Cardiac physiology at the cellular level: use of cultured HL-1 cardiomyocytes for studies of cardiac muscle cell structure and function

2004 ◽  
Vol 286 (3) ◽  
pp. H823-H829 ◽  
Author(s):  
Steven M. White ◽  
Phillip E. Constantin ◽  
William C. Claycomb
Keyword(s):  
Cell Line ◽  
Cell Structure ◽  
Ischemia Reperfusion ◽  
Cellular Level ◽  
Model Systems ◽  
Cardiac Physiology ◽  
Cardiac Phenotype ◽  
Pathological Conditions ◽  
And Function

HL-1 cells are currently the only cardiomyocyte cell line available that continuously divides and spontaneously contracts while maintaining a differentiated cardiac phenotype. Extensive characterization using microscopic, genetic, immunohistochemical, electrophysiological, and pharmacological techniques has demonstrated how similar HL-1 cells are to primary cardiomyocytes. In the few years that HL-1 cells have been available, they have been used in a variety of model systems designed to answer important questions regarding cardiac biology at the cellular and molecular levels. Whereas HL-1 cells have been used to study normal cardiomyocyte function with regard to signaling, electrical, metabolic, and transcriptional regulation, they have also been used to address pathological conditions such as hypoxia, hyperglycemia-hyperinsulinemia, apoptosis, and ischemia-reperfusion. The availability of an immortalized, contractile cardiac cell line has provided investigators with a tool for probing the intricacies of cardiomyocyte function. In this review, we describe the culture and characterization of HL-1 cardiomyocytes as well as various model systems that have been developed using these cells to gain a better understanding of cardiac biology at the cellular and molecular levels.

scholarly journals Impact of Aldosterone on the Failing Myocardium: Insights from Mitochondria and Adrenergic Receptors Signaling and Function

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1552
Author(s):  
Mariona Guitart-Mampel ◽  
Pedro Urquiza ◽  
Jordana I. Borges ◽  
Anastasios Lymperopoulos ◽  
Maria E. Solesio
Keyword(s):  
Mitochondrial Dysfunction ◽  
G Protein ◽  
Adrenergic Receptor ◽  
Cellular Level ◽  
Cardiac Physiology ◽  
Failing Heart ◽  
Receptor Kinases ◽  
And Function ◽  
The Impact ◽  
G Protein Coupled

The mineralocorticoid aldosterone regulates electrolyte and blood volume homeostasis, but it also adversely modulates the structure and function of the chronically failing heart, through its elevated production in chronic human post-myocardial infarction (MI) heart failure (HF). By activating the mineralocorticoid receptor (MR), a ligand-regulated transcription factor, aldosterone promotes inflammation and fibrosis of the heart, while increasing oxidative stress, ultimately induding mitochondrial dysfunction in the failing myocardium. To reduce morbidity and mortality in advanced stage HF, MR antagonist drugs, such as spironolactone and eplerenone, are used. In addition to the MR, aldosterone can bind and stimulate other receptors, such as the plasma membrane-residing G protein-coupled estrogen receptor (GPER), further complicating it signaling properties in the myocardium. Given the salient role that adrenergic receptor (ARs)—particularly βARs—play in cardiac physiology and pathology, unsurprisingly, that part of the impact of aldosterone on the failing heart is mediated by its effects on the signaling and function of these receptors. Aldosterone can significantly precipitate the well-documented derangement of cardiac AR signaling and impairment of AR function, critically underlying chronic human HF. One of the main consequences of HF in mammalian models at the cellular level is the presence of mitochondrial dysfunction. As such, preventing mitochondrial dysfunction could be a valid pharmacological target in this condition. This review summarizes the current experimental evidence for this aldosterone/AR crosstalk in both the healthy and failing heart, and the impact of mitochondrial dysfunction in HF. Recent findings from signaling studies focusing on MR and AR crosstalk via non-conventional signaling of molecules that normally terminate the signaling of ARs in the heart, i.e., the G protein-coupled receptor-kinases (GRKs), are also highlighted.

Activins, Inhibins, and Follistatins: From Endocrinology to Signaling. A Paradigm for the New Millennium

2002 ◽  
Vol 227 (9) ◽  
pp. 724-752 ◽  
Author(s):  
Corrine Welt ◽  
Yisrael Sidis ◽  
Henry Keutmann ◽  
Alan Schneyer
Keyword(s):  
Hormone Secretion ◽  
Signal Propagation ◽  
Cellular Level ◽  
Model Systems ◽  
Pituitary Hormone ◽  
New Paradigm ◽  
Activin Signaling ◽  
Physiological Relevance ◽  
Paracrine Growth Factor

It has been 70 years since the name inhibin was used to describe a gonadal factor that negatively regulated pituitary hormone secretion. The majority of this period was required to achieve purification and definitive characterization of inhibin, an event closely followed by identification and characterization of activin and follistatin (FS). In contrast, the last 15–20 years saw a virtual explosion of information regarding the biochemistry, physiology, and biosynthesis of these proteins, as well as identification of activin receptors, and a unique mechanism for FS action—the nearly irreversible binding and neutralization of activin. Many of these discoveries have been previously summarized; therefore, this review will cover the period from the mid 1990s to present, with particular emphasis on emerging themes and recent advances. As the field has matured, recent efforts have focused more on human studies, so the endocrinology of inhibin, activin, and FS in the human is summarized first. Another area receiving significant recent attention is local actions of activin and its regulation by both FS and inhibin. Because activin and FS are produced in many tissues, we chose to focus on a few particular examples with the most extensive experimental support, the pituitary and the developing follicle, although nonreproductive actions of activin and FS are also discussed. At the cellular level, it now seems that activin acts largely as an autocrine and/or paracrine growth factor, similar to other members of the transforming growh factor β superfamily. As we discuss in the next section, its actions are regulated extracellularly by both inhibin and FS. In the final section, intracellular mediators and modulators of activin signaling are reviewed in detail. Many of these are shared with other transforming growh factor β superfamily members as well as unrelated molecules, and in a number of cases, their physiological relevance to activin signal propagation remains to be elucidated. Nevertheless, taken together, recent findings suggest that it may be more appropriate to consider a new paradigm for inhibin, activin, and FS in which activin signaling is regulated extracellularly by both inhibin and FS whereas a number of intracellular proteins act to modulate cellular responses to these activin signals. It is therefore the balance between activin and all of its modulators, rather than the actions of any one component, that determines the final biological outcome. As technology and model systems become more sophisticated in the next few years, it should become possible to test this concept directly to more clearly define the role of activin, inhibin, and FS in reproductive physiology.

scholarly journals Characterization of Glial Cell Models andIn VitroManipulation of the Neuregulin1/ErbB System

2014 ◽  
Vol 2014 ◽  
pp. 1-15 ◽  
Author(s):  
Davide Pascal ◽  
Alessia Giovannelli ◽  
Sara Gnavi ◽  
Stefan Adriaan Hoyng ◽  
Fred de Winter ◽  
...  
Keyword(s):  
Cell Line ◽  
Schwann Cells ◽  
Peripheral Nerve Regeneration ◽  
Cell Behavior ◽  
Cell Models ◽  
Migratory Activity ◽  
Pathological Conditions

The neuregulin1/ErbB system plays an important role in Schwann cell behavior both in normal and pathological conditions. Upon investigation of the expression of the neuregulin1/ErbB systemin vitro, we explored the possibility to manipulate the system in order to increase the migration of Schwann cells, that play a fundamental role in the peripheral nerve regeneration. Comparison of primary cells and stable cell lines shows that both primary olfactory bulb ensheathing cells and a corresponding cell line express ErbB1-ErbB2 and neuregulin1, and that both primary Schwann cells and a corresponding cell line express ErbB2-ErbB3, while only primary Schwann cells express neuregulin1. To interfere with the neuregulin1/ErbB system, the soluble extracellular domain of the neuregulin1 receptor ErbB4 (ecto-ErbB4) was expressedin vitroin the neuregulin1 expressing cell line, and an unexpected increase in cell motility was observed.In vitroexperiments suggest that the back signaling mediated by the transmembrane neuregulin1 plays a role in the migratory activity induced by ecto-ErbB4. These results indicate that ecto-ErbB4 could be usedin vivoas a tool to manipulate the neuregulin1/ErbB system.

scholarly journals The semantics of microglia activation: neuroinflammation, homeostasis, and stress

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Samuel C. Woodburn ◽  
Justin L. Bollinger ◽  
Eric S. Wohleb
Keyword(s):  
Microglia Activation ◽  
Immune Functions ◽  
Neuronal Function ◽  
Stress Effects ◽  
Pathological Conditions ◽  
Microglial Phenotype ◽  
And Function ◽  
And Behavior

AbstractMicroglia are emerging as critical regulators of neuronal function and behavior in nearly every area of neuroscience. Initial reports focused on classical immune functions of microglia in pathological contexts, however, immunological concepts from these studies have been applied to describe neuro-immune interactions in the absence of disease, injury, or infection. Indeed, terms such as ‘microglia activation’ or ‘neuroinflammation’ are used ubiquitously to describe changes in neuro-immune function in disparate contexts; particularly in stress research, where these terms prompt undue comparisons to pathological conditions. This creates a barrier for investigators new to neuro-immunology and ultimately hinders our understanding of stress effects on microglia. As more studies seek to understand the role of microglia in neurobiology and behavior, it is increasingly important to develop standard methods to study and define microglial phenotype and function. In this review, we summarize primary research on the role of microglia in pathological and physiological contexts. Further, we propose a framework to better describe changes in microglia1 phenotype and function in chronic stress. This approach will enable more precise characterization of microglia in different contexts, which should facilitate development of microglia-directed therapeutics in psychiatric and neurological disease.

Molecular identification and function characterization of four finTRIM genes from the immortal fish cell line, EPC

2020 ◽  
Vol 113 ◽  
pp. 103775
Author(s):  
Yi-Lin Li ◽  
Xiang Zhao ◽  
Xiu-Ying Gong ◽  
Cheng Dan ◽  
Jian-Fang Gui ◽  
...  
Keyword(s):  
Cell Line ◽  
Molecular Identification ◽  
Fish Cell Line ◽  
Fish Cell ◽  
Function Characterization ◽  
And Function

scholarly journals Mitochondrial Fusion Protein Mfn2 and Its Role in Heart Failure

2021 ◽  
Vol 8 ◽  
Author(s):  
Lei Chen ◽  
Bilin Liu ◽  
Yuan Qin ◽  
Anqi Li ◽  
Meng Gao ◽  
...  
Keyword(s):  
Heart Failure ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Mitochondrial Fusion ◽  
Mitochondrial Outer Membrane ◽  
Mitochondrial Quality Control ◽  
Mitofusin 2 ◽  
Multifunctional Protein ◽  
Pathological Conditions ◽  
And Function

Mitofusin 2 (Mfn2) is a transmembrane GTPase located on the mitochondrial outer membrane that contributes to mitochondrial network regulation. It is an essential multifunctional protein that participates in various biological processes under physical and pathological conditions, including mitochondrial fusion, reticulum–mitochondria contacts, mitochondrial quality control, and apoptosis. Mfn2 dysfunctions have been found to contribute to cardiovascular diseases, such as ischemia-reperfusion injury, heart failure, and dilated cardiomyopathy. Here, this review mainly focuses on what is known about the structure and function of Mfn2 and its crucial role in heart failure.

scholarly journals NMRK2 Gene Is Upregulated in Dilated Cardiomyopathy and Required for Cardiac Function and NAD Levels during Aging

2021 ◽  
Vol 22 (7) ◽  
pp. 3534
Author(s):  
Cynthia Tannous ◽  
Robin Deloux ◽  
Ahmed Karoui ◽  
Nathalie Mougenot ◽  
Dean Burkin ◽  
...  
Keyword(s):  
Dilated Cardiomyopathy ◽  
Therapeutic Potential ◽  
Pressure Overload ◽  
Concentric Hypertrophy ◽  
Cardiac Phenotype ◽  
Protein Isoform ◽  
Male Sex ◽  
And Function ◽  
Coenzyme Biosynthesis

Dilated cardiomyopathy (DCM) is a disease of multifactorial etiologies, the risk of which is increased by male sex and age. There are few therapeutic options for patients with DCM who would benefit from identification of common targetable pathways. We used bioinformatics to identify the Nmrk2 gene involved in nicotinamide adenine dinucleotde (NAD) coenzyme biosynthesis as activated in different mouse models and in hearts of human patients with DCM while the Nampt gene controlling a parallel pathway is repressed. A short NMRK2 protein isoform is also known as muscle integrin binding protein (MIBP) binding the α7β1 integrin complex. We investigated the cardiac phenotype of Nmrk2-KO mice to establish its role in cardiac remodeling and function. Young Nmrk2-KO mice developed an eccentric type of cardiac hypertrophy in response to pressure overload rather than the concentric hypertrophy observed in controls. Nmrk2-KO mice developed a progressive DCM-like phenotype with aging, associating eccentric remodeling of the left ventricle and a decline in ejection fraction and showed a reduction in myocardial NAD levels at 24 months. In agreement with involvement of NMRK2 in integrin signaling, we observed a defect in laminin deposition in the basal lamina of cardiomyocytes leading to increased fibrosis at middle age. The α7 integrin was repressed at both transcript and protein level at 24 months. Nmrk2 gene is required to preserve cardiac structure and function, and becomes an important component of the NAD biosynthetic pathways during aging. Molecular characterization of compounds modulating this pathway may have therapeutic potential.

scholarly journals Characterization of prostaglandin and thromboxane receptors expressed on a megakaryoblastic leukemia cell line, MEG-01s

Blood ◽  
1991 ◽  
Vol 78 (9) ◽  
pp. 2328-2336 ◽  
Author(s):  
T Watanabe ◽  
Y Yatomi ◽  
S Sunaga ◽  
I Miki ◽  
A Ishii ◽  
...  
Keyword(s):  
Cell Line ◽  
Binding Sites ◽  
Leukemia Cell ◽  
Leukemia Cell Line ◽  
Camp Production ◽  
Megakaryoblastic Leukemia ◽  
Ic50 Values ◽  
Thromboxane Receptors ◽  
And Function

MEG-01s, an established human megakaryoblastic leukemia cell line, exhibited specific high-affinity binding sites for [3H]iloprost, a stable prostaglandin (PG) I2 analogue, for [3H]SQ-29548, a stable thromboxane (TX) A2 antagonist and, for [3H]PGE2/PGE1, but not for [3H]PGD2. In the MEG-01s cells, iloprost/PGI2, or PGE1 stimulated cAMP production with ED50 values practically identical to the IC50 values for the [3H] iloprost binding. STA2 and U46619, TXA2/PGH2 agonists, PGE2/PGE1, iloprost/PGI2, and thrombin elevated the intracellular concentrations of Ca2+ ([Ca2+]i), as determined by Fura-2 fluorescence signals. Elevation of [Ca2+]i by PGE2/PGE1 and iloprost, but not that by TX-agonists or thrombin, was totally dependent on the presence of extracellular Ca2+. This effect by PGE2/PGE1 was partially inhibited by prior treatment of the cells with islet-activating protein (IAP), while that by TX-agonists or by PGI2/iloprost was not affected. We tentatively conclude from these results that: (1) MEG-01s cells express (a) PGI2/PGE1 receptor(s) coupled to adenylate cyclase and Ca2+ influx, a TXA2/PGH2 receptor coupled to the phosphatidylinositol-turnover-Ca2+ system, and the PGE2/PGE1 receptor coupled to Ca2+ influx; (2) the receptors for TXA2/PGH2 and iloprost and those for PGE2/PGE1 and thrombin are coupled to IAP-insensitive and IAP-sensitive GTP-binding proteins, respectively, and function in a different manner to elevate [Ca2+]i. Thus, the MEG-01s cell line is a pertinent model for studying eicosanoid receptor-mediated signal transduction in platelet/megakaryocyte systems.

scholarly journals Characterization of Mutational Status, Spheroid Formation, and Drug Response of a New Genomically-Stable Human Ovarian Clear Cell Carcinoma Cell Line, 105C

Cells ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 2408
Author(s):  
Bart Kolendowski ◽  
Yudith Ramos Valdes ◽  
Hal Hirte ◽  
Hiroaki Itamochi ◽  
Wonjae Lee ◽  
...  
Keyword(s):  
Suspension Culture ◽  
Cell Carcinoma ◽  
Cell Line ◽  
Clear Cell ◽  
Clear Cell Carcinoma ◽  
Gynecological Cancer ◽  
Model Systems ◽  
Ovarian Clear Cell Carcinoma ◽  
Mtor Inhibition

Ovarian clear cell carcinoma (OCCC) is a rare subtype of gynecological cancer for which well-characterized and authenticated model systems are scarce. We provide an extensive characterization of ‘105C’, a cell line generated from an adenocarcinoma of the clear cell histotype using targeted next-generation sequencing, cytogenetic microarrays, along with analyses of AKT/mTOR signaling. We report that that the 105C cell line is a bona fide OCCC cell line, carrying PIK3CA, PTEN, and ARID1A gene mutations, consistent with OCCC, yet maintain a stable genome as reflected by low copy number variation. Unlike KOC-7c, TOV-21G, and RMG-V OCCC lines also mutated for the above genes, the 105C cells do not carry mutations in mismatch repair genes. Importantly, we show that 105C cells exhibit greater resistance to mTOR inhibition and carboplatin treatment compared to 9 other OCCC cell lines in 3D spheroid cultures. This resistance may be attributed to 105C cells remaining dormant in suspension culture which surprisingly, contrasts with several other OCCC lines which continue to proliferate in long-term suspension culture. 105C cells survive xenotransplantation but do not proliferate and metastasize. Collectively, we show that the 105C OCCC cell line exhibits unique properties useful for the pre-clinical investigation of OCCC pathobiology.

scholarly journals Integrative and conjugative elements and their hosts: composition, distribution, and organization

2017 ◽  
Author(s):  
Jean Cury ◽  
Marie Touchon ◽  
Eduardo P. C. Rocha
Keyword(s):  
Large Scale ◽  
Functional Organization ◽  
Sequence Similarity ◽  
Phylogenetic Analyses ◽  
Model Systems ◽  
Integrative And Conjugative Elements ◽  
Scale Characterization ◽  
And Function ◽  
Sequence Similarity Networks

AbstractConjugation of single-stranded DNA drives horizontal gene transfer between bacteria and was widely studied in conjugative plasmids. The organization and function of integrative and conjugative elements (ICE), even if they are more abundant, was only studied in a few model systems. Comparative genomics of ICE has been precluded by the difficulty in finding and delimiting these elements. Here, we present the results of a method that circumvents these problems by requiring only the identification of the conjugation genes and the species’ pan-genome. We delimited 200 ICEs and this allowed the first large-scale characterization of these elements. We quantified the presence in ICEs of a wide set of functions associated with the biology of mobile genetic elements, including some that are typically associated with plasmids, such as partition and replication. Protein sequence similarity networks and phylogenetic analyses show that ICEs are modular and that their gene repertoires can be grouped in function of their conjugation types, even if integrases were shown to be paraphyletic relative to the latter. We show that there are general trends in the functional organization of genes within ICEs and of ICEs within the bacterial chromosome paving the way for future functional and evolutionary analyses.

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