in FM mice, which was reversed by 2, 15 and 50 Hz EA. ASIC3, Nav1.7 and Nav1.8 protein levels increased significantly in both the dorsal root ganglion and in the spinal cord of FM model mice. These changes were further attenuated by 2, 15 and 50 Hz EA. Conclusion Reduced nociceptive ASIC3, Nav1.7 and Nav1.8 proteins are involved in the preventive effects of EA against FM, and this series of molecules may represent targets for FM treatment.

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Abstract 285: Osteopontin Deficiency Ameliorates Heart Failure with Preserved Ejection Fraction Pathology by Upregulating Mitochondrial 2-Oxoglutarate Dehydrogenase Like Enzyme

Circulation Research ◽

10.1161/res.121.suppl_1.285 ◽

2017 ◽

Vol 121 (suppl_1) ◽

Author(s):

Keyvan Yousefi ◽

Wen Ding ◽

Lina A Shehadeh

Keyword(s):

Mouse Model ◽

Diastolic Dysfunction ◽

Interstitial Fibrosis ◽

Matricellular Protein ◽

Double Knockout ◽

Protein Levels ◽

Lower Protein ◽

Myocardial Energetics ◽

First Time

HFpEF is an increasingly prevalent syndrome associated with impaired myocardial energetics, for which no etiologic therapy is available. Osteopontin (OPN) is a matricellular protein that is upregulated in the circulation of HFpEF patients, and reported to induce mitochondrial stress in rodent cardiomyocytes. Here we evaluate the role of circulating OPN in regulating myocardial function in the nephrotic Col4a3 -/- mouse model of HFpEF. We performed extensive cardiac, biochemical and mitochondrial analyses of the Col4a3 -/- mouse and found a striking HFpEF phenotype. We showed OPN levels were elevated in Col4a3 -/- mice (FC=2.1, n=6; p<.01). Col4a3 -/- mice were hypertensive, had diastolic dysfunction, myocyte hypertrophy and interstitial fibrosis - all of which were ameliorated in Col4a3 -/- OPN -/- mice (n=5-20; p<.05). Col4a3 -/- hearts had dysmorphic mitochondria (EM), lowered antioxidant capacity as a 50% reduction in GSH/GSSG ratio (n=6; p<.05) and lower protein levels of mitochondrial respiratory complexes I, II and IV (p<.05). Flux assay in adult cardiomyocytes showed that maximal respiration was reduced in Col4a3 -/- hearts (575.84±37.6 vs 322.34±25.48 pmol/min in WT, n=9; p<.0001). Microarray data (validated by mitochondrial blot) implicated OGDHL as decreased in Col4a3 -/- hearts but increased in double knockout Col4a3 -/- OPN -/- hearts compared to WT (n=3; p<.05). OGDH activity was also lower in Col4a3 -/- hearts (17.1±7.3 vs 2.5±1.1 mU/mg in WT; n=6; p<.05). In Col4a3 -/- mice, heart-specific AAV9-mediated overexpression of OGDHL, similar to global OPN KO, improved survival by ~50-100% (p<.0001). Isovolumetric relaxation time, a marker of diastolic dysfunction, which is prolonged in Col4a3 -/- mice (26.17 vs 15.30±1 ms, n=26; p<.001) was decreased in Col4a3 -/- OPN -/- mice (18.1±1 ms, n=37; p<.01) as well as in AAV9-cTnT-OGDHL-treated Col4a3 -/- mice (16.7±2.5 ms, n=8; p<.05). In conclusion, we present a new mouse model for HFpEF in which diastolic function and lifespan can be improved by genetic deletion of OPN or cardiac OGDHL gene therapy. Our results elucidate for the first time the pivotal roles of circulating OPN and cardiac OGDHL in HFpEF pathophysiology and present two related potential therapeutic targets for HFpEF.

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Expression of XBP1s in B lymphocytes is critical for pristane-induced lupus nephritis in mice

AJP Renal Physiology ◽

10.1152/ajprenal.00472.2019 ◽

2020 ◽

Vol 318 (5) ◽

pp. F1258-F1270 ◽

Cited By ~ 1

Author(s):

Li Xiang ◽

An Liu ◽

Guoshuang Xu

Keyword(s):

B Cells ◽

Lupus Nephritis ◽

Mouse Model ◽

B Cell ◽

Lupus Erythematosus ◽

Cell Activation ◽

B Lymphocyte ◽

B Cell Activation ◽

Protein Levels ◽

Almost All

B lymphocyte hyperactivity plays a pathogenic role in systemic lupus erythematosus (SLE), and spliced X box-binding protein 1 (XBP1s) has been implicated in B cell maturation and differentiation. We hypothesized that blockade of the XBP1s pathway inhibits the B cell hyperactivity underlying SLE and lupus nephritis (LN) development. In the present study, we systematically evaluated the changes in B cell activation induced by the Xbp1 splicing inhibitor STF083010 in a pristane-induced lupus mouse model. The lupus mouse model was successfully established, as indicated by the presence of LN with markedly increased urine protein levels, renal deposition of Ig, and mesangial cell proliferation. In lupus mice, B cell hyperactivity was confirmed by increased CD40 and B cell-activating factor levels. B cell activation and plasma cell overproduction were determined by increases in CD40-positive and CD138-positive cells in the spleens of lupus mice by flow cytometry and further confirmed by CD45R and Ig light chain staining in the splenic tissues of lupus mice. mRNA and protein expression of XBP1s in B cells was assessed by real-time PCR, Western blot analysis, and immunofluorescence analysis and was increased in lupus mice. In addition, almost all changes were reversed by STF083010 treatment. However, the expression of XBP1s in the kidneys did not change when mice were exposed to pristane and STF083010. Taken together, these findings suggest that expression of XBP1s in B cells plays key roles in SLE and LN development. Blockade of the XBP1s pathway may be a potential strategy for SLE and LN treatment.

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MicroRNA-186 improves fracture healing through activating the bone morphogenetic protein signalling pathway by inhibiting SMAD6 in a mouse model of femoral fracture

Bone and Joint Research ◽

10.1302/2046-3758.811.bjr-2018-0251.r1 ◽

2019 ◽

Vol 8 (11) ◽

pp. 550-562 ◽

Cited By ~ 3

Author(s):

C. Wang ◽

G-F. Zheng ◽

X-F. Xu

Keyword(s):

Mouse Model ◽

Fracture Healing ◽

Bone Morphogenetic Protein ◽

Femoral Fracture ◽

Maximum Load ◽

Bone Volume ◽

Signalling Pathway ◽

Protein Levels ◽

Morphogenetic Protein ◽

Bmp Signalling

Objectives MicroRNAs (miRNAs) have been reported as key regulators of bone formation, signalling, and repair. Fracture healing is a proliferative physiological process where the body facilitates the repair of a bone fracture. The aim of our study was to explore the effects of microRNA-186 (miR-186) on fracture healing through the bone morphogenetic protein (BMP) signalling pathway by binding to Smad family member 6 (SMAD6) in a mouse model of femoral fracture. Methods Microarray analysis was adopted to identify the regulatory miR of SMAD6. 3D micro-CT was performed to assess the bone volume (BV), bone volume fraction (BVF, BV/TV), and bone mineral density (BMD), followed by a biomechanical test for maximum load, maximum radial degrees, elastic radial degrees, and rigidity of the femur. The positive expression of SMAD6 in fracture tissues was measured. Moreover, the miR-186 level, messenger RNA (mRNA) level, and protein levels of SMAD6, BMP-2, and BMP-7 were examined. Results MicroRNA-186 was predicted to regulate SMAD6. Furthermore, SMAD6 was verified as a target gene of miR-186. Overexpressed miR-186 and SMAD6 silencing resulted in increased callus formation, BMD and BV/TV, as well as maximum load, maximum radial degrees, elastic radial degrees, and rigidity of the femur. In addition, the mRNA and protein levels of SMAD6 were decreased, while BMP-2 and BMP-7 levels were elevated in response to upregulated miR-186 and SMAD6 silencing. Conclusion In conclusion, the study indicated that miR-186 could activate the BMP signalling pathway to promote fracture healing by inhibiting SMAD6 in a mouse model of femoral fracture. Cite this article: Bone Joint Res 2019;8:550–562.

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Modification of an in vivo SCID mouse model of human angiogenesis that allows for the differential genomic molecular analysis of RNA and protein levels in endothelial cells undergoing angiogenesis

International Journal of Radiation Oncology*Biology*Physics ◽

10.1016/s0360-3016(00)80340-5 ◽

2000 ◽

Vol 48 (3) ◽

pp. 270

Author(s):

M.K Khan ◽

S.A Linn ◽

M Sutorik ◽

C.L Addison ◽

T.S Lawrence ◽

...

Keyword(s):

Endothelial Cells ◽

Mouse Model ◽

Molecular Analysis ◽

Scid Mouse ◽

Protein Levels ◽

Scid Mouse Model

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Glial inhibitors influence the mRNA and protein levels of mGlu2/3, 5 and 7 receptors and potentiate the analgesic effects of their ligands in a mouse model of neuropathic pain

Pain ◽

10.1016/j.pain.2009.09.002 ◽

2009 ◽

Vol 147 (1) ◽

pp. 175-186 ◽

Cited By ~ 29

Author(s):

Maria Osikowicz ◽

Malgorzata Skup ◽

Joanna Mika ◽

Wioletta Makuch ◽

Julita Czarkowska-Bauch ◽

...

Keyword(s):

Neuropathic Pain ◽

Mouse Model ◽

Protein Levels ◽

Analgesic Effects

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Reduced activity-dependent protein levels in a mouse model of the fragile X premutation

Neurobiology of Learning and Memory ◽

10.1016/j.nlm.2014.01.011 ◽

2014 ◽

Vol 109 ◽

pp. 160-168 ◽

Cited By ~ 5

Author(s):

Ramona E. von Leden ◽

Lindsey C. Curley ◽

Gian D. Greenberg ◽

Michael R. Hunsaker ◽

Rob Willemsen ◽

...

Keyword(s):

Mouse Model ◽

Fragile X ◽

Protein Levels ◽

Fragile X Premutation ◽

Dependent Protein ◽

Activity Dependent ◽

Reduced Activity

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Small molecule inhibitors and a kinase-dead expressing mouse model demonstrate that the kinase activity of Chk1 is essential for mouse embryos and cancer cells

10.1101/2020.01.21.913467 ◽

2020 ◽

Author(s):

Somsundar V Muralidharan ◽

Lisa M Nilsson ◽

Mattias F Lindberg ◽

Jonas A Nilsson

Keyword(s):

T Cells ◽

Mouse Model ◽

Cancer Cells ◽

Null Allele ◽

Protein Levels ◽

Conditional Allele ◽

Car T ◽

High Level ◽

Atr Kinase ◽

Chk1 Kinase

AbstractChk1 kinase is downstream of the ATR kinase in the sensing of improper replication. Previous cell culture studies have demonstrated that Chk1 is essential for replication and Chk1 inhibitors are efficacious against tumors with high-level replication stress such as Myc-induced lymphoma cells. Treatment with Chk1 inhibitors also combines well with certain chemotherapeutic drugs and effects associates with induction of DNA damage and reduction of Chk1 protein levels. Most studies of Chk1 function has relied on the use of inhibitors. Whether or not a mouse or cancer cells could survive if a kinase-dead form of Chk1 is expressed has not been investigated before. Here we generate a mouse model that expresses a kinase-dead (D130A) allele in the mouse germline. We find that this mouse is overtly normal and does not have problems with erythropoiesis with ageing as previously been shown for a mouse expressing one null allele. However, similar to a null allele, homozygous kinase-dead mice cannot be generated and timed pregnancies of heterozygous mice suggest lethality of homozygous blastocysts at around the time of implantation. By breeding the kinase-dead Chk1 mouse with a conditional allele we are able to demonstrate that expression of only one kinase-dead allele, but no wildtype allele, of Chek1 is lethal for Myc-induced cancer cells. Finally, treatment of melanoma cells with tumor-infiltrating T cells or CAR-T cells is effective even if Chk1 is inhibited, suggesting that Chk1 inhibitors can be safely administered in patients where immunotherapy is an essential component of the arsenal against cancer.

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