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American Journal of Physiology. Cell Physiology

Katsuhito Matsuki, Daiki Kato, Masashi Takemoto, Yoshiaki Suzuki, Hisao Yamamura, Susumu Ohya, Hiroshi Takeshima, Yuji Imaizumi
Physiological functions of type 3 ryanodine receptors (RyR3) in smooth muscle (SM) tissues are not well understood, in spite of their wide expression. However, the short isoform of RyR3 is known to be a dominant negative variant (DN-RyR3), which may negatively regulate functions of both RyR2 and full length (FL)-RyR3 by forming hetero-tetramers. Here, functional roles of RyR3 in the regulation of Ca2+ signaling in mesenteric artery SM cells (MASMCs) were examined using RyR3 homozygous knockout mice (RyR3-/- )...
March 14, 2018: American Journal of Physiology. Cell Physiology
Minglin Pan, Ying Han, Aninda Basu, Anzhi Dai, Rui Si, Conor Willson, Angela Balistrieri, Brian T Scott, Ayako Makino
Coronary microvascular rarefaction due to endothelial cell (EC) dysfunction is one of the causes of increased morbidity and mortality in diabetes. Coronary ECs in diabetes are more apoptotic due partly to mitochondrial calcium overload. This study was designed to investigate the role of hexokinase 2 (HK2, an endogenous inhibitor of voltage-dependent anion channel) in coronary endothelial dysfunction in type 2 diabetes. We used mouse coronary ECs (MCECs) isolated from type 2 diabetic mice and human coronary ECs (HCECs) from type 2 diabetic patients to examine protein levels and mitochondrial functions...
March 7, 2018: American Journal of Physiology. Cell Physiology
Xiaojuan Xu, Mengmeng Guo, Na Zhang, Shoudong Ye
Although long noncoding RNAs (lncRNAs) are emerging as new modulators in the fate decision of pluripotent stem cells, the functions of specific lncRNAs remain unclear. Here, we found that telomeric RNA (TERRA or TelRNA), one type of lncRNAs, is highly expressed in mouse embryonic stem cells (mESCs), but declines significantly upon differentiation. TERRA is induced by the Wnt/β-catenin signaling pathway and can reproduce its self-renewal-promoting effect when overexpressed. Further studies revealed that TCF3 is a potential downstream target of TERRA and mediates the effect of TERRA in mESC maintenance...
March 7, 2018: American Journal of Physiology. Cell Physiology
Takayuki Uchida, Yoshihiro Sakashita, Kanako Kitahata, Yui Yamashita, Chisato Tomida, Yuki Kimori, Akio Komatsu, Katsuya Hirasaka, Ayako Ohno, Reiko Nakao, Atsushi Higashitani, Akira Higashibata, Noriaki Ishioka, Toru Shimazu, Takeshi Kobayashi, Yuushi Okumura, Inho Choi, Motoko Oarada, Edward M Mills, Shigetada Teshima-Kondo, Shin'ichi Takeda, Eiji Tanaka, Keiji Tanaka, Masahiro Sokabe, Takeshi Nikawa
Unloading-mediated muscle atrophy is associated with increased reactive oxygen species (ROS) production. We previously demonstrated that elevated ubiquitin ligase casitas B-lineage lymphoma-b (Cbl-b) resulted in the loss of muscle volume [Nakao R. et al. Mol Cell Biol, 29: 4798-4811, 2009]. However, the pathological role of ROS production associated with unloading-mediated muscle atrophy still remains unknown. Here, we showed the ROS-mediated signal transduction caused by microgravity or its simulation contributes to Cbl-b expression...
March 7, 2018: American Journal of Physiology. Cell Physiology
Ngonidzashe B Madungwe, Yansheng Feng, Mihaela Lie, Nathalie Tombo, Li Liu, Ferdinand Kaya, Jean C Bopassa
Mitofilin is an inner membrane protein that has been defined as a mitochondria-shaping protein in controlling and maintaining mitochondrial cristae structure and remodeling. We determined the role of mitofilin in cell survival by investigating the mechanism underlying mitofilin knockdown-induced cell death by apoptosis. Cultured H9c2 myoblasts and HEK 293 cells were treated with mitofilin siRNA or scrambled siRNA for 24 hours. Cell death (apoptosis), caspase 3 activity, and cell cycle phases were assessed by flow cytometry, while cytochrome C release and intracellular ATP production were measured by ELISA...
February 28, 2018: American Journal of Physiology. Cell Physiology
Hongyu Tan, Liang Zhao, Ruipeng Song, Yilin Liu, Limin Wang
Aberrant nucleus pulposus cell proliferation is implicated in the development of intervertebral disc degeneration (IDD). Recent studies have suggested that long non-coding RNAs (lncRNAs) can modulate cell proliferation in several pathological conditions. Here, we indicated that the expression of SNHG1 was upregulated in IDD tissues compared to the control tissues and higher SNHG1 expression was associated with disc degeneration grade. In addition, we showed that ectopic expression of SNHG1 promoted nucleus pulposus (NP) cell proliferation and increased the PCNA and cyclin D1 expression in NP cell...
February 21, 2018: American Journal of Physiology. Cell Physiology
Ming Lu, Jian-Ri Li, Lery Alvarez-Lugo, Yan Li, Shan Yu, Xuanhao Li, Benkang Shi, Toby C Chai
Bladder urothelium plays an active role in response to bacterial infection. There is little known about the electrophysiological activity in urothelial cells in this process. We used a non-enzymatic method to isolate bladder urothelial tissue and to patch clamp umbrella cells in situ. A 200pS conductance potassium (K+ ) channel was detected from female C57BL6 mice. Of 58 total patches, 17.2% patches displayed the 200pS K+ conductance channel. This K+ conductance channel showed Ca2+ sensitivity and voltage dependence...
February 21, 2018: American Journal of Physiology. Cell Physiology
Yun-Xiao Zhou, Chuan Wang, Li-Wei Mao, Yan-Li Wang, Li-Qun Xia, Wei Zhao, Jie Shen, Jun Chen
Background LncRNA homeobox (HOX) transcript antisense intergenic RNA (HOTAIR) has been confirmed to involve in the tumorigenic progression of endometrial carcinoma (EC). However, the molecular mechanism of HOTAIR in EC are not fully elucidated. Methods The expression of HOTAIR and miR-646 in human EC tissues was determined by qRT-PCR. The effect of miR-646 on EC cells was assessed by the cell viability, migration and invasion using CCK-8 assays and transwell assays. RIP assays and RNA pull-down assays were performed to explore the interaction between HOTAIR and miR-646...
February 21, 2018: American Journal of Physiology. Cell Physiology
Cristian A Pocognoni, Ekaterina G Viktorova, John Wright, Justyna M Meissner, Garrett Sager, Eunjoo Lee, George A Belov, Elizabeth Sztul
Cellular life requires the activation of the ADP-ribosylation factors (ARFs) by GBF1, a guanine nucleotide exchange factor (GEF) with a highly conserved catalytic Sec7 domain (Sec7d). In addition to the Sec7d, GBF1 contains other conserved domains whose functions remain unclear. Here, we focus on HDS2 (homology downstream of the Sec7d 2) domain because the L1246R substitution within the HDS2 α-helix 5 of the zebrafish GBF1 ortholog causes vascular hemorrhaging and embryonic lethality (13). To dissect the structure/function relationships within HDS2, we generated six variants, in which the most conserved residues within α-helices 1, 2, 4 and 6 were mutated to alanines...
February 14, 2018: American Journal of Physiology. Cell Physiology
Itamar Couto Guedes de Jesus, Sergio Scalzo, Fabiana Alves, Kariny Marques, Cibele Rocha-Resende, Michael Bader, Robson A Souza Santos, Silvia Guatimosim
The renin-angiotensin system (RAS) plays a pivotal role in the pathogenesis of cardiovascular diseases. New members of this system have been characterized and shown to have biologically relevant actions. Alamandine and its receptor MrgD are recently identified components of RAS. In the cardiovascular system alamandine actions included vasodilation, antihypertensive and anti-fibrosis effects. Currently, the actions of alamandine on cardiomyocytes are unknown. Here our goal was twofold: (1) to unravel the signaling molecules activated by the alamandine/MrgD axis in cardiomyocytes; (2) to evaluate the ability of this axis to prevent against Angiotensin II (Ang II)-induced hypertrophy...
February 14, 2018: American Journal of Physiology. Cell Physiology
Özgün Babur, Anh T P Ngo, Rachel A Rigg, Jiaqing Pang, Zhoe T Rub, Ariana E Buchanan, Annachiara Mitrugno, Larry L David, Owen J T McCarty, Emek Demir, Joseph E Aslan
Upon encountering physiological cues associated with damaged or inflamed endothelium, blood platelets set forth intracellular responses to ultimately support hemostatic plug formation and vascular repair. To gain insights into the molecular events underlying platelet function, we used a combination of interactome, pathway analysis and other systems biology tools to analyze associations amongst proteins functionally modified by reversible phosphorylation upon platelet activation. While an interaction analysis mapped out a relative organization of intracellular mediators in platelet signaling, pathway analysis revealed directional signaling relations around protein kinase C (PKC) isoforms and mitogen activated protein kinases (MAPKs) associated with platelet cytoskeletal dynamics, inflammatory responses and hemostatic function...
February 7, 2018: American Journal of Physiology. Cell Physiology
Leonit Kiriaev, Sindy Kueh, John W Morley, Kathryn N North, Peter J Houweling, Stewart I Head
A striking pathological feature of dystrophinopathies is the presence of morphologically abnormal branched skeletal muscle fibers. The deterioration of muscle contractile function in Duchenne muscular dystrophy is accompanied by both an increase in number and complexity of these branched fibers. We propose that when number and complexity of branched fibers reaches a critical threshold, "tipping point" the branches in and of themselves are the site of contraction-induced rupture. In the present study, we use the dystrophic mdx mouse and littermate controls to study the pre-diseased dystrophic fast-twitch EDL muscle at 2-3-weeks, the peak myonecrotic phase at 6-9 weeks and finally "old" at 58-112 weeks...
February 7, 2018: American Journal of Physiology. Cell Physiology
Mark D Parker
The SLC4 family Cl- -HCO3 - exchangers (AE1, AE2, and AE3) and Na+ /HCO3 - cotransporters (NBCe1, NBCe2, NBCn1, and NBCn2) contribute to a variety of vital physiological processes including pH regulation and epithelial fluid secretion. Accordingly, their dysfunction can have devastating effects. Disorders such as epilepsy, hemolytic anemia, glaucoma, hearing loss, osteopetrosis and renal tubular acidosis are all genetically linked to SLC4-family gene loci. This review summarizes how studies of Slc4-modified mice have enhanced our understanding of the etiology of SLC4-linked pathologies and the interpretation of genetic linkage studies...
January 31, 2018: American Journal of Physiology. Cell Physiology
Helene H Jensen, Mikkel R Holst, Frédéric H Login, Jeanette J Morgen, Lene N Nejsum
Aquaporin-5 (AQP5) is a plasma membrane water channel mainly expressed in secretory glands. Increased expression of AQP5 is observed in multiple cancers, including breast cancer, where high expression correlates with the degree of metastasis and poor prognosis. Moreover, studies in cancer cells have suggested that AQP5 activates Ras signaling, drives morphological changes, and in particular increased invasiveness. To design intervention strategies, it is of utmost importance to characterize and dissect the cell biological changes induced by altered AQP5 expression...
January 31, 2018: American Journal of Physiology. Cell Physiology
Puneet Raman, Lakshmi Madhavpeddi, Rayna J Gonzales
Vascular basal cyclooxygenase-2 (COX-2) expression and activity can be induced by endotoxin, hypoxia, or ischemia. During vascular pathologies such as atherosclerosis, increases in COX-2 activity result in prostanoid production, a contributor to the development and progression of vascular inflammation leading to unstable atherosclerotic plaques and increased risk for thrombotic events. Recent studies demonstrate that select free fatty acids, such as palmitate, can act as pro-inflammatory mediators. However, the effect of palmitate on COX-2 expression and activity, and its impact on the development and progression of vascular inflammation is not well elucidated...
January 31, 2018: American Journal of Physiology. Cell Physiology
Benard Ogola, Yong Zhang, Laxmi Iyer, Thomas Thekkumkara
Studies have demonstrated the therapeutic potential of estrogen metabolite 2-Methoxyestradiol (2ME2) in several cardiovascular disorders, including hypertension. However, the exact mechanism(s) remains unknown. In this study, primary rat aortic smooth muscle cells (RASMCs) were exposed to 2ME2 and angiotensin type 1 receptor (AT1R) expression, function and associated signaling pathways were evaluated. 2ME2 downregulated AT1R expression in a concentration and time-dependent manner, which was correlated with reduced mRNA expression...
January 24, 2018: American Journal of Physiology. Cell Physiology
Yotesawee Srisomboon, Nathan A Zaidman, Peter J Maniak, Chatsri Deachapunya, Scott M O'Grady
The objective of this study was to determine the molecular identity of ion channels involved in K+ secretion by the mammary epithelium and to examine their regulation by purinoceptor agonists. Apical membrane voltage clamp experiments were performed on human mammary epithelial cells where the basolateral membrane was exposed to the pore-forming antibiotic amphotericin B dissolved in a solution with intracellular-like ionic composition. Addition of the Na+ channel inhibitor benzamil reduced the basal current, consistent with inhibition of Na+ uptake across the apical membrane whereas the KCa 3...
January 24, 2018: American Journal of Physiology. Cell Physiology
Muriel Nobles, David Montaigne, Sonia Sebastian, Lutz Birnbaumer, Andrew Tinker
G-protein gated inwardly rectifying K+ (GIRK) channels are the major inwardly rectifying K+ currents in cardiac atrial myocytes and an important determinant of atrial electrophysiology. Inhibitory G-protein alpha subunits can both mediate activation via acetylcholine but can also supress basal currents in the absence of agonist. We studied this phenomenon using whole cell patch clamping in murine atria from mice with global genetic deletion of Gαi2, combined deletion of Gαi1/Gαi3 and littermate controls...
January 17, 2018: American Journal of Physiology. Cell Physiology
Inga Baasch Christensen, Esben Nees Mogensen, Helle Hasager Damkier, Jeppe Praetorius
The choroid plexus epithelial cells (CPECs) belong to a small group of polarized cells, where the Na+,K+-ATPase is expressed in the luminal membrane. The basic polarity of the cells is, therefore, still debated. We investigated the subcellular distribution of an array of proteins known to play fundamental roles in either in establishing and maintaining basic cell polarity or the polarized delivery and recycling of plasma membrane proteins. Immunofluorescence histochemical analysis was applied to determine the subcellular localization of apical and basolateral membrane determinants...
January 10, 2018: American Journal of Physiology. Cell Physiology
Mitra Farnoodian, Christine M Sorenson, Nader Sheibani
Age related macular degeneration (AMD) is the leading cause of vision loss among the elderly population, and is associated with severe macular degeneration and choroidal neovascularization (CNV). Although the pathogenesis of AMD is associated with choroidal dysfunction and CNV, the detailed underlying mechanisms remain unresolved. Altered production of pigment epithelium derived factor (PEDF), a neuroprotective and anti-angiogenic factor, contributes to CNV. Furthermore, exogenous PEDF mitigates angiogenesis in preclinical CNV models...
January 10, 2018: American Journal of Physiology. Cell Physiology
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