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https://www.readbyqxmd.com/read/29768048/hypoxic-stress-up-regulates-kir2-1-expression-by-a-pathway-including-hypoxic-inducible-factor-1-and-dynamin2-in-brain-capillary-endothelial-cells
#1
Hideto Yamamura, Yoshiaki Suzuki, Hisao Yamamura, Kiyofumi Asai, Wayne Giles, Yuji Imaizumi
Brain capillary endothelial cells (BCECs) play a central role in maintenance of blood-brain barrier (BBB) function and therefore are essential for central nervous system homeostasis and integrity. Although brain ischemia damages BCECs and causes disruption of BBB, the related influence of hypoxia on BCECs is not well understood. Hypoxic stress can up-regulate functional expression of specific K+ currents in endothelial cells, e.g., Kir2.1 channels without any alterations in the mRNA level, in t-BBEC117, a cell line derived from bovine BCECs...
May 16, 2018: American Journal of Physiology. Cell Physiology
https://www.readbyqxmd.com/read/29719170/vegf-a-selectively-inhibits-flt1-ectodomain-shedding-independent-of-receptor-activation-and-receptor-endocytosis
#2
Nandita S Raikwar, Masabumi Shibuya, Christie P Thomas
Ectodomain shedding and regulated intracellular proteolysis can determine the fate or function of cell surface proteins. Flt1 or VEGFR1 is a high affinity cell surface VEGF-A receptor tyrosine kinase which is constitutively cleaved to release an N-terminal VEGF-A binding ectodomain which once shed can antagonize the effects of VEGF-A in the extracellular milieu. We evaluated the effect of VEGF-A on FLT1 cleavage in native cells and in transient and stable expression systems. We demonstrate that VEGF-A inhibits FLT1 ectodomain cleavage in a time- and dose-dependent manner while VEGF-A knockdown in HEK293 cells increases ectodomain shedding...
May 2, 2018: American Journal of Physiology. Cell Physiology
https://www.readbyqxmd.com/read/29614089/hippocampal-protein-kinase-d1-is-necessary-for-dhpg-induced-learning-and-memory-impairments-in-rats
#3
Wei Wang, Florian Duclot, Bradley R Groveman, Nicole Carrier, Haifa Qiao, Xiao-Qian Fang, Hui Wang, Wenkuan Xin, Xing-Hong Jiang, Michael W Salter, Xin-Sheng Ding, Mohamed Kabbaj, Xian-Min Yu
BACKGROUND: Understanding molecular mechanisms underlying the induction of learning and memory impairments remains a challenge. Recent investigations have shown that the activation of group I mGluRs (mGluR1 and mGluR5) in cultured hippocampal neurons by application of (S)-3,5-Dihydroxyphenylglycine (DHPG) causes the regulated internalization of N-methyl-D-aspartate receptors (NMDARs), which subsequently activates protein kinase D1 (PKD1). Through phosphorylating the C-terminals of the NMDAR GluN2 subunits, PKD1 down-regulates the activity of remaining (non-internalized) surface NMDARs...
2018: PloS One
https://www.readbyqxmd.com/read/29569954/nitrosative-stress-uncovers-potent-%C3%AE-2-adrenergic-receptor-linked-vasodilation-further-enhanced-by-blocking-clathrin-endosome-formation
#4
Mary D Frame, Anthony M Dewar, Rhodora C Calizo, Adroniqui Qifti, Suzanne F Scarlata
This study investigated the effect of nitroprusside (SNP) pre-exposure on vasodilation via the beta-adrenergic receptor system. SNP was used as a nitrosative / oxidative pro-inflammatory insult. Small arterioles were visualized by intravital microscopy in the hamster cheek pouch tissue (isoflurane, N=45). Control dilation to isoproterenol (EC50 10-7 mol/L) became bi-phasic as a function of concentration following 2 min exposure to SNP (10-4 M), with a pico-molar dilation uncovered, and the micro-molar dilation attenuated...
March 23, 2018: American Journal of Physiology. Heart and Circulatory Physiology
https://www.readbyqxmd.com/read/29467541/inhibition-of-endo-lysosomal-function-exacerbates-vascular-calcification
#5
Yujun Cai, Xue-Lin Wang, Alyssa M Flores, Tonghui Lin, Raul J Guzman
Vascular calcification is a pathologic response to mineral imbalances and is prevalent in atherosclerosis, diabetes mellitus, and chronic kidney disease. When located in the media, it is highly associated with increased cardiovascular morbidity and mortality, particularly in patients on dialysis. Vascular calcification is tightly regulated and controlled by a series of endogenous factors. In the present study, we assess the effects of lysosomal and endosomal inhibition on calcification in vascular smooth muscle cells (VSMCs) and aortic rings...
February 21, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29321326/the-amino-terminus-of-herpes-simplex-virus-1-glycoprotein-k-gk-is-required-for-gb-binding-to-akt-release-of-intracellular-calcium-and-fusion-of-the-viral-envelope-with-plasma-membranes
#6
Farhana Musarrat, Nithya Jambunathan, Paul J F Rider, V N Chouljenko, K G Kousoulas
Previously, we have shown that the amino terminus of glycoprotein K (gK) binds to the amino terminus of gB and that deletion of the amino-terminal 38 amino acids of gK prevents herpes simplex virus 1 (HSV-1) infection of mouse trigeminal ganglia after ocular infection and virus entry into neuronal axons. Recently, it has been shown that gB binds to Akt during virus entry and induces Akt phosphorylation and intracellular calcium release. Proximity ligation and two-way immunoprecipitation assays using monoclonal antibodies against gB and Akt-1 phosphorylated at S473 [Akt-1(S473)] confirmed that HSV-1(McKrae) gB interacted with Akt-1(S473) during virus entry into human neuroblastoma (SK-N-SH) cells and induced the release of intracellular calcium...
March 15, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29320741/endocytosis-of-k-atp-channels-drives-glucose-stimulated-excitation-of-pancreatic-%C3%AE-cells
#7
Young-Eun Han, Jung Nyeo Chun, Min Jeong Kwon, Young-Sun Ji, Myong-Ho Jeong, Hye-Hyun Kim, Sun-Hyun Park, Jong Cheol Rah, Jong-Sun Kang, Suk-Ho Lee, Won-Kyung Ho
Insulin secretion from pancreatic β cells in response to high glucose (HG) critically depends on the inhibition of KATP channel activity in HG. It is generally believed that HG-induced effects are mediated by the increase in intracellular ATP, but here, we showed that, in INS-1 cells, endocytosis of KATP channel plays a major role. Upon HG stimulation, resting membrane potential depolarized by 30.6 mV (from -69.2 to -38.6 mV) and KATP conductance decreased by 91% (from 0.243 to 0.022 nS/pF), whereas intracellular ATP was increased by only 47%...
January 9, 2018: Cell Reports
https://www.readbyqxmd.com/read/29298696/enhanced-human-enterovirus-71-infection-by-endocytosis-inhibitors-reveals-multiple-entry-pathways-by-enterovirus-causing-hand-foot-and-mouth-diseases
#8
Meichun Yuan, Jingjing Yan, Jingna Xun, Chong Chen, Yuling Zhang, Min Wang, Wenqi Chu, Zhigang Song, Yunwen Hu, Shuye Zhang, Xiaoyan Zhang
BACKGROUND: Human enterovirus 71 (EV71) was previously known to enter cells through clathrin or caveolar mediated endocytic pathways. However, we observed chlorpromazine (CPZ) or dynasore (DNS), which inhibit clathrin and dynamin mediated endocytosis, did not suppress EV71 cell entry in particular cell types. So the current knowledge of entry mechanisms by EV71 is not complete. METHODS: Viral infection was examined by flow cytometry or end-point dilution assays...
January 3, 2018: Virology Journal
https://www.readbyqxmd.com/read/29296945/the-class-i-scavenger-receptor-cd163-promotes-internalization-of-adamts13-by-macrophages
#9
Fabian C Verbij, Nicoletta Sorvillo, Paul H P Kaijen, Johana Hrdinova, Ivan Peyron, Rob Fijnheer, Anja Ten Brinke, Alexander B Meijer, Floris P J van Alphen, Timo K van den Berg, Jonas J H Graversen, Soren K Moestrup, Jan Voorberg
Internalization of ADAMTS13 by macrophages may contribute to its clearance from the circulation. Here we investigated endocytic mechanisms that contribute to the uptake of ADAMTS13 by macrophages. Human monocyte-derived macrophages were used to monitor the uptake of fluorescently labeled recombinant ADAMTS13 by flow cytometry. Internalization of ADAMTS13 was blocked upon addition of the cell-permeable dynamin inhibitor dynasore. Partial blocking of ADAMTS13 uptake was observed by using mannan; however, uptake was not affected by an antibody that blocked binding to the macrophage mannose receptor CD206, which suggests that other endocytic receptors contribute to the internalization of ADAMTS13 by macrophages...
January 24, 2017: Blood Advances
https://www.readbyqxmd.com/read/29179200/stress-kinase-regulation-of-task-1-and-task-3
#10
Susanne Rinné, Aytug K Kiper, Constanze Schmidt, Beatriz Ortiz-Bonnin, Simone Zwiener, Guiscard Seebohm, Niels Decher
BACKGROUND/AIMS: TASK channels belong to the two-pore-domain potassium (K2P) channel family. TASK-1 is discussed to contribute to chronic atrial fibrillation (AFib) and has been together with uncoupling protein 1 found as a marker protein of brown adipose tissue (BAT) fat. In addition, TASK-1 was linked in a genome-wide association study to an increased body mass index. A recent study showed that TASK-1 inhibition is causing obesity in mice by a BAT whitening and that these effects are linked to the mineralocorticoid receptor pathway, albeit the mechanism remained elusive...
2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29175207/dynasore-suppresses-proliferation-and-induces-apoptosis-of-the-non-small-cell-lung-cancer-cell-line-a549
#11
Feifei Shen, Junda Gai, Jilin Xing, Jingqian Guan, Lin Fu, Qingchang Li
Lung cancer is the leading cause of cancer death worldwide, and most of all cases are non-small-cell lung cancer. Lung cancer is associated with dysregulation of mitochondrial fusion and fission, and inhibition of the fission regulator Dynamin-related protein 1 (Drp1) reduces proliferation and increases apoptosis of lung cancer cells. Dynasore is a small molecule non-selective inhibitor of the GTPase activity of dynamin 1, dynamin 2, and Drp1 in vivo and in vitro. Here, we investigated the effects of dynasore on the proliferation and apoptosis of A549 lung cancer cells, alone and in combination with the chemotherapeutic drug cisplatin...
January 1, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29152606/ethanol-induced-steatosis-involves-impairment-of-lipophagy-associated-with-reduced-dynamin2-activity
#12
Karuna Rasineni, Terrence M Donohue, Paul G Thomes, Li Yang, Dean J Tuma, Mark A McNiven, Carol A Casey
Background: Lipid droplets (LDs), the organelles central to alcoholic steatosis, are broken down by lipophagy, a specialized form of autophagy. Here, we hypothesize that ethanol administration retards lipophagy by down-regulating Dynamin 2 (Dyn2), a protein that facilitates lysosome re-formation, contributing to hepatocellular steatosis. Methods: Primary hepatocytes were isolated from male Wistar rats fed Lieber-DeCarli control or EtOH liquid diets for 6-8 wk. Hepatocytes were incubated in complete medium (fed) or nutrient-free medium (fasting) with or without the Dyn2 inhibitor Dynasore or the Src inhibitor SU6656...
August 2017: Hepatology Communications
https://www.readbyqxmd.com/read/29150487/dynamin-inhibitors-block-activation-of-mtorc1-by-amino-acids-independently-of-dynamin
#13
Avinash Persaud, Yann Cormerais, Jacques Pouyssegur, Daniela Rotin
mTORC1 plays a crucial role in protein synthesis and cell proliferation and growth. It is activated by growth factors and amino acids, including essential amino acids (EAAs), such as leucine; Leu enters cells via the Leu transporter LAT1-4F2hc (also known as SLC7A5-SLC3A2) and potentially via endocytosis. Here, we investigated the contribution of the different routes of Leu entry into cells to mTORC1 activation using pharmacological inhibitors and cells that lack LAT1 or dynamin-1, -2 and -3. Our results show that LAT1 is the major route of Leu entry into cells and mTORC1 activation (∼70%), whereas dynamin-dependent endocytosis and macropinocytosis contribute minimally to both (5-15%)...
January 4, 2018: Journal of Cell Science
https://www.readbyqxmd.com/read/29077063/p2x4-receptor-dependent-ca-2-influx-in-model-human-monocytes-and-macrophages
#14
Janice A Layhadi, Samuel J Fountain
Monocytes and macrophages express a repertoire of cell surface P2 receptors for adenosine 5'-triphosphate (ATP) a damage-associated molecular pattern molecule (DAMP), which are capable of raising cytoplasmic calcium when activated. This is achieved either through direct permeation (ionotropic P2X receptors) or by mobilizing intracellular calcium stores (metabotropic P2Y receptors). Here, a side-by-side comparison to investigate the contribution of P2X4 receptor activation in ATP-evoked calcium responses in model human monocytes and macrophages was performed...
October 27, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28957392/a-highly-sensitive-high-throughput-assay-for-dynamin-s-basal-gtpase-activity
#15
Aparna Mohanakrishnan, Triet Vincent M Tran, Meera Kumar, Hong Chen, Bruce A Posner, Sandra L Schmid
Clathrin-mediated endocytosis is the major pathway by which cells internalize materials from the external environment. Dynamin, a large multidomain GTPase, is a key regulator of clathrin-mediated endocytosis. It assembles at the necks of invaginated clathrin-coated pits and, through GTP hydrolysis, catalyzes scission and release of clathrin-coated vesicles from the plasma membrane. Several small molecule inhibitors of dynamin's GTPase activity, such as Dynasore and Dyngo-4a, are currently available, although their specificity has been brought into question...
2017: PloS One
https://www.readbyqxmd.com/read/28944482/molecular-mechanism-for-muscarinic-m1-receptor-mediated-endocytosis-of-twik-related-acid-sensitive-k-1-channels-in-rat-adrenal-medullary-cells
#16
Hidetada Matsuoka, Masumi Inoue
KEY POINTS: The muscarinic acetylcholine receptor (mAChR)-mediated increase in excitability in rat adrenal medullary cells is at least in part due to inhibition of TWIK (tandem of P domains in a weak inwardly rectifying K(+) channel)-related acid-sensitive K(+) (TASK)1 channels. In this study we focused on the molecular mechanism of mAChR-mediated inhibition of TASK1 channels. Exposure to muscarine resulted in a clathrin-dependent endocytosis of TASK1 channels following activation of the muscarinic M1 receptor (M1 R)...
November 15, 2017: Journal of Physiology
https://www.readbyqxmd.com/read/28880029/fluorescence-labelling-of-extracellular-vesicles-using-a-novel-thiol-based-strategy-for-quantitative-analysis-of-cellular-delivery-and-intracellular-traffic
#17
H D Roberts-Dalton, A Cocks, J M Falcon-Perez, E J Sayers, J P Webber, P Watson, A Clayton, A T Jones
Extracellular vesicles, including exosomes, are naturally derived nanovesicles generated in and released by numerous cell types. As extracellular entities they have the capacity to interact with neighbouring cells and distant tissues and affect physiological processes as well as being implicated in numerous diseases including tumorigenesis and neurodegeneration. They are also under intense investigation as delivery vectors for biotherapeutics. The ways in which EVs interact with recipient cells to influence cell physiology and deliver a macromolecular payload are at the early stages of exploration...
September 21, 2017: Nanoscale
https://www.readbyqxmd.com/read/28750371/mitochondrial-fission-inhibitors-suppress-endothelin-1-induced-artery-constriction
#18
Chang Chen, Jin-Lai Gao, Ming-Yu Liu, Shan-Liang Li, Xiu-Chen Xuan, Xin-Zi Zhang, Xi-Yue Zhang, Yuan-Yuan Wei, Chang-Lin Zhen, Jing Jin, Xin Shen, De-Li Dong
BACKGROUND/AIMS: Endothelin-1 is implicated in the pathogenesis of hypertension, but the underlying mechanisms remained elusive. Our previous study found that inhibition of mitochondrial fission of smooth muscle cells suppressed phenylephrine- and high K+-induced artery constriction. Here, we studied the effects of mitochondrial fission inhibitors on endothelin-1-induced vasoconstriction. METHODS: The tension of rat mesenteric arteries and thoracic aorta was measured by using a multi-wire myograph system...
2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28716920/vamp3-and-snap23-mediate-the-disturbed-flow-induced-endothelial-microrna-secretion-and-smooth-muscle-hyperplasia
#19
Juan-Juan Zhu, Yue-Feng Liu, Yun-Peng Zhang, Chuan-Rong Zhao, Wei-Juan Yao, Yi-Shuan Li, Kuei-Chun Wang, Tse-Shun Huang, Wei Pang, Xi-Fu Wang, Xian Wang, Shu Chien, Jing Zhou
Vascular endothelial cells (ECs) at arterial branches and curvatures experience disturbed blood flow and induce a quiescent-to-activated phenotypic transition of the adjacent smooth muscle cells (SMCs) and a subsequent smooth muscle hyperplasia. However, the mechanism underlying the flow pattern-specific initiation of EC-to-SMC signaling remains elusive. Our previous study demonstrated that endothelial microRNA-126-3p (miR-126-3p) acts as a key intercellular molecule to increase turnover of the recipient SMCs, and that its release is reduced by atheroprotective laminar shear (12 dynes/cm2 ) to ECs...
August 1, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28592413/recruitment-of-endosomal-signaling-mediates-the-forskolin-modulation-of-guinea-pig-cardiac-neuron-excitability
#20
Jean C Hardwick, Todd A Clason, John D Tompkins, Beatrice M Girard, Caitlin N Baran, Laura A Merriam, Victor May, Rodney L Parsons
Forskolin, a selective activator of adenylyl cyclase (AC), commonly is used to establish actions of G protein-coupled receptors (GPCRs) that are initiated primarily through activation of AC/cAMP signaling pathways. In the present study, forskolin was used to evaluate the potential role of AC/cAMP, which is a major signaling mechanism for the pituitary adenylate cyclase-activating polypeptide (PACAP)-selective PAC1 receptor, in the regulation of guinea pig cardiac neuronal excitability. Forskolin (5-10 µM) increases excitability in ~60% of the cardiac neurons...
August 1, 2017: American Journal of Physiology. Cell Physiology
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