36 results on '"Jianlin Zhang"'
Search Results
2. Cardiomyocyte Expression of ZO-1 Is Essential for Normal Atrioventricular Conduction but Does Not Alter Ventricular Function
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Andrew D. McCulloch, Angela K. Peter, Debbie Ferng, Kirk U. Knowlton, Yunghang Chan, Selina Manying Huang, Matthew Klos, Hongqiang Cheng, Yusu Gu, Robert S. Ross, Ju Chen, Ruixia Li, Jordan Kelly Towne, Kirk L. Peterson, Jianlin Zhang, Nancy D. Dalton, and Kevin P. Vincent
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Male ,Scaffold protein ,cardiac ,Physiology ,1.1 Normal biological development and functioning ,Clinical Sciences ,coxsackie adenovirus receptor ,Cardiorespiratory Medicine and Haematology ,Cardiovascular ,Connexins ,Article ,NAV1.5 Voltage-Gated Sodium Channel ,Mice ,conduction system ,Underpinning research ,Transcription (biology) ,atrioventricular block ,medicine ,Animals ,Ventricular Function ,2.1 Biological and endogenous factors ,Aetiology ,zonula occludens ,Myocytes ,biology ,Ventricular function ,vinculin ,Chemistry ,Atrioventricular conduction ,Vinculin ,Cadherins ,medicine.disease ,Cell biology ,Heart Disease ,Cardiovascular System & Hematology ,Atrioventricular Node ,Zonula Occludens-1 Protein ,biology.protein ,Electrical conduction system of the heart ,Signal transduction ,Cardiology and Cardiovascular Medicine ,Atrioventricular block ,alpha Catenin - Abstract
Rationale: ZO-1 (Zonula occludens-1), a plasma membrane-associated scaffolding protein regulates signal transduction, transcription, and cellular communication. Global deletion of ZO-1 in the mouse is lethal by embryonic day 11.5. The function of ZO-1 in cardiac myocytes (CM) is largely unknown. Objective: To determine the function of CM ZO-1 in the intact heart, given its binding to other CM proteins that have been shown instrumental in normal cardiac conduction and function. Methods and Results: We generated ZO-1 CM-specific knockout (KO) mice using α-Myosin Heavy Chain-nuclear Cre (ZO-1cKO) and investigated physiological and electrophysiological function by echocardiography, surface ECG and conscious telemetry, intracardiac electrograms and pacing, and optical mapping studies. ZO-1cKO mice were viable, had normal Mendelian ratios, and had a normal lifespan. Ventricular morphometry and function were not significantly different between the ZO-1cKO versus control (CTL) mice, basally in young or aged mice, or even when hearts were subjected to hemodynamic loading. Atrial mass was increased in ZO-1cKO. Electrophysiological and optical mapping studies indicated high-grade atrioventricular (A-V) block in ZO-1cKO comparing to CTL hearts. While ZO-1-associated proteins such as vinculin, connexin 43, N-cadherin, and α-catenin showed no significant change with the loss of ZO-1, Connexin-45 and Coxsackie-adenovirus (CAR) proteins were reduced in atria of ZO-1cKO. Further, with loss of ZO-1, ZO-2 protein was increased significantly in ventricular CM in a presumed compensatory manner but was still not detected in the AV nodal myocytes. Importantly, the expression of the sodium channel protein NaV1.5 was altered in AV nodal cells of the ZO-1cKO versus CTL. Conclusions: ZO-1 protein has a unique physiological role in cardiac nodal tissue. This is in alignment with its known interaction with CAR and Cx45, and a new function in regulating the expression of NaV1.5 in AV node. Uniquely, ZO-1 is dispensable for function of the working myocardium.
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- 2020
3. Fate alteration of bone marrow-derived macrophages ameliorates kidney fibrosis in murine model of unilateral ureteral obstruction
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Ziyan Zhang, Ying Yang, Hai-Long Wang, Xiaojian Feng, Xi Qiao, Xinyan Liu, Ying Wang, Jianlin Zhang, Lihua Wang, Guoping Zheng, Linxia Zhao, Min Hu, Rui Guo, and Qi Cao
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Male ,medicine.medical_treatment ,Anti-Inflammatory Agents ,030232 urology & nephrology ,Inflammation ,030204 cardiovascular system & hematology ,Kidney ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Transforming Growth Factor beta ,Fibrosis ,medicine ,Renal fibrosis ,Animals ,beta Catenin ,Transplantation ,business.industry ,Macrophages ,medicine.disease ,Mice, Inbred C57BL ,Disease Models, Animal ,medicine.anatomical_structure ,Cytokine ,Nephrology ,Cancer research ,Cytokines ,Bone marrow ,medicine.symptom ,business ,Myofibroblast ,Signal Transduction ,Ureteral Obstruction ,Transforming growth factor - Abstract
BackgroundRenal fibrosis is a key pathological feature and final common pathway leading to end-stage kidney failure in many chronic kidney diseases. Myofibroblast is the master player in renal fibrosis. However, myofibroblasts are heterogeneous. Recent studies show that bone marrow-derived macrophages transform into myofibroblasts by transforming growth factor (TGF)-β-induced macrophage–myofibroblast transition (MMT) in renal fibrosis.MethodsTGF-β signaling was redirected by inhibition of β-catenin/T-cell factor (TCF) to increase β-catenin/Foxo in bone marrow-derived macrophages. A kidney fibrosis model of unilateral ureteral obstruction was performed in EGFP bone marrow chimera mouse. MMT was examined by flow cytometry analysis of GFP+F4/80+α-SMA+ cells from unilateral ureteral obstruction (UUO) kidney, and by immunofluorescent staining of bone marrow-derived macrophages in vitro. Inflammatory and anti-inflammatory cytokines were analysis by enzyme-linked immunosorbent assay.ResultsInhibition of β-catenin/TCF by ICG-001 combined with TGF-β1 treatment increased β-catenin/Foxo1, reduced the MMT and inflammatory cytokine production by bone marrow-derived macrophages, and thereby, reduced kidney fibrosis in the UUO model.ConclusionsOur results demonstrate that diversion of β-catenin from TCF to Foxo1-mediated transcription not only inhibits the β-catenin/TCF-mediated fibrotic effect of TGF-β, but also enhances its anti-inflammatory action, allowing therapeutic use of TGF-β to reduce both inflammation and fibrosis at least partially by changing the fate of bone marrow-derived macrophages.
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- 2018
4. O-linked β-N-acetylglucosamine transferase plays an essential role in heart development through regulating angiopoietin-1
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Ju Chen, Yongxin Mu, Houzhi Yu, Tongbin Wu, Jianlin Zhang, Sylvia M. Evans, and Firulli, Anthony B
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Cancer Research ,Embryology ,Organogenesis ,Immunostaining ,QH426-470 ,Inbred C57BL ,Cardiovascular ,Mice ,0302 clinical medicine ,Animal Cells ,Medicine and Health Sciences ,Morphogenesis ,Transferase ,Myocytes, Cardiac ,Genetics (clinical) ,Coronary Arteries ,Cells, Cultured ,Cardiomyocytes ,Staining ,0303 health sciences ,Cultured ,Heart development ,Heart ,Animal Models ,Arteries ,Congenital Heart Defects ,Cell biology ,Heart Disease ,Experimental Organism Systems ,Coronary vessel ,Knockout mouse ,cardiovascular system ,Heart Development ,Anatomy ,Cellular Types ,Cardiac ,Research Article ,Cells ,1.1 Normal biological development and functioning ,Muscle Tissue ,Cardiology ,Mouse Models ,Biology ,Research and Analysis Methods ,N-Acetylglucosaminyltransferases ,O-Linked β-N-acetylglucosamine ,Atrial septal defects ,03 medical and health sciences ,Model Organisms ,Underpinning research ,Congenital Disorders ,Angiopoietin-1 ,Genetics ,Animals ,Birth Defects ,cardiovascular diseases ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics ,Heart Disease - Coronary Heart Disease ,030304 developmental biology ,Fetus ,Myocytes ,Muscle Cells ,Embryos ,RNA ,Biology and Life Sciences ,Cell Biology ,Mice, Inbred C57BL ,Biological Tissue ,Specimen Preparation and Treatment ,Cardiovascular Anatomy ,Animal Studies ,Ventricular Septal Defects ,Blood Vessels ,Organism Development ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
O-linked N-acetylglucosamine (GlcNAc) transferase (OGT) is the only enzyme catalyzing O-GlcNAcylation. Although it has been shown that OGT plays an essential role in maintaining postnatal heart function, its role in heart development remains unknown. Here we showed that loss of OGT in early fetal cardiomyocytes led to multiple heart developmental defects including hypertrabeculation, biventricular dilation, atrial septal defects, ventricular septal defects, and defects in coronary vessel development. In addition, RNA sequencing revealed that Angiopoietin-1, required within cardiomyocytes for both myocardial and coronary vessel development, was dramatically downregulated in cardiomyocyte-specific OGT knockout mouse hearts. In conclusion, our data demonstrated that OGT plays an essential role in regulating heart development through activating expression of cardiomyocyte Angiopoietin-1., Author summary As the first functional organ to form, the heart develops from a simple tube to a complex organ with four chambers to keep up with the body’s increasing demand for blood. During this period, myocardium, the major component of heart, undergoes substantial dynamic architectural remodeling at different levels. This remodeling also coincides with coronary vessel development to form a functional heart. Defects in architectural remodeling of myocardium or coronary vessel development can lead to anatomical changes during heart development, resulting in congenital heart disease. O-linked N-acetylglucosamine (GlcNAc) transferase (OGT) is the only enzyme catalyzing protein O-GlcNAcylation using UDP-GlcNAc, the end product of the hexosamine biosynthetic pathway. Hence, OGT is also considered a sensor of cellular nutrient levels. In this study, we found that loss of OGT in myocardium disrupted myocardial remodeling, as well as coronary vessel development, through regulating Angiopoietin-1 expression. Our study not only demonstrates an essential role for myocardial OGT during heart development, but also has potential implications for the development of new treatment options targeting coronary diseases.
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- 2020
5. BACH1 is transcriptionally inhibited by TET1 in hepatocellular carcinoma in a microRNA-34a-dependent manner to regulate autophagy and inflammation
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Rongge Cao, Xuehu Sun, Hongmei Zhu, Xingyu Wang, and Jianlin Zhang
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0301 basic medicine ,Carcinoma, Hepatocellular ,Blotting, Western ,Cell ,Apoptosis ,Inflammation ,Biology ,Real-Time Polymerase Chain Reaction ,medicine.disease_cause ,Mixed Function Oxygenases ,Metastasis ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,Cell Line, Tumor ,Proto-Oncogene Proteins ,Autophagy ,medicine ,Animals ,Humans ,Cell Proliferation ,Oligonucleotide Array Sequence Analysis ,Pharmacology ,Mice, Inbred BALB C ,Cell Cycle ,Liver Neoplasms ,medicine.disease ,digestive system diseases ,Gene Expression Regulation, Neoplastic ,MicroRNAs ,Basic-Leucine Zipper Transcription Factors ,030104 developmental biology ,medicine.anatomical_structure ,MicroRNA 34a ,030220 oncology & carcinogenesis ,Cancer research ,Cytokines ,medicine.symptom ,Carcinogenesis ,Neoplasm Transplantation - Abstract
Hepatocellular carcinoma (HCC), one of the main contributors to cancer-associated deaths globally, is characterized by high invasiveness. Herein, we studied the molecular mechanisms underlying ten-eleven translocation 1 (TET1)-mediated autophagy in HCC. Following data mining using GSE101728, GSE14520 and GSE138178, TET1 was screened out, and the differential expression of TET1 was verified by bioinformatics analysis. TET1, one of the prognostic markers in HCC, was poorly expressed in HCC. Through functional experiments, we determined that upregulation of TET1 inhibited the proliferation, migration, invasion, tumorigenesis, metastasis and inflammatory factors of HCC cells, and promoted cell autophagy and apoptosis. Mechanistically, TET1 activated miR-34a by demethylating miR-34a. BTB domain and CNC homology 1 (BACH1) was identified as the target gene of miR-34a. Notably, Downregulation of miR-34a increased cellular inflammatory factors and decreased autophagy in the presence of TET1, while declines in BACH1 suppressed cellular inflammatory factors and enhanced autophagy in the presence of miR-34a inhibitor. BACH1 negatively regulated the p53 pathway. In conclusion, TET1 is a tumor suppressor in the progression of HCC by regulating the miR-34a/BACH1/p53 axis, and may contribute to the improvement of HCC prognosis and therapy.
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- 2021
6. Nesprin 1α2 is essential for mouse postnatal viability and nuclear positioning in skeletal muscle
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Wei Feng, Xi Fang, Jennifer Veevers, Ju Chen, Jianlin Zhang, Larry Gerace, and Matthew J. Stroud
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0301 basic medicine ,LINC complex ,Muscle Fibers, Skeletal ,Kinesins ,Inbred C57BL ,Medical and Health Sciences ,Mice ,Myofibrils ,Nuclear protein ,Cytoskeleton ,Research Articles ,Mice, Knockout ,Nuclear Proteins ,Kinesin ,Skeletal ,Biological Sciences ,Cell biology ,Phenotype ,medicine.anatomical_structure ,psychological phenomena and processes ,Protein Binding ,Signal Transduction ,Genotype ,Knockout ,Nerve Tissue Proteins ,Biology ,Muscle Fibers ,03 medical and health sciences ,Report ,mental disorders ,medicine ,Animals ,Protein Interaction Domains and Motifs ,Actin ,Cell Nucleus ,Binding Sites ,Nesprin ,Skeletal muscle ,Cell Biology ,Actins ,Mice, Inbred C57BL ,Cytoskeletal Proteins ,Cell nucleus ,030104 developmental biology ,Mutation ,Myofibril ,Developmental Biology - Abstract
Defects in nuclear positioning occur in muscle diseases and correlate with muscle dysfunction. In this study, Stroud et al. show that nesprin 1α2 is the fundamental nesprin 1 isoform for nuclear positioning, skeletal muscle function, and postnatal viability., The position of the nucleus in a cell is controlled by interactions between the linker of nucleoskeleton and cytoskeleton (LINC) complex and the cytoskeleton. Defects in nuclear positioning and abnormal aggregation of nuclei occur in many muscle diseases and correlate with muscle dysfunction. Nesprin 1, which includes multiple isoforms, is an integral component of the LINC complex, critical for nuclear positioning and anchorage in skeletal muscle, and is thought to provide an essential link between nuclei and actin. However, previous studies have yet to identify which isoform is responsible. To elucidate this, we generated a series of nesprin 1 mutant mice. We showed that the actin-binding domains of nesprin 1 were dispensable, whereas nesprin 1α2, which lacks actin-binding domains, was crucial for postnatal viability, nuclear positioning, and skeletal muscle function. Furthermore, we revealed that kinesin 1 was displaced in fibers of nesprin 1α2–knockout mice, suggesting that this interaction may play an important role in positioning of myonuclei and functional skeletal muscle.
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- 2017
7. Erythropoietin promotes abdominal aortic aneurysms in mice through angiogenesis and inflammatory infiltration
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Meng Zhang, Wenhai Sui, Cheng Cheng, Fei Xue, Zhenyu Tian, Jing Cheng, Jie Zhang, Tao Zhang, Jianlin Zhang, Weiwei Wang, Wenjing Xiong, Panpan Hao, Jing Ma, Xingli Xu, Shuangxi Wang, Shangwen Sun, Yun Zhang, and Cheng Zhang
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,Angiogenesis ,medicine.drug_class ,education ,macromolecular substances ,030204 cardiovascular system & hematology ,Monoclonal antibody ,03 medical and health sciences ,Aortic aneurysm ,Mice ,0302 clinical medicine ,Apolipoproteins E ,hemic and lymphatic diseases ,medicine.artery ,medicine ,Animals ,Humans ,cardiovascular diseases ,Aorta, Abdominal ,Erythropoietin ,Mice, Knockout ,Aorta ,business.industry ,Vascular disease ,Angiotensin II ,Endothelial Cells ,General Medicine ,medicine.disease ,Abdominal aortic aneurysm ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,cardiovascular system ,business ,medicine.drug ,Aortic Aneurysm, Abdominal - Abstract
Abdominal aortic aneurysm (AAA) is a potentially fatal vascular disease, but the underlying mechanisms remain unknown. Here, we tested the hypothesis that erythropoietin (EPO) may promote the formation of AAA. We found that EPO dose-dependently promoted the formation of AAA in both Apoe−/− (66.7%) and wild-type (WT) (60%) mice receiving a high dose of EPO. EPO monoclonal antibodies given to Apoe−/− mice receiving angiotensin II (AngII) stimulation resulted in a markedly lower incidence of AAA (from 86.7 to 20%, P
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- 2019
8. Kindlin-2 is Essential for Preserving Integrity of the Developing Heart and Preventing Ventricular Rupture
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Paola Cattaneo, Jennifer Veevers, Yangzhao Zhou, Zhiyuan Zhang, Yongxin Mu, Angela K. Peter, Robert S. Ross, Ana Maria Manso, Jianlin Zhang, Xinmin Zhou, Kirk U. Knowlton, Sylvia M. Evans, and Ju Chen
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Pathology ,medicine.medical_specialty ,Heart Ventricles ,Integrin ,Developing heart ,Cardiac metabolism ,Embryonic Development ,Muscle Proteins ,Article ,Extracellular matrix ,Mice ,Physiology (medical) ,medicine ,Myocyte ,Animals ,Myocytes, Cardiac ,Basement membrane ,Mice, Knockout ,Rupture ,biology ,business.industry ,Integrin beta1 ,Myocardium ,Myocardium metabolism ,Embryo ,Embryo, Mammalian ,Extracellular Matrix ,Cytoskeletal Proteins ,medicine.anatomical_structure ,biology.protein ,Cardiology and Cardiovascular Medicine ,business - Published
- 2019
9. RBFox2-miR-34a-Jph2 axis contributes to cardiac decompensation during heart failure
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Ju Chen, Chen Gao, Changwei Shao, Chaoliang Wei, Jianlin Zhang, Yuanchao Xue, Lan-Tao Gou, Shuxun Ren, Yibin Wang, Xiang-Dong Fu, Yu Zhou, Jing Hu, and Yajing Hao
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medicine.medical_specialty ,Down-Regulation ,Muscle Proteins ,RNA-binding protein ,chemistry.chemical_compound ,Mice ,Internal medicine ,JPH2 ,microRNA ,medicine ,Animals ,Humans ,Antagomir ,Myocytes, Cardiac ,Heart Failure ,Mice, Knockout ,Multidisciplinary ,business.industry ,Mechanism (biology) ,Membrane Proteins ,Heart ,medicine.disease ,MicroRNAs ,chemistry ,Cardiac decompensation ,PNAS Plus ,Heart failure ,Cardiology ,RNA Splicing Factors ,Regulatory Pathway ,business - Abstract
Heart performance relies on highly coordinated excitation-contraction (EC) coupling, and defects in this critical process may be exacerbated by additional genetic defects and/or environmental insults to cause eventual heart failure. Here we report a regulatory pathway consisting of the RNA binding protein RBFox2, a stress-induced microRNA miR-34a, and the essential EC coupler JPH2. In this pathway, initial cardiac defects diminish RBFox2 expression, which induces transcriptional repression of miR-34a, and elevated miR-34a targets Jph2 to impair EC coupling, which further manifests heart dysfunction, leading to progressive heart failure. The key contribution of miR-34a to this process is further established by administrating its mimic, which is sufficient to induce cardiac defects, and by using its antagomir to alleviate RBFox2 depletion-induced heart dysfunction. These findings elucidate a potential feed-forward mechanism to account for a critical transition to cardiac decompensation and suggest a potential therapeutic avenue against heart failure.
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- 2019
10. Analyzing Oxygen Consumption Rate in Primary Cultured Mouse Neonatal Cardiomyocytes Using an Extracellular Flux Analyzer
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Anne N. Murphy, Aleksander Andreyev, Yoshitake Cho, Oliver R. Zhang, Ana Maria Manso, Jianlin Zhang, Cameron Hill, Ruixia Li, Robert S. Ross, Sarah V. Schurr, Chao Chen, and Shizuko Tachibana
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0301 basic medicine ,General Chemical Engineering ,Mitochondrion ,Cardiovascular ,Mice ,2.1 Biological and endogenous factors ,Psychology ,Myocytes, Cardiac ,Aetiology ,Cells, Cultured ,screening and diagnosis ,Cultured ,Chemistry ,General Neuroscience ,Cardiac myocyte ,Cardiac muscle ,Cell biology ,Mouse neonatal cardiomyocytes ,mitochondria ,Detection ,medicine.anatomical_structure ,Heart Disease ,5.1 Pharmaceuticals ,Medicine ,Cognitive Sciences ,Development of treatments and therapeutic interventions ,Cardiac ,Intracellular ,Biotechnology ,Cardiac function curve ,Cells ,heart ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Oxygen Consumption ,Extracellular ,medicine ,Animals ,extracellular flux analyzer ,Heart Disease - Coronary Heart Disease ,Myocytes ,030102 biochemistry & molecular biology ,General Immunology and Microbiology ,medicine.disease ,4.1 Discovery and preclinical testing of markers and technologies ,Oxygen ,Issue 144 ,030104 developmental biology ,Heart failure ,Biochemistry and Cell Biology ,Flux (metabolism) ,respiration - Abstract
Mitochondria and oxidative metabolism are critical for maintaining cardiac muscle function. Research has shown that mitochondrial dysfunction is an important contributing factor to impaired cardiac function found in heart failure. By contrast, restoring defective mitochondrial function may have beneficial effects to improve cardiac function in the failing heart. Therefore, studying the regulatory mechanisms and identifying novel regulators for mitochondrial function could provide insight which could be used to develop new therapeutic targets for treating heart disease. Here, cardiac myocyte mitochondrial respiration is analyzed using a unique cell culture system. First, a protocol has been optimized to rapidly isolate and culture high viability neonatal mouse cardiomyocytes. Then, a 96-well format extracellular flux analyzer is used to assess the oxygen consumption rate of these cardiomyocytes. For this protocol, we optimized seeding conditions and demonstrated that neonatal mouse cardiomyocytes oxygen consumption rate can be easily assessed in an extracellular flux analyzer. Finally, we note that our protocol can be applied to a larger culture size and other studies, such as intracellular signaling and contractile function analysis.
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- 2019
11. GRP78-targeted ferritin nanocaged ultra-high dose of doxorubicin for hepatocellular carcinoma therapy
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Xiyun Yan, Bing Jiang, Meng Zhou, Chunyi Hao, Jianlin Zhang, Kelong Fan, Ruofei Zhang, Xiuyun Tian, Yaxin Hou, and Xuehui Chen
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Carcinoma, Hepatocellular ,Medicine (miscellaneous) ,Mice, Nude ,Antineoplastic Agents ,02 engineering and technology ,Metastasis ,03 medical and health sciences ,Mice ,0302 clinical medicine ,In vivo ,Carcinoma ,Medicine ,Animals ,Humans ,Doxorubicin ,Tissue Distribution ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Endoplasmic Reticulum Chaperone BiP ,Heat-Shock Proteins ,Mice, Inbred BALB C ,biology ,business.industry ,Liver Neoplasms ,3T3 Cells ,Hep G2 Cells ,021001 nanoscience & nanotechnology ,medicine.disease ,digestive system diseases ,Ferritin ,Drug Liberation ,Targeted drug delivery ,030220 oncology & carcinogenesis ,Hepatocellular carcinoma ,Apoferritins ,biology.protein ,Cancer research ,Nanoparticles ,Female ,Nanocarriers ,0210 nano-technology ,business ,medicine.drug ,Protein Binding ,Research Paper - Abstract
Hepatocellular carcinoma (HCC) remains one of the leading causes of cancer deaths, primarily due to its high incidence of recurrence and metastasis. Considerable efforts have therefore been undertaken to develop effective therapies; however, effective anti-HCC therapies rely on identification of suitable biomarkers, few of which are currently available for drug targeting. Methods: GRP78 was identified as the membrane receptor of HCC-targeted peptide SP94 by immunoprecipitation and mass spectrum analysis. To develop an effective anti-HCC drug nanocarrier, we first displayed GRP78-targeted peptide SP94 onto the exterior surface of Pyrococcus furiosus ferritin Fn (HccFn) by genetic engineering approach, and then loaded doxorubicin (Dox) into the cavities of HccFn via urea-based disassembly/reassembly method, thereby constructing a drug nanocarrier called HccFn-Dox. Results: We demonstrated that HccFn nanocage encapsulated ultra-high dose of Dox (up to 400 molecules Dox/protein nanocage). In vivo animal experiments showed that Dox encapsulated in HccFn-Dox was selectively delivered into HCC tumor cells, and effectively killed subcutaneous and lung metastatic HCC tumors. In addition, HccFn-Dox significantly reduced drug exposure to healthy organs and improved the maximum tolerated dose by six-fold compared with free Dox. Conclusion: In conclusion, our findings clearly demonstrate that GRP78 is an effective biomarker for HCC therapy, and GRP78-targeted HccFn nanocage is effective in delivering anti-HCC drug without damage to healthy tissue.
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- 2018
12. Redirecting TGF-β Signaling through the β-Catenin/Foxo Complex Prevents Kidney Fibrosis
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Yiping Wang, Hai-Long Wang, Guoping Zheng, Min Pang, Yuan Min Wang, Xinrui Tian, Yun Zhang, Chengshi Wang, Min Hu, Hong Yu, Ye Zhao, David Harris, Xin Maggie Wang, Geoff Yu Zhang, Xi Qiao, Lixin Liu, Vincent W. Lee, Qi Cao, Stephen I. Alexander, Jianlin Zhang, Padmashree Rao, and Fei Guo
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0301 basic medicine ,Male ,T cell ,Inflammation ,FOXO1 ,Pyrimidinones ,Kidney ,T-Lymphocytes, Regulatory ,Cell Line ,Transforming Growth Factor beta1 ,03 medical and health sciences ,Mice ,Fibrosis ,medicine ,Animals ,Protein Interaction Domains and Motifs ,Smad3 Protein ,Promoter Regions, Genetic ,Transcription factor ,beta Catenin ,Chemistry ,Forkhead Box Protein O1 ,FOXP3 ,Forkhead Transcription Factors ,General Medicine ,medicine.disease ,Bridged Bicyclo Compounds, Heterocyclic ,Recombinant Proteins ,030104 developmental biology ,medicine.anatomical_structure ,Basic Research ,Nephrology ,Catenin ,Cancer research ,Cytokines ,medicine.symptom ,TCF Transcription Factors ,Transforming growth factor ,Signal Transduction ,Ureteral Obstruction - Abstract
TGF-β is a key profibrotic factor, but targeting TGF-β to prevent fibrosis also abolishes its protective anti-inflammatory effects. Here, we investigated the hypothesis that we can redirect TGF-β signaling by preventing downstream profibrotic interaction of β-catenin with T cell factor (TCF), thereby enhancing the interaction of β-catenin with Foxo, a transcription factor that controls differentiation of TGF-β induced regulatory T cells (iTregs), and thus, enhance anti-inflammatory effects of TGF-β. In iTregs derived from EL4 T cells treated with recombinant human TGF-β1 (rhTGF-β1) in vitro, inhibition of β-catenin/TCF transcription with ICG-001 increased Foxp3 expression, interaction of β-catenin and Foxo1, binding of Foxo1 to the Foxp3 promoter, and Foxo transcriptional activity. Moreover, the level of β-catenin expression positively correlated with the level of Foxo1 binding to the Foxp3 promoter and Foxo transcriptional activity. T cell fate mapping in Foxp3gfp Ly5.1/5.2 mice revealed that coadministration of rhTGF-β1 and ICG-001 further enhanced the expansion of iTregs and natural Tregs observed with rhTGF-β1 treatment alone. Coadministration of rhTGF-β1 with ICG-001 also increased the number of Tregs and reduced inflammation and fibrosis in the kidney fibrosis models of unilateral ureteric obstruction and ischemia-reperfusion injury. Notably, ICG-001 prevented the fibrosis in distant organs (lung and liver) caused by rhTGF-β1. Together, our results show that diversion of β-catenin from TCF- to Foxo-mediated transcription inhibits the β-catenin/TCF–mediated profibrotic effects of TGF-β while enhancing the β-catenin/Foxo–mediated anti-inflammatory effects. Targeting β-catenin/Foxo may be a novel therapeutic strategy in the treatment of fibrotic diseases that lead to organ failure.
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- 2017
13. Lmo7 is dispensable for skeletal muscle and cardiac function
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Kirk L. Peterson, Mary C. Esparza, Jianlin Zhang, Kirk U. Knowlton, Dieu Hung Lao, Nancy D. Dalton, William H. Bradford, Ju Chen, Richard L. Lieber, Shannon N. Bremner, Indroneal Banerjee, Jennifer Veevers, and Yusu Gu
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musculoskeletal diseases ,Cardiac function curve ,medicine.medical_specialty ,MAP Kinase Signaling System ,Physiology ,Gene Expression ,Biology ,Mice ,Degenerative disease ,Internal medicine ,medicine ,Animals ,X-linked muscular dystrophy ,Muscular dystrophy ,Muscle, Skeletal ,Transcription factor ,Mice, Knockout ,Myocardium ,Cardiac muscle ,Skeletal muscle ,Heart ,Articles ,Cell Biology ,LIM Domain Proteins ,medicine.disease ,Muscular Dystrophy, Emery-Dreifuss ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Endocrinology ,ITGA7 ,Transcription Factors - Abstract
Emery-Dreifuss muscular dystrophy (EDMD) is a degenerative disease primarily affecting skeletal muscles in early childhood as well as cardiac muscle at later stages. EDMD is caused by a number of mutations in genes encoding proteins associated with the nuclear envelope (e.g., Emerin, Lamin A/C, and Nesprin). Recently, a novel protein, Lim-domain only 7 ( lmo7) has been reported to play a role in the molecular pathogenesis of EDMD. Prior in vitro and in vivo studies suggested the intriguing possibility that Lmo7 plays a role in skeletal or cardiac muscle pathophysiology. To further understand the in vivo role of Lmo7 in striated muscles, we generated a novel Lmo7-null ( lmo7−/−) mouse line. Using this mouse line, we examined skeletal and cardiac muscle physiology, as well as the role of Lmo7 in a model of muscular dystrophy and regeneration using the dystrophin-deficient mdx mouse model. Our results demonstrated that lmo7−/− mice had no abnormalities in skeletal muscle morphology, physiological function, or regeneration. Cardiac function was also unaffected. Moreover, we found that ablation of lmo7 in mdx mice had no effect on the observed myopathy and muscular regeneration exhibited by mdx mice. Molecular analyses also showed no changes in dystrophin complex factors, MAPK pathway components, and Emerin levels in lmo7 knockout mice. Taken together, we conclude that Lmo7 is dispensable for skeletal muscle and cardiac physiology and pathophysiology.
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- 2015
14. AMP-Activated Protein Kinase and Sirtuin 1 Coregulation of Cortactin Contributes to Endothelial Function
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Traci L. Marin, Liang Wen, David A. Johnson, Brendan Gongol, Tzu-Pin Shentu, Yinsheng Wang, Fan Zhang, Yi Zhu, Jiao Zhang, John Y.-J. Shyy, Jianlin Zhang, Xiaoli Sun, Ming He, and Gregory G. Geary
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0301 basic medicine ,Male ,Cell ,Pulsatile flow ,AMP-Activated Protein Kinases ,Cardiorespiratory Medicine and Haematology ,Cardiovascular ,Mice ,0302 clinical medicine ,AMP-activated protein kinase ,Sirtuin 1 ,2.1 Biological and endogenous factors ,Phosphorylation ,Aetiology ,Cultured ,biology ,Chemistry ,Nitric Oxide Synthase Type III ,Acetylation ,Cell biology ,Protein Transport ,medicine.anatomical_structure ,Phenotype ,030220 oncology & carcinogenesis ,Pulsatile Flow ,RNA Interference ,Cardiology and Cardiovascular Medicine ,Cortactin ,Signal Transduction ,Biotechnology ,caveolin-1 ,Genotype ,Cells ,Knockout ,Clinical Sciences ,macromolecular substances ,Transfection ,Stress ,03 medical and health sciences ,Apolipoproteins E ,medicine ,Genetics ,Animals ,Humans ,Animal ,Endothelial Cells ,cortactin ,Mechanical ,Atherosclerosis ,Actins ,030104 developmental biology ,Cardiovascular System & Hematology ,Caveolin 1 ,Disease Models ,biology.protein ,Function (biology) - Abstract
Objective— Cortactin translocates to the cell periphery in vascular endothelial cells (ECs) on cortical–actin assembly in response to pulsatile shear stress. Because cortactin has putative sites for AMP-activated protein kinase (AMPK) phosphorylation and sirtuin 1 (SIRT1) deacetylation, we examined the hypothesis that AMPK and SIRT1 coregulate cortactin dynamics in response to shear stress. Approach and Results— Analysis of the ability of AMPK to phosphorylate recombinant cortactin and oligopeptides whose sequences matched AMPK consensus sequences in cortactin pointed to Thr-401 as the site of AMPK phosphorylation. Mass spectrometry confirmed Thr-401 as the site of AMPK phosphorylation. Immunoblot analysis with AMPK siRNA and SIRT1 siRNA in human umbilical vein ECs and EC-specific AMPKα2 knockout mice showed that AMPK phosphorylation of cortactin primes SIRT1 deacetylation in response to shear stress. Immunoblot analyses with cortactin siRNA in human umbilical vein ECs, phospho-deficient T401A and phospho-mimetic T401D mutant, or aceto-deficient (9K/R) and aceto-mimetic (9K/Q) showed that cortactin regulates endothelial nitric oxide synthase activity. Confocal imaging and sucrose-density gradient analyses revealed that the phosphorylated/deacetylated cortactin translocates to the EC periphery facilitating endothelial nitric oxide synthase translocation from lipid to nonlipid raft domains. Knockdown of cortactin in vitro or genetic reduction of cortactin expression in vivo in mice substantially decreased the endothelial nitric oxide synthase–derived NO bioavailability. In vivo, atherosclerotic lesions increase in ApoE – /– /cortactin +/– mice, when compared with ApoE –/– /cortactin +/+ littermates. Conclusions— AMPK phosphorylation of cortactin followed by SIRT1 deacetylation modulates the interaction of cortactin and cortical–actin in response to shear stress. Functionally, this AMPK/SIRT1 coregulated cortactin–F-actin dynamics is required for endothelial nitric oxide synthase subcellular translocation/activation and is atheroprotective.
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- 2016
15. MicroRNA 329 Suppresses Angiogenesis by Targeting CD146
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Dongling Yang, Jianlin Zhang, Yongting Luo, Ping Wang, Di Lu, Lina Song, Shu Xing, Xiyun Yan, Hongxia Duan, and Jing Feng
- Subjects
Vascular Endothelial Growth Factor A ,Angiogenesis ,Neovascularization, Physiologic ,CD146 Antigen ,Mice, SCID ,Retinal Neovascularization ,Biology ,Vascular endothelial growth inhibitor ,Neovascularization ,Mice ,Human Umbilical Vein Endothelial Cells ,medicine ,Animals ,Humans ,Molecular Biology ,Cells, Cultured ,Tube formation ,Base Sequence ,Neovascularization, Pathologic ,Tumor Necrosis Factor-alpha ,Endothelial Cells ,Kinase insert domain receptor ,Articles ,Cell Biology ,Mice, Inbred C57BL ,Endothelial stem cell ,Disease Models, Animal ,MicroRNAs ,Vascular endothelial growth factor A ,HEK293 Cells ,Gene Expression Regulation ,Immunology ,Cancer research ,Female ,medicine.symptom ,Proto-oncogene tyrosine-protein kinase Src - Abstract
CD146, an endothelial biomarker, has been shown to be aberrantly upregulated during pathological angiogenesis and functions as a coreceptor for vascular endothelial growth factor receptor 2 (VEGFR-2) to promote disease progression. However, the regulatory mechanisms of CD146 expression during angiogenesis remain unclear. Using a microRNA screening approach, we identified a novel negative regulator of angiogenesis, microRNA 329 (miR-329), that directly targeted CD146 and inhibited CD146-mediated angiogenesis in vitro and in vivo. Endogenous miR-329 expression was downregulated by VEGF and tumor necrosis factor alpha (TNF-α), resulting in the elevation of CD146 in endothelial cells. Upregulation of CD146 facilitated an endothelial response to VEGF-induced SRC kinase family (SKF)/p38 mitogen-activated protein kinase (MAPK)/NF-κB activation and consequently promoted endothelial cell migration and tube formation. Our animal experiments showed that treatment with miR-329 repressed excessive CD146 expression on blood vessels and significantly attenuated neovascularization in a mouse model of pathological angiogenesis. Our findings provide the first evidence that CD146 expression in angiogenesis is regulated by miR-329 and suggest that miR-329 could present a potential therapeutic tool for the treatment of angiogenic diseases.
- Published
- 2013
16. Matrix metalloproteinase-9 of tubular and macrophage origin contributes to the pathogenesis of renal fibrosis via macrophage recruitment through osteopontin cleavage
- Author
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Stephen I. Alexander, Thian Kui Tan, So Ra Lee, Yuan Min Wang, Ye Zhao, Jianlin Zhang, Vincent W. Lee, Yiping Wang, Changqi Wang, Guoping Zheng, David Harris, Qi Cao, Dong Zheng, Ya Wang, Erik W. Thompson, Tzu-Ting Hsu, and Xinrui Tian
- Subjects
Pathology ,medicine.medical_specialty ,Epithelial-Mesenchymal Transition ,Kidney ,urologic and male genital diseases ,Pathology and Forensic Medicine ,Pathogenesis ,Mice ,Cell Movement ,Fibrosis ,medicine ,Renal fibrosis ,Animals ,Epithelial–mesenchymal transition ,Osteopontin ,Molecular Biology ,Cells, Cultured ,beta Catenin ,Mice, Inbred BALB C ,biology ,urogenital system ,Macrophages ,Kidney metabolism ,Epithelial Cells ,Cell Biology ,medicine.disease ,medicine.anatomical_structure ,Matrix Metalloproteinase 9 ,biology.protein ,Kidney Diseases ,Snail Family Transcription Factors ,Myofibroblast ,Transcription Factors ,Ureteral Obstruction - Abstract
A pro-fibrotic role of matrix metalloproteinase-9 (MMP-9) in tubular cell epithelial-mesenchymal transition (EMT) is well established in renal fibrosis; however studies from our group and others have demonstrated some previously unrecognized complexity of MMP-9 that has been overlooked in renal fibrosis. Therefore, the aim of this study was to determine the expression pattern, origin and the exact mechanism underlying the contribution of MMP-9 to unilateral ureteral obstruction (UUO), a well-established model of renal fibrosis via MMP-9 inhibition. Renal MMP-9 expression in BALB/c mice with UUO was examined on day 1, 3, 5, 7, 9, 11 and 14. To inhibit MMP-9 activity, MMP-2/9 inhibitor or MMP-9-neutralizing antibody was administered daily for 4 consecutive days from day 0-3, 6-9 or 10-13 and tissues harvested at day 14. In UUO, there was a bi-phasic early- and late-stage upregulation of MMP-9 activity. Interestingly, tubular epithelial cells (TECs) were the predominant source of MMP-9 during early stage, whereas TECs, macrophages and myofibroblasts produced MMP-9 during late-stage UUO. Early- and late-stage inhibition of MMP-9 in UUO mice significantly reduced tubular cell EMT and renal fibrosis. Moreover, MMP-9 inhibition caused a significant reduction in MMP-9-cleaved osteopontin and macrophage infiltration in UUO kidney. Our in vitro study showed MMP-9-cleaved osteopontin enhanced macrophage transwell migration and MMP-9 of both primary TEC and macrophage induced tubular cell EMT. In summary, our result suggests that MMP-9 of both TEC and macrophage origin may directly or indirectly contribute to the pathogenesis of renal fibrosis via osteopontin cleavage, which, in turn further recruit macrophage and induce tubular cell EMT. Our study also highlights the time dependency of its expression and the potential of stage-specific inhibition strategy against renal fibrosis.
- Published
- 2013
17. Association of β-catenin with P-Smad3 but not LEF-1 dissociates in vitro profibrotic from anti-inflammatory effects of TGF-β1
- Author
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Changqi Wang, Hong Zhao, Ya Wang, Ye Zhao, Yiping Wang, Bo Niu, J. Guy Lyons, David Harris, Jianlin Zhang, Xinrui Tian, Thian Kui Tan, Qi Cao, Michael Kahn, Vincent W. Lee, Guoping Zheng, So Ra Lee, and Tania Tsatralis
- Subjects
medicine.medical_specialty ,Epithelial-Mesenchymal Transition ,Lymphoid Enhancer-Binding Factor 1 ,Blotting, Western ,Anti-Inflammatory Agents ,SMAD ,Biology ,Cell Line ,Transforming Growth Factor beta1 ,Mice ,Internal medicine ,medicine ,Animals ,Immunoprecipitation ,Smad3 Protein ,beta Catenin ,Gene knockdown ,Reverse Transcriptase Polymerase Chain Reaction ,Wnt signaling pathway ,Cell Biology ,In vitro ,Cell biology ,Blot ,Endocrinology ,Microscopy, Fluorescence ,Cell culture ,Catenin ,Protein Binding ,Transforming growth factor - Abstract
Summary Transforming growth factor β1 (TGF-β1) is known to be both anti-inflammatory and profibrotic. Cross-talk between TGF-β/Smad and Wnt/β-catenin pathways in epithelial–mesenchymal transition (EMT) suggests a specific role for β-catenin in profibrotic effects of TGF-β1. However, no such mechanistic role has been demonstrated for β-catenin in the anti-inflammatory effects of TGF-β1. In the present study, we explored the role of β-catenin in the profibrotic and anti-inflammatory effects of TGF-β1 by using a cytosolic, but not membrane, β-catenin knockdown chimera (F-TrCP-Ecad) and the β-catenin/CBP inhibitor ICG-001. TGF-β1 induced nuclear Smad3/β-catenin complex, but not β-catenin/LEF-1 complex or TOP-flash activity, during EMT of C1.1 (renal tubular epithelial) cells. F-TrCP-Ecad selectively degraded TGF-β1-induced cytoplasmic β-catenin and blocked EMT of C1.1 cells. Both F-TrCP-Ecad and ICG-001 blocked TGF-β1-induced Smad3/β-catenin and Smad reporter activity in C1.1 cells, suggesting that TGF-β1-induced EMT depends on β-catenin binding to Smad3, but not LEF-1 downstream of Smad3, through canonical Wnt. In contrast, in J774 macrophages, the β-catenin level was low and was not changed by interferon-γ (IFN-γ) or lipopolysaccharide (LPS) with or without TGF-β1. TGF-β1 inhibition of LPS-induced TNF-α and IFN-γ-stimulated inducible NO synthase (iNOS) expression was not affected by F-TrCP-Ecad, ICG-001 or by overexpression of wild-type β-catenin in J774 cells. Inhibition of β-catenin by either F-TrCP-Ecad or ICG-001 abolished LiCl-induced TOP-flash, but not TGF-β1-induced Smad reporter, activity in J774 cells. These results demonstrate for the first time that β-catenin is required as a co-factor of Smad in TGF-β1-induced EMT of C1.1 epithelial cells, but not in TGF-β1 inhibition of macrophage activation. Targeting β-catenin may dissociate the TGF-β1 profibrotic and anti-inflammatory effects.
- Published
- 2013
18. Adipocyte-specific loss of PPARγ attenuates cardiac hypertrophy
- Author
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Matthew J. Stroud, Xi Fang, Hsien-Da Huang, Nancy D. Dalton, Ju Chen, Tongbin Wu, Kunfu Ouyang, Yusu Gu, Li Fang, Kirk L. Peterson, Nanping Wang, and Jianlin Zhang
- Subjects
0301 basic medicine ,Male ,Peroxisome proliferator-activated receptor ,Adipose tissue ,Inbred C57BL ,Cardiovascular ,Tuberous Sclerosis Complex 1 Protein ,Muscle hypertrophy ,chemistry.chemical_compound ,Mice ,Adipocyte ,Adipocytes ,2.1 Biological and endogenous factors ,Myocyte ,Myocytes, Cardiac ,Aetiology ,Cells, Cultured ,chemistry.chemical_classification ,Mice, Knockout ,Cultured ,TOR Serine-Threonine Kinases ,Diabetes ,General Medicine ,3T3 Cells ,Heart Disease ,Knockout mouse ,Rosiglitazone ,Cardiac ,medicine.drug ,Research Article ,medicine.medical_specialty ,Cells ,Knockout ,Cardiomegaly ,Biology ,03 medical and health sciences ,Internal medicine ,medicine ,Animals ,PI3K/AKT/mTOR pathway ,Myocytes ,Tumor Suppressor Proteins ,Mice, Inbred C57BL ,PPAR gamma ,MicroRNAs ,030104 developmental biology ,Endocrinology ,chemistry ,Thiazolidinediones - Abstract
Adipose tissue is a key endocrine organ that governs systemic homeostasis. PPARγ is a master regulator of adipose tissue signaling that plays an essential role in insulin sensitivity, making it an important therapeutic target. The selective PPARγ agonist rosiglitazone (RSG) has been used to treat diabetes. However, adverse cardiovascular effects have seriously hindered its clinical application. Experimental models have revealed that PPARγ activation increases cardiac hypertrophy. RSG stimulates cardiac hypertrophy and oxidative stress in cardiomyocyte-specific PPARγ knockout mice, implying that RSG might stimulate cardiac hypertrophy independently of cardiomyocyte PPARγ. However, candidate cell types responsible for RSG-induced cardiomyocyte hypertrophy remain unexplored. Utilizing cocultures of adipocytes and cardiomyocytes, we found that stimulation of PPARγ signaling in adipocytes increased miR-200a expression and secretion. Delivery of miR-200a in adipocyte-derived exosomes to cardiomyocytes resulted in decreased TSC1 and subsequent mTOR activation, leading to cardiomyocyte hypertrophy. Treatment with an antagomir to miR-200a blunted this hypertrophic response in cardiomyocytes. In vivo, specific ablation of PPARγ in adipocytes was sufficient to blunt hypertrophy induced by RSG treatment. By delineating mechanisms by which RSG elicits cardiac hypertrophy, we have identified pathways that mediate the crosstalk between adipocytes and cardiomyocytes to regulate cardiac remodeling.
- Published
- 2016
19. Thymosin Beta 4 Is Dispensable for Murine Cardiac Development and Function
- Author
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Thomas Moore-Morris, Kirk L. Peterson, Indroneal Banerjee, Tao Shen, Jianlin Zhang, Nancy D. Dalton, Sylvia M. Evans, Ju Chen, Yusu Gu, and Stephan Lange
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Male ,Cardiac function curve ,Physiology ,Embryonic Development ,Neovascularization, Physiologic ,Biology ,Article ,Small hairpin RNA ,Mice ,medicine ,Animals ,Gene silencing ,RNA, Small Interfering ,Gene knockout ,Mice, Knockout ,Gene knockdown ,Thymosin ,Gene Expression Regulation, Developmental ,Heart ,Coronary Vessels ,Cell biology ,Thymosin beta-4 ,medicine.anatomical_structure ,Models, Animal ,Immunology ,Female ,Cardiology and Cardiovascular Medicine ,Blood vessel - Abstract
Rationale: Thymosin beta 4 (Tβ4) is a 43–amino acid factor encoded by an X-linked gene. Recent studies have suggested that Tβ4 is a key factor in cardiac development, growth, disease, epicardial integrity, and blood vessel formation. Cardiac-specific short hairpin (sh)RNA knockdown of tβ4 has been reported to result in embryonic lethality at E14.5–16.5, with severe cardiac and angiogenic defects. However, this shRNA tβ4 -knockdown model did not completely abrogate Tβ4 expression. To completely ablate Tβ4 and to rule out the possibility of off-target effects associated with shRNA gene silencing, further studies of global or cardiac-specific knockouts are critical. Objective: We examined the role of Tβ4 in developing and adult heart through global and cardiac specific t β4-knockout mouse models. Methods and Results: Global t β 4 -knockout mice were born at mendelian ratios and exhibited normal heart and blood vessel formation. Furthermore, in adult global t β 4 -knockout mice, cardiac function, capillary density, expression of key cardiac fetal and angiogenic genes, epicardial marker expression, and extracellular matrix deposition were indistinguishable from that of controls. Tissue-specific t β 4 -deficient mice, generated by crossing t β 4 -floxed mice to Nkx2.5-Cre and α MHC-Cre , were also found to have no phenotype. Conclusions: We conclude that Tβ4 is dispensable for embryonic viability, heart development, coronary vessel development, and adult myocardial function.
- Published
- 2012
20. MTORC1 regulates cardiac function and myocyte survival through 4E-BP1 inhibition in mice
- Author
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Denghong, Zhang, Riccardo, Contu, Michael V G, Latronico, Jianlin, Zhang, Jian Ling, Zhang, Roberto, Rizzi, Daniele, Catalucci, Shigeki, Miyamoto, Katherine, Huang, Marcello, Ceci, Yusu, Gu, Nancy D, Dalton, Kirk L, Peterson, Kun-Liang, Guan, Joan Heller, Brown, Ju, Chen, Nahum, Sonenberg, and Gianluigi, Condorelli
- Subjects
Cardiomyopathy, Dilated ,Male ,Cardiac function curve ,medicine.medical_specialty ,Cell Survival ,Apoptosis ,Cardiomegaly ,Cell Cycle Proteins ,mTORC1 ,Mechanistic Target of Rapamycin Complex 1 ,Mice ,Clinical investigation ,Internal medicine ,Eukaryotic initiation factor ,Animals ,Myocyte ,Medicine ,Initiation factor ,Myocytes, Cardiac ,Eukaryotic Initiation Factors ,Phosphorylation ,Mechanistic target of rapamycin ,PI3K/AKT/mTOR pathway ,Adaptor Proteins, Signal Transducing ,Mice, Knockout ,Pressure overload ,biology ,business.industry ,Cell growth ,Myocardium ,TOR Serine-Threonine Kinases ,Autophagy ,Proteins ,Heart ,General Medicine ,Phosphoproteins ,Cell biology ,Mice, Inbred C57BL ,Endocrinology ,Multiprotein Complexes ,Cardiology ,biology.protein ,Female ,Carrier Proteins ,Corrigendum ,business ,Transcription Factors ,Research Article - Abstract
Mechanistic target of rapamycin (MTOR) plays a critical role in the regulation of cell growth and in the response to energy state changes. Drugs inhibiting MTOR are increasingly used in antineoplastic therapies. Myocardial MTOR activity changes during hypertrophy and heart failure (HF). However, whether MTOR exerts a positive or a negative effect on myocardial function remains to be fully elucidated. Here, we show that ablation of Mtor in the adult mouse myocardium results in a fatal, dilated cardiomyopathy that is characterized by apoptosis, autophagy, altered mitochondrial structure, and accumulation of eukaryotic translation initiation factor 4E–binding protein 1 (4E-BP1). 4E-BP1 is an MTOR-containing multiprotein complex-1 (MTORC1) substrate that inhibits translation initiation. When subjected to pressure overload, Mtor-ablated mice demonstrated an impaired hypertrophic response and accelerated HF progression. When the gene encoding 4E-BP1 was ablated together with Mtor, marked improvements were observed in apoptosis, heart function, and survival. Our results demonstrate a role for the MTORC1 signaling network in the myocardial response to stress. In particular, they highlight the role of 4E-BP1 in regulating cardiomyocyte viability and in HF. Because the effects of reduced MTOR activity were mediated through increased 4E-BP1 inhibitory activity, blunting this mechanism may represent a novel therapeutic strategy for improving cardiac function in clinical HF.
- Published
- 2010
21. Reduced thin filament length in nebulin-knockout skeletal muscle alters isometric contractile properties
- Author
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Marie Louise Bang, Jianlin Zhang, Ju Chen, David S. Gokhin, and Richard L. Lieber
- Subjects
Male ,Functional role ,medicine.medical_specialty ,Physiology ,Proteolipids ,Muscle Proteins ,Muscle Cell Biology and Cell Motility ,Isometric exercise ,Myopathies, Nemaline ,Models, Biological ,Mice ,Nebulin ,Nemaline myopathy ,Isometric Contraction ,Tensile Strength ,Internal medicine ,Myosin ,medicine ,Animals ,Muscle, Skeletal ,Actin ,Mice, Knockout ,Myosin Heavy Chains ,biology ,Skeletal muscle ,Cell Biology ,medicine.disease ,Actin cytoskeleton ,Cell biology ,Actin Cytoskeleton ,Endocrinology ,medicine.anatomical_structure ,biology.protein ,Female - Abstract
Nebulin (NEB) is a large, rod-like protein believed to dictate actin thin filament length in skeletal muscle. NEB gene defects are associated with congenital nemaline myopathy. The functional role of NEB was investigated in gastrocnemius muscles from neonatal wild-type (WT) and NEB knockout (NEB-KO) mice, whose thin filaments have uniformly shorter lengths compared with WT mice. Isometric stress production in NEB-KO skeletal muscle was reduced by 27% compared with WT skeletal muscle on postnatal day 1 and by 92% on postnatal day 7, consistent with functionally severe myopathy. NEB-KO muscle was also more susceptible to a decline in stress production during a bout of 10 cyclic isometric tetani. Length-tension properties in NEB-KO muscle were altered in a manner consistent with reduced thin filament length, with length-tension curves from NEB-KO muscle demonstrating a 7.4% narrower functional range and an optimal length reduced by 0.13 muscle lengths. Expression patterns of myosin heavy chain isoforms and total myosin content did not account for the functional differences between WT and NEB-KO muscle. These data indicate that NEB is essential for active stress production, maintenance of functional integrity during cyclic activation, and length-tension properties consistent with a role in specifying normal thin filament length. Continued analysis of NEB's functional properties will strengthen the understanding of force transmission and thin filament length regulation in skeletal muscle and may provide insights into the molecular processes that give rise to nemaline myopathy.
- Published
- 2009
22. α3 Integrin of Cell-Cell Contact Mediates Kidney Fibrosis by Integrin-Linked Kinase in Proximal Tubular E-Cadherin Deficient Mice
- Author
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Yuan M. Wang, Ye Zhao, Min Pang, Hong Zhao, Yiping Wang, So R. Lee, Isabella Rubera, Qi Cao, Ya Wang, Jianlin Zhang, Thian Kui Tan, Vincent W. Lee, Xinrui Tian, Michel Tauc, David Harris, Tzu-Ting Hsu, Xi Qiao, J. Guy Lyons, Changqi Wang, Stephen I. Alexander, Patrick P.L. Tam, Guoping Zheng, Hai-Long Wang, David A.F. Loebel, Shandong University, Key Laboratory of Functional Inorganic Material Chemistry (KLFIMC), Université de Heilongjiang, China Agricultural University (CAU), Key Laboratory of Resource Biology and Biotechnology, Northwest University, Xi'an, Laboratoire CNRS 3093, Université de Nice-Sophia Antipolis, Centre National de la Recherche Scientifique (CNRS), Key Laboratory of Intelligent Information Processing, Institute of Computing Technology [Beijing], Chinese Academy of Sciences [Changchun Branch] (CAS), College of Life Sciences, and Peking Univertsity
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Chromatin Immunoprecipitation ,[SDV]Life Sciences [q-bio] ,Integrin ,Blotting, Western ,Biology ,Protein Serine-Threonine Kinases ,Real-Time Polymerase Chain Reaction ,Pathology and Forensic Medicine ,CDH1 ,Kidney Tubules, Proximal ,03 medical and health sciences ,Mice ,Microscopy, Electron, Transmission ,Fibrosis ,Internal medicine ,medicine ,Cell Adhesion ,Animals ,Immunoprecipitation ,Integrin-linked kinase ,Cell adhesion ,ComputingMilieux_MISCELLANEOUS ,Mice, Knockout ,Cadherin ,Kinase ,Integrin alpha3beta1 ,medicine.disease ,Cadherins ,Immunohistochemistry ,Cell biology ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,biology.protein ,Kidney Diseases ,Proto-oncogene tyrosine-protein kinase Src ,Signal Transduction - Abstract
Loss of E-cadherin marks a defect in epithelial integrity and polarity during tissue injury and fibrosis. Whether loss of E-cadherin plays a causal role in fibrosis is uncertain. α3β1 Integrin has been identified to complex with E-cadherin in cell-cell adhesion, but little is known about the details of their cross talk. Herein, E-cadherin gene ( Cdh1 ) was selectively deleted from proximal tubules of murine kidney by Sglt2Cre . Ablation of E-cadherin up-regulated α3β1 integrin at cell-cell adhesion. E-cadherin–deficient proximal tubular epithelial cell displayed enhanced transforming growth factor-β1–induced α-smooth muscle actin (α-SMA) and vimentin expression, which was suppressed by siRNA silencing of α3 integrin, but not β1 integrin. Up-regulation of transforming growth factor-β1–induced α-SMA was mediated by an α3 integrin-dependent increase in integrin-linked kinase (ILK). Src phosphorylation of β-catenin and consequent p-β-catenin-Y654/p-Smad2 transcriptional complex underlies the transcriptional up-regulation of ILK. Kidney fibrosis after unilateral ureteric obstruction or ischemia reperfusion was increased in proximal tubule E-cadherin–deficient mice in comparison to that of E-cadherin intact control mice. The exacerbation of fibrosis was explained by the α3 integrin-dependent increase of ILK, β-catenin nuclear translocation, and α-SMA/proximal tubular–specific Cre double positive staining in proximal tubular epithelial cell. These studies delineate a nonconventional integrin/ILK signaling by α3 integrin–dependent Src/p-β-catenin-Y654/p-Smad2–mediated up-regulation of ILK through which loss of E-cadherin leads to kidney fibrosis.
- Published
- 2015
23. Cypher and Enigma homolog protein are essential for cardiac development and embryonic survival
- Author
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Stephan Lange, Hongqiang Cheng, Yongxin Mu, Angela K. Peter, Jennifer Veevers, Lizhu Lin, Xi Fang, Xinmin Zhou, Sylvia M. Evans, Jianlin Zhang, Ju Chen, Kunfu Ouyang, and Ran Jing
- Subjects
Muscle Proteins ,Z-line ,030204 cardiovascular system & hematology ,Cardiorespiratory Medicine and Haematology ,Bioinformatics ,Cardiovascular ,Sarcomere ,Mice ,0302 clinical medicine ,Risk Factors ,Medicine ,Protein Isoforms ,030212 general & internal medicine ,Z‐line ,Original Research ,Mice, Knockout ,0303 health sciences ,Embryonic heart ,embryonic lethality ,Microfilament Proteins ,Adaptor Proteins ,Embryo ,LIM Domain Proteins ,Cell biology ,Heart Disease ,Cardiology ,Muscle ,medicine.symptom ,Cardiology and Cardiovascular Medicine ,Cardiomyopathies ,Muscle contraction ,Gene isoform ,medicine.medical_specialty ,Multiprotein complex ,Knockout ,1.1 Normal biological development and functioning ,Embryonic Development ,Cypher ,Enigma Homolog ,Striated ,03 medical and health sciences ,Internal medicine ,Genetics ,Animals ,030304 developmental biology ,Adaptor Proteins, Signal Transducing ,business.industry ,Myocardium ,Embryogenesis ,Signal Transducing ,Correction ,Embryonic stem cell ,Muscle, Striated ,Enigma homolog protein ,business ,030217 neurology & neurosurgery - Abstract
Background The striated muscle Z‐line, a multiprotein complex at the boundary between sarcomeres, plays an integral role in maintaining striated muscle structure and function. Multiple Z‐line‐associated proteins have been identified and shown to play an increasingly important role in the pathogenesis of human cardiomyopathy. Cypher and its close homologue, Enigma homolog protein ( ENH ), are 2 Z‐line proteins previously shown to be individually essential for maintenance of postnatal cardiac function and stability of the Z‐line during muscle contraction, but dispensable for cardiac myofibrillogenesis and development. Methods and Results The current studies were designed to test whether Cypher and ENH play redundant roles during embryonic development. Here, we demonstrated that mice lacking both ENH and Cypher exhibited embryonic lethality and growth retardation. Lethality in double knockout embryos was associated with cardiac dilation and abnormal Z‐line structure. In addition, when ENH was ablated in conjunction with selective ablation of either Cypher short isoforms (CypherS), or Cypher long isoforms (CypherL), only the latter resulted in embryonic lethality. Conclusions Cypher and ENH redundantly play an essential role in sustaining Z‐line structure from the earliest stages of cardiac function, and are redundantly required to maintain normal embryonic heart function and embryonic viability.
- Published
- 2015
24. Normalization of Naxos plakoglobin levels restores cardiac function in mice
- Author
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Yusu Gu, Hongqiang Cheng, Xi Fang, Xinmin Zhou, William H. Bradford, Yongxin Mu, Kunfu Ouyang, Matthew J. Stroud, Kirk L. Peterson, Kensuke Kimura, Ju Chen, Zhiwei Zhang, Nancy D. Dalton, and Jianlin Zhang
- Subjects
RNA Stability ,Nonsense-mediated decay ,Messenger ,Arrhythmias ,Lethal ,Cardiovascular ,Medical and Health Sciences ,Mice ,Keratoderma, Palmoplantar ,2.1 Biological and endogenous factors ,Myocytes, Cardiac ,Gene Knock-In Techniques ,Aetiology ,Frameshift Mutation ,Wnt Signaling Pathway ,Arrhythmogenic Right Ventricular Dysplasia ,Sequence Deletion ,Brief Report ,Wnt signaling pathway ,Palmoplantar ,General Medicine ,Phenotype ,Arrhythmogenic right ventricular dysplasia ,Cell biology ,medicine.anatomical_structure ,Heart Disease ,Codon, Nonsense ,Intercalated disc ,Cardiac ,medicine.medical_specialty ,Heart Ventricles ,Immunology ,Plakoglobin ,Biology ,Frameshift mutation ,Naxos disease ,Rare Diseases ,Internal medicine ,medicine ,Genetics ,Animals ,Humans ,RNA, Messenger ,Codon ,Myocytes ,Myocardium ,Arrhythmias, Cardiac ,medicine.disease ,Fibrosis ,Myocardial Contraction ,Peptide Fragments ,Nonsense Mediated mRNA Decay ,Keratoderma ,Endocrinology ,Nonsense ,Genes ,Desmoplakins ,RNA ,Genes, Lethal ,gamma Catenin ,Hair Diseases - Abstract
Arrhythmogenic cardiomyopathy (AC) is associated with mutations in genes encoding intercalated disc proteins and ultimately results in sudden cardiac death. A subset of patients with AC have the autosomal recessive cardiocutaneous disorder Naxos disease, which is caused by a 2–base pair deletion in the plakoglobin-encoding gene JUP that results in a truncated protein with reduced expression. In mice, cardiomyocyte-specific plakoglobin deficiency recapitulates many aspects of human AC, and overexpression of the truncated Naxos-associated plakoglobin also results in an AC-like phenotype; therefore, it is unclear whether Naxos disease results from loss or gain of function consequent to the plakoglobin mutation. Here, we generated 2 knockin mouse models in which endogenous Jup was engineered to express the Naxos-associated form of plakoglobin. In one model, Naxos plakoglobin bypassed the nonsense-mediated mRNA decay pathway, resulting in normal levels of the truncated plakoglobin. Moreover, restoration of Naxos plakoglobin to WT levels resulted in normal heart function. Together, these data indicate that a gain of function in the truncated form of the protein does not underlie the clinical phenotype of patients with Naxos disease and instead suggest that insufficiency of the truncated Naxos plakoglobin accounts for disease manifestation. Moreover, these results suggest that increasing levels of truncated or WT plakoglobin has potential as a therapeutic approach to Naxos disease.
- Published
- 2015
25. Disruption of both nesprin 1 and desmin results in nuclear anchorage defects and fibrosis in skeletal muscle
- Author
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Ju Chen, Ling T. Guo, Richard L. Lieber, Jianlin Zhang, Kunfu Ouyang, Zhiwei Zhang, G. Diane Shelton, Mark A. Chapman, and Indroneal Banerjee
- Subjects
Male ,medicine.medical_specialty ,Knockout ,1.1 Normal biological development and functioning ,Nerve Tissue Proteins ,Biology ,Medical and Health Sciences ,Muscular Dystrophies ,Desmin ,Extracellular matrix ,Mice ,Underpinning research ,Internal medicine ,Genetics ,medicine ,Animals ,Humans ,2.1 Biological and endogenous factors ,Nuclear protein ,Mechanotransduction ,Aetiology ,Cytoskeleton ,Muscle, Skeletal ,Molecular Biology ,Genetics (clinical) ,Mice, Knockout ,Cell Nucleus ,Genetics & Heredity ,Nesprin ,Skeletal muscle ,Nuclear Proteins ,General Medicine ,Articles ,Skeletal ,Biological Sciences ,Fibrosis ,Cell biology ,Cell nucleus ,Cytoskeletal Proteins ,Protein Transport ,medicine.anatomical_structure ,Endocrinology ,Musculoskeletal ,Muscle ,Female - Abstract
Proper localization and anchorage of nuclei within skeletal muscle is critical for cellular function. Alterations in nuclear anchoring proteins modify a number of cellular functions including mechanotransduction, nuclear localization, chromatin positioning/compaction and overall organ function. In skeletal muscle, nesprin 1 and desmin are thought to link the nucleus to the cytoskeletal network. Thus, we hypothesize that both of these factors play a key role in skeletal muscle function. To examine this question, we utilized global ablation murine models of nesprin 1, desmin or both nesprin 1 and desmin. Herein, we have created the nesprin-desmin double-knockout (DKO) mouse, eliminating a major fraction of nuclear-cytoskeletal connections and enabling understanding of the importance of nuclear anchorage in skeletal muscle. Globally, DKO mice are marked by decreased lifespan, body weight and muscle strength. With regard to skeletal muscle, DKO myonuclear anchorage was dramatically decreased compared with wild-type, nesprin 1(-/-) and desmin(-/-) mice. Additionally, nuclear-cytoskeletal strain transmission was decreased in DKO skeletal muscle. Finally, loss of nuclear anchorage in DKO mice coincided with a fibrotic response as indicated by increased collagen and extracellular matrix deposition and increased passive mechanical properties of muscle bundles. Overall, our data demonstrate that nesprin 1 and desmin serve redundant roles in nuclear anchorage and that the loss of nuclear anchorage in skeletal muscle results in a pathological response characterized by increased tissue fibrosis and mechanical stiffness.
- Published
- 2014
26. The ryanodine receptor store-sensing gate controls Ca2+ waves and Ca2+-triggered arrhythmias
- Author
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Jingqun Zhang, Lin Zhang, Jeff Bolstad, Yunlong Bai, Henry J. Duff, Megan L. O'Mara, Tao Mi, Qiang Zhou, Anne M. Gillis, D. Peter Tieleman, Huihui Kong, Ju Chen, Xiaowei Zhong, Peter P. Jones, Cuihong Xie, Michael Fill, Long-Sheng Song, Wenqian Chen, Yingjie Liu, Xixi Tian, Ang Guo, Ruiwu Wang, Hongqiang Cheng, Lisa Semeniuk, S. R. Wayne Chen, Jianlin Zhang, and Biyi Chen
- Subjects
Patch-Clamp Techniques ,Lipid Bilayers ,Arrhythmias ,Cardiovascular ,Ryanodine receptor 2 ,Medical and Health Sciences ,chemistry.chemical_compound ,Mice ,Myocyte ,Site-Directed ,2.1 Biological and endogenous factors ,Myocytes, Cardiac ,Inositol ,Gene Knock-In Techniques ,Aetiology ,Microscopy ,Microscopy, Confocal ,Ryanodine receptor ,Cardiac muscle ,General Medicine ,musculoskeletal system ,Cell biology ,medicine.anatomical_structure ,Heart Disease ,Biochemistry ,Echocardiography ,Confocal ,cardiovascular system ,tissues ,Cardiac ,Intracellular ,Immunoblotting ,Immunology ,Biology ,Sudden death ,Article ,General Biochemistry, Genetics and Molecular Biology ,Caffeine ,medicine ,Animals ,Humans ,Point Mutation ,Patch clamp ,DNA Primers ,Myocytes ,Arrhythmias, Cardiac ,Ryanodine Receptor Calcium Release Channel ,HEK293 Cells ,chemistry ,Mutagenesis ,Mutagenesis, Site-Directed ,Calcium - Abstract
Spontaneous Ca(2+) release from intracellular stores is important for various physiological and pathological processes. In cardiac muscle cells, spontaneous store overload-induced Ca(2+) release (SOICR) can result in Ca(2+) waves, a major cause of ventricular tachyarrhythmias (VTs) and sudden death. The molecular mechanism underlying SOICR has been a mystery for decades. Here we show that a point mutation, E4872A, in the helix bundle crossing region (the proposed gate) of the cardiac ryanodine receptor (RyR2) completely abolishes luminal, but not cytosolic, Ca(2+) activation of RyR2. The introduction of metal-binding histidines at this site converts RyR2 into a luminal Ni(2+)-gated channel. Mouse hearts harboring a heterozygous RyR2 mutation at this site (E4872Q) are resistant to SOICR and are completely protected against Ca(2+)-triggered VTs. These data show that the RyR2 gate directly senses luminal (store) Ca(2+), explaining the regulation of RyR2 by luminal Ca(2+), the initiation of Ca(2+) waves and Ca(2+)-triggered arrhythmias. This newly identified store-sensing gate structure is conserved in all RyR and inositol 1,4,5-trisphosphate receptor isoforms.
- Published
- 2014
27. Neuronal Death and Perinatal Lethality in Voltage-Gated Sodium Channel αII-Deficient Mice
- Author
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Marco Caprini, Cornelis Murre, Mauricio Montal, Eliezer Masliah, Rosa Planells-Cases, Jianlin Zhang, R.R. Rivera, and Edward Rockenstein
- Subjects
Programmed cell death ,Restriction Mapping ,Biophysics ,Hippocampus ,Apoptosis ,Neocortex ,Biology ,Sodium Channels ,Mice ,medicine ,Animals ,Fetal Death ,Cells, Cultured ,Mice, Knockout ,Neurons ,Recombination, Genetic ,Cell Death ,Sodium channel ,Embryogenesis ,Brain ,Cell biology ,medicine.anatomical_structure ,Animals, Newborn ,Immunology ,Knockout mouse ,Brainstem ,Brain Stem ,Saxitoxin ,Research Article - Abstract
Neural activity is crucial for cell survival and fine patterning of neuronal connectivity during neurodevelopment. To investigate the role in vivo of sodium channels (NaCh) in these processes, we generated knockout mice deficient in brain NaChalpha(II). NaChalpha(II)(-/-) mice were morphologically and organogenically indistinguishable from their NaChalpha(+/-) littermates. Notwithstanding, NaChalpha(II)(-/-) mice died perinatally with severe hypoxia and massive neuronal apoptosis, notably in the brainstem. Sodium channel currents recorded from cultured neurons of NaChalpha(II)(-/-) mice were sharply attenuated. Death appears to arise from severe hypoxia consequent to the brainstem deficiency of NaChalpha(II). NaChalpha(II) expression is, therefore, redundant for embryonic development but essential for postnatal survival.
- Published
- 2000
28. The nebulin SH3 domain is dispensable for normal skeletal muscle structure but is required for effective active load bearing in mouse
- Author
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Kazunori Takano, Carmen Vitiello, Paolo Kunderfranco, Gerald Coulis, Serena Camerini, Ju Chen, Marie Louise Bang, Jianlin Zhang, Pradeep K. Luther, Daniel L. Yamamoto, Marco Crescenzi, Alessandra Castaldi, Takeshi Endo, Richard L. Lieber, Fabio Piaser, Peter J. Eggenhuizen, Maria Carmela Filomena, and David S. Gokhin
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Male ,Gene Expression ,Muscle Proteins ,Wiskott-Aldrich Syndrome Protein, Neuronal ,macromolecular substances ,Biology ,Myopathies, Nemaline ,Sarcomere ,SH3 domain ,Zyxin ,Weight-Bearing ,Nebulin ,Mice ,Nemaline myopathy ,Elastic Modulus ,Isometric Contraction ,Tensile Strength ,medicine ,Animals ,Humans ,Muscle, Skeletal ,Molecular Biology ,Actin ,Excitation Contraction Coupling ,Genetics ,Palladin ,Skeletal muscle ,Cell Biology ,Anatomy ,medicine.disease ,Phosphoproteins ,Congenital myopathy ,Cell biology ,Protein Structure, Tertiary ,Cytoskeletal Proteins ,medicine.anatomical_structure ,biology.protein ,Female ,Developmental Biology ,Protein Binding ,Research Article - Abstract
Nemaline myopathy (NM) is a congenital myopathy with an estimated incidence of 1∶50,000 live births. It is caused by mutations in thin filament components, including nebulin, which accounts for about 50% of the cases. The identification of NM cases with nonsense mutations resulting in loss of the extreme C-terminal SH3 domain of nebulin suggests an important role of the nebulin SH3 domain, which is further supported by the recent demonstration of its role in IGF-1-induced sarcomeric actin filament formation through targeting of N-WASP to the Z-line. To provide further insights into the functional significance of the nebulin SH3 domain in the Z-disk and to understand the mechanisms by which truncations of nebulin lead to NM, we took two approaches: (1) an affinity-based proteomic screening to identify novel interaction partners of the nebulin SH3 domain; and (2) generation and characterization of a novel knockin mouse model with a premature stop codon in the nebulin gene, eliminating its C-terminal SH3 domain (NebΔSH3 mouse). Surprisingly, detailed analyses of NebΔSH3 mice revealed no structural or histological skeletal muscle abnormalities and no changes in gene expression or localization of interaction partners of the nebulin SH3 domain, including myopalladin, palladin, zyxin and N-WASP. Also, no significant effect on peak isometric stress production, passive tensile stress or Young's modulus was found. However, NebΔSH3 muscle displayed a slightly altered force–frequency relationship and was significantly more susceptible to eccentric contraction-induced injury, suggesting that the nebulin SH3 domain protects against eccentric contraction-induced injury and possibly plays a role in fine-tuning the excitation–contraction coupling mechanism.
- Published
- 2013
29. Matrix metalloproteinase 9-dependent Notch signaling contributes to kidney fibrosis through peritubular endothelial–mesenchymal transition
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Lixin Liu, Stephen I. Alexander, Vincent W. Lee, Qi Cao, Lihua Wang, Yuan Min Wang, Ye Zhao, Hong Zhao, Yiping Wang, Ya Wang, Jianlin Zhang, David Harris, Thian Kui Tan, Xi Qiao, Yun Zhang, and Guoping Zheng
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Male ,0301 basic medicine ,CD31 ,Notch ,Endothelium ,kidney fibrosis ,Matrix metalloproteinase 9 ,Notch signaling pathway ,Vimentin ,Mesoderm ,Transforming Growth Factor beta1 ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Fibrosis ,medicine ,Animals ,Humans ,Mice, Knockout ,peritubular endothelial cells ,Mice, Inbred BALB C ,Transplantation ,Kidney ,Receptors, Notch ,biology ,Cadherin ,business.industry ,Transforming growth factor beta ,endothelial–mesenchymal transition ,medicine.disease ,3. Good health ,Basic Research ,030104 developmental biology ,medicine.anatomical_structure ,Nephrology ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,Kidney Diseases ,ORIGINAL ARTICLES ,business ,Signal Transduction ,Ureteral Obstruction - Abstract
Background: Endothelial cells are known to contribute to kidney fibrosis via endothelial–mesenchymal transition (EndoMT). Matrix metalloproteinase 9 (MMP-9) is known to be profibrotic. However, whether MMP-9 contributes to kidney fibrosis via EndoMT is unknown. Methods: Primary mouse renal peritubular endothelial cells (MRPECs) were isolated and treated by recombinant human transforming growth factor beta 1 (rhTGF-β1) with or without MMP-9 inhibitor or by recombinant human MMP-9 (rhMMP-9) alone. Kidney fibrosis was induced by unilateral ureteral obstruction (UUO) in MMP-9 knockout (KO) and wide-type (WT) control mice. The effects of MMP-9 on EndoMT of MRPECs and kidney fibrosis were examined. Results: We showed that MRPECs underwent EndoMT after rhTGF-β1 treatment or in UUO kidney as evidenced by decreased expression of endothelial markers, vascular endothelial cadherin (VE-cadherin) and CD31, and increased levels of mesenchymal markers, α-smooth muscle actin (α-SMA) and vimentin. The expression of fibrosis markers was also up-regulated significantly after rhTGF-β1 treatment in MRPECs. The EndoMT and fibrosis markers were significantly less in rhTGF-β1-treated MMP-9 KO MRPECs, whereas MMP-9 alone was sufficient to induce EndoMT in MRPECs. UUO kidney of MMP-9 KO mice showed significantly less interstitial fibrosis and EndoMT in MRPECs. Notch signaling shown by Notch intracellular domain (NICD) was increased, while Notch-1 was decreased in rhTGF-β1-treated MRPECs of MMP-9 WT but not MMP-9 KO mice. Inhibition of MMP-9 or Notch signaling prevented rhTGF-β1- or rhMMP-9-induced α-SMA and NICD upregulation in MRPECs. UUO kidney of MMP-9 KO mice had less staining of Notch signaling transcription factor Hey-1 in VE-cadherin-positive MRPECs than WT controls. Conclusions: Our results demonstrate that MMP-9-dependent Notch signaling plays an important role in kidney fibrosis through EndoMT of MRPECs.
- Published
- 2016
30. Nesprin 1 is critical for nuclear positioning and anchorage
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G. Diane Shelton, Yujie Liu, Nancy D. Dalton, Ling T. Guo, Amanda Felder, Ju Chen, Andrew P. Mizisin, Stephan Lange, Kirk L. Peterson, Yusu Gu, Richard L. Lieber, and Jianlin Zhang
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Male ,Amino Acid Motifs ,Nerve Tissue Proteins ,Biology ,Mice ,Genetics ,medicine ,Animals ,Humans ,Nuclear protein ,Muscular dystrophy ,Muscle, Skeletal ,Molecular Biology ,Genetics (clinical) ,Cell Nucleus ,Mice, Knockout ,Nesprin ,Skeletal muscle ,Nuclear Proteins ,General Medicine ,Articles ,Actin cytoskeleton ,medicine.disease ,Muscular Dystrophy, Emery-Dreifuss ,Transport protein ,Cell biology ,Mice, Inbred C57BL ,Cell nucleus ,Cytoskeletal Proteins ,Disease Models, Animal ,Protein Transport ,medicine.anatomical_structure ,Biochemistry ,Knockout mouse ,Female ,Protein Binding - Abstract
Nesprin 1 is an outer nuclear membrane protein that is thought to link the nucleus to the actin cytoskeleton. Recent data suggest that mutations in Nesprin 1 may also be involved in the pathogenesis of Emery-Dreifuss muscular dystrophy. To investigate the function of Nesprin 1 in vivo, we generated a mouse model in which all isoforms of Nesprin 1 containing the C-terminal spectrin-repeat region with or without KASH domain were ablated. Nesprin 1 knockout mice are marked by decreased survival rates, growth retardation and increased variability in body weight. Additionally, nuclear positioning and anchorage are dysfunctional in skeletal muscle from knockout mice. Physiological testing demonstrated no significant reduction in stress production in Nesprin 1-deficient skeletal muscle in either neonatal or adult mice, but a significantly lower exercise capacity in knockout mice. Nuclear deformation testing revealed ineffective strain transmission to nuclei in muscle fibers lacking Nesprin 1. Overall, our data show that Nesprin 1 is essential for normal positioning and anchorage of nuclei in skeletal muscle.
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- 2009
31. The knockout of miR-143 and -145 alters smooth muscle cell maintenance and vascular homeostasis in mice: Correlates with human disease
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Ciro Indolfi, Daniele Catalucci, Gianluigi Condorelli, Sara A. Courtneidge, Kirk L. Peterson, Leonardo Elia, Jianlin Zhang, Manuela Quintavalle, Michael V.G. Latronico, Riccardo Contu, Luca Cossu, and Ju Chen
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Pathology ,medicine.medical_specialty ,Vascular smooth muscle ,Cellular differentiation ,Knockout ,Myocytes, Smooth Muscle ,Biology ,Muscle, Smooth, Vascular ,Article ,Cell Line ,Pathogenesis ,Mice ,Smooth Muscle ,Vascular ,microRNA ,Gene expression ,medicine ,vascular smooth muscle cells ,Animals ,Humans ,Homeostasis ,Molecular Biology ,Aorta ,Cell Proliferation ,Mice, Knockout ,Myocytes ,Base Sequence ,Vascular disease ,Cell growth ,Cell Differentiation ,Cell Biology ,medicine.disease ,Cell biology ,Rats ,MicroRNAs ,cardiovascular system ,Muscle ,Smooth - Abstract
Mechanisms controlling vascular smooth muscle cell (VSMC) plasticity and renewal still remain to be completely elucidated. A class of small RNAs called microRNAs (miRs) regulate gene expression at the post-transcriptional level. Here we demonstrate a critical role of the miR-143/145 cluster in SMC differentiation and vascular pathogenesis, also through the generation of a mouse model of miR-143 and -145 knockout. We determined that the expression of miR-143 and -145 is decreased in acute and chronic vascular stress (transverse aortic constriction and in aortas of the ApoE knockout mouse). In human aortic aneurysms, the expression of miR-143 and -145 was significantly decreased compared to control aortas. In addition, overexpression of miR-143 and -145 decreased neointimal formation in a rat model of acute vascular injury. An in-depth analysis of the miR-143/145 knockout mouse model demonstrated that this miR cluster is expressed mostly in the SMC compartment, both during development and post-natally, in vessels and SMC-containing organs. Loss of miR-143 and miR-145 expression induces structural modifications of the aorta, due to an incomplete differentiation of VSMCs. In conclusion, our results demonstrate that the miR-143/145 gene cluster plays a critical role during SMC differentiation and strongly suggest its involvement in the reversion of the VSMC differentiation phenotype that occurs during vascular disease.
- Published
- 2009
32. Cardiac-specific ablation of Cypher leads to a severe form of dilated cardiomyopathy with premature death
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Kirk L. Peterson, Liang Han, Li Cui, Hongqiang Cheng, Ju Chen, Nancy D. Dalton, Kunfu Ouyang, Ting Zhao, Jianlin Zhang, Ming Zheng, Yusu Gu, Xiaodong Li, and Marie Louise Bang
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Cardiac function curve ,Cardiomyopathy, Dilated ,Heart disease ,Cardiomyopathy ,Muscle Proteins ,Biology ,Cell Line ,Mice ,Genetics ,medicine ,Myotilin ,Animals ,Humans ,Mortality ,Molecular Biology ,Genetics (clinical) ,Adaptor Proteins, Signal Transducing ,Homeodomain Proteins ,Mice, Knockout ,Myocardium ,Microfilament Proteins ,Hypertrophic cardiomyopathy ,Dilated cardiomyopathy ,Heart ,General Medicine ,Articles ,LIM Domain Proteins ,medicine.disease ,Phenotype ,Disease Models, Animal ,Heart failure ,Mutation ,Cancer research ,cardiovascular system ,Carrier Proteins ,Protein Binding - Abstract
Accumulating data suggest a link between alterations/deficiencies in cytoskeletal proteins and the progression of cardiomyopathy and heart failure, although the molecular basis for this link remains unclear. Cypher/ZASP is a cytoskeletal protein localized in the sarcomeric Z-line. Mutations in its encoding gene have been identified in patients with isolated non-compaction of the left ventricular myocardium, dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy. To explore the role of Cypher in myocardium and to better understand molecular mechanisms by which mutations in cypher cause cardiomyopathy, we utilized a conditional approach to knockout Cypher, specially in either developing or adult myocardium. Cardiac-specific Cypher knockout (CKO) mice developed a severe form of DCM with disrupted cardiomyocyte ultrastructure and decreased cardiac function, which eventually led to death before 23 weeks of age. A similar phenotype was observed in inducible cardiac-specific CKO mice in which Cypher was specifically ablated in adult myocardium. In both cardiac-specific CKO models, ERK and Stat3 signaling pathways were augmented. Finally, we demonstrate the specific binding of Cypher's PDZ domain to the C-terminal region of both calsarcin-1 and myotilin within the Z-line. In conclusion, our studies suggest that (i) Cypher plays a pivotal role in maintaining adult cardiac structure and cardiac function through protein-protein interactions with other Z-line proteins, (ii) myocardial ablation of Cypher results in DCM with premature death and (iii) specific signaling pathways participate in Cypher mutant-mediated dysfunction of the heart, and may in concert facilitate the progression to heart failure.
- Published
- 2008
33. Syncoilin is required for generating maximum isometric stress in skeletal muscle but dispensable for muscle cytoarchitecture
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Richard L. Lieber, Li Cui, Jianlin Zhang, Kirk L. Peterson, Nancy D. Dalton, Maria Cecilia Scimia, Ju Chen, David S. Gokhin, Yingchun Lu, Yusu Gu, Xiaodong Li, and Marie Louise Bang
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medicine.medical_specialty ,Physiology ,Muscle Fibers, Skeletal ,Restriction Mapping ,Muscle Proteins ,Isometric exercise ,Biology ,Sarcomere ,Ventricular Function, Left ,Article ,Desmin ,Mice ,Intermediate Filament Proteins ,Internal medicine ,medicine ,Animals ,Intermediate filament ,Muscle, Skeletal ,DNA Primers ,Mice, Knockout ,Sarcolemma ,Hemodynamics ,Skeletal muscle ,Cell Biology ,Syncoilin ,Cell biology ,Hindlimb ,Endocrinology ,medicine.anatomical_structure ,Stress, Mechanical ,medicine.symptom ,Muscle contraction ,Muscle Contraction - Abstract
Syncoilin is a striated muscle-specific intermediate filament-like protein, which is part of the dystrophin-associated protein complex (DPC) at the sarcolemma and provides a link between the extracellular matrix and the cytoskeleton through its interaction with α-dystrobrevin and desmin. Its upregulation in various neuromuscular diseases suggests that syncoilin may play a role in human myopathies. To study the functional role of syncoilin in cardiac and skeletal muscle in vivo, we generated syncoilin-deficient ( syncoilin−/−) mice. Our detailed analysis of these mice up to 2 yr of age revealed that syncoilin is entirely dispensable for cardiac and skeletal muscle development and maintenance of cellular structure but is required for efficient lateral force transmission during skeletal muscle contraction. Notably, syncoilin−/−skeletal muscle generates less maximal isometric stress than wild-type (WT) muscle but is as equally susceptible to eccentric contraction-induced injury as WT muscle. This suggests that syncoilin may play a supportive role for desmin in the efficient coupling of mechanical stress between the myofibril and fiber exterior. It is possible that the reduction in isometric stress production may predispose the syncoilin skeletal muscle to a dystrophic condition.
- Published
- 2008
34. Liver X receptor activation induces apoptosis of melanoma cell through caspase pathway.
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Wenjun Zhang, Hua Jiang, Jianlin Zhang, Yinfan Zhang, Antang Liu, Yaozhong Zhao, Xiaohai Zhu, Zihao Lin, and Xiangbin Yuan
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NUCLEAR receptors (Biochemistry) ,APOPTOSIS ,CANCER cells ,LIPID metabolism ,TUMOR growth ,WESTERN immunoblotting ,MICE ,ANIMAL models in research - Abstract
Liver X receptors (LXRs) are nuclear receptors that function as ligand-activated transcription factors regulating lipid metabolism and inflammation. Recent discoveries found LXRs could regulate tumor growth in a variety of cancer cell lines. In this study, we investigated the effect of LXR activation on melanoma cell proliferation and apoptosis both in vitro and in vivo. Treatment of B16F10 and A-375 melanoma cells with synthetic LXR agonist T0901317 significantly inhibited the proliferation of melanoma cells in vitro. Meanwhile, T0901317 induced the apoptosis of B16F10 melanoma cells in a dose-dependent manner. Furthermore, western blot assay showed that the pro-apoptotic effect of T0901317 on B16F10 melanoma cells was mediated through caspase-3 pathway. Oral administration of T0901317 inhibited the growth of B16F10 melanoma in C56BL/6 mice. Altogether, this study demonstrates the critical role of LXRs in the regulation of melanoma growth and presents the LXR agonist T0901317 as a potential anti-melanoma agent. [ABSTRACT FROM AUTHOR]
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- 2014
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- View/download PDF
35. Myopalladin knockout mice develop cardiac dilation and show a maladaptive response to mechanical pressure overload.
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Filomena, Maria Carmela, Yamamoto, Daniel L., Carullo, Pierluigi, Medvedev, Roman, Ghisleni, Andrea, Piroddi, Nicoletta, Scellini, Beatrice, Crispino, Roberta, D'Autilia, Francesca, Jianlin Zhang, Felicetta, Arianna, Nemska, Simona, Serio, Simone, Tesi, Chiara, Catalucci, Daniele, Linke, Wolfgang A., Polishchuk, Roman, Poggesi, Corrado, Gautel, Mathias, and Bang, Marie-Louise
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KNOCKOUT mice , *GENETIC mutation , *GENE expression , *CONNECTIN , *THERMOPHORESIS , *MICE - Abstract
Myopalladin (MYPN) is a striated muscle-specific immunoglobulin domain-containing protein located in the sarcomeric Z-line and I-band. MYPN gene mutations are causative for dilated (DCM), hypertrophic, and restrictive cardiomyopathy. In a yeast two-hybrid screening, MYPN was found to bind to titin in the Z-line, which was confirmed by microscale thermophoresis. Cardiac analyses of MYPN knockout (MKO) mice showed the development of mild cardiac dilation and systolic dysfunction, associated with decreased myofibrillar isometric tension generation and increased resting tension at longer sarcomere lengths. MKO mice exhibited a normal hypertrophic response to transaortic constriction (TAC), but rapidly developed severe cardiac dilation and systolic dysfunction, associated with fibrosis, increased fetal gene expression, higher intercalated disc fold amplitude, decreased calsequestrin-2 protein levels, and increased desmoplakin and SORBS2 protein levels. Cardiomyocyte analyses showed delayed Ca2+ release and reuptake in unstressed MKO mice as well as reduced Ca2+ spark amplitude post-TAC, suggesting that altered Ca2+ handling may contribute to the development of DCM in MKO mice. [ABSTRACT FROM AUTHOR]
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- 2021
- Full Text
- View/download PDF
36. PTPMT1 Is Required for Embryonic Cardiac Cardiolipin Biosynthesis to Regulate Mitochondrial Morphogenesis and Heart Development.
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Ze'e Chen, Siting Zhu, Hong Wang, Li Wang, Jianlin Zhang, Yusu Gu, Changming Tan, Dhanani, Mehul, Wever, Eric, Xinru Wang, Boyu Xie, Shijia Wang, Lei Huang, van Kampen, Antoine H. C., Jie Liu, Zhen Han, Patel, Hemal H., Vaz, Frédéric M., Xi Fang, and Ju Chen
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HEART development , *CARDIOLIPIN , *MITOCHONDRIA , *BIOSYNTHESIS , *MORPHOGENESIS , *KNOCKOUT mice , *ANTIPHOSPHOLIPID syndrome , *RESEARCH , *HEART , *ANIMAL experimentation , *IMMUNOHISTOCHEMISTRY , *RESEARCH methodology , *PHOSPHATASES , *FETAL development , *MEDICAL cooperation , *EVALUATION research , *COMPARATIVE studies , *GENES , *GENE expression profiling , *PHOSPHOLIPIDS , *MICE - Published
- 2021
- Full Text
- View/download PDF
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