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Myosin IIB

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https://www.readbyqxmd.com/read/28814507/systematic-gene-tagging-using-crispr-cas9-in-human-stem-cells-to-illuminate-cell-organization
#1
Brock Roberts, Amanda Haupt, Andrew Tucker, Tanya Grancharova, Joy Arakaki, Margaret A Fuqua, Angelique Nelson, Caroline Hookway, Susan A Ludmann, Irina A Mueller, Ruian Yang, Alan R Horwitz, Susanne M Rafelski, Ruwanthi N Gunawardane
We present a CRISPR/Cas9 genome editing strategy to systematically tag endogenous proteins with fluorescent tags in human induced pluripotent stem cells. To date we have generated multiple human iPSC lines with monoallelic GFP tags labeling 10 proteins representing major cellular structures. The tagged proteins include alpha tubulin, beta actin, desmoplakin, fibrillarin, nuclear lamin B1, non-muscle myosin heavy chain IIB, paxillin, Sec61 beta, tight junction protein ZO1, and Tom20. Our genome editing methodology using Cas9/crRNA ribonuclear protein and donor plasmid co-electroporation, followed by fluorescence-based enrichment of edited cells, typically resulted in <0...
August 16, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28769032/vestigial-like-2-contributes-to-normal-muscle-fiber-type-distribution-in-mice
#2
Masahiko Honda, Kyoko Hidaka, So-Ichiro Fukada, Ryo Sugawa, Manabu Shirai, Masahito Ikawa, Takayuki Morisaki
Skeletal muscle is composed of heterogeneous populations of myofibers that are classified as slow- and fast-twitch fibers. The muscle fiber-type is regulated in a coordinated fashion by multiple genes, including transcriptional factors and microRNAs (miRNAs). However, players involved in this regulation are not fully elucidated. One of the members of the Vestigial-like factors, Vgll2, is thought to play a pivotal role in TEA domain (TEAD) transcription factor-mediated muscle-specific gene expression because of its restricted expression in skeletal muscles of adult mice...
August 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28752107/muscle-histology-characterization-using-h-e-staining-and-muscle-fiber-type-classification-using-immunofluorescence-staining
#3
Chao Wang, Feng Yue, Shihuan Kuang
Muscle function is determined by its structure and fiber type composition. Here we describe a protocol to examine muscle histology and myofiber types using hematoxylin and eosin (H&E) and immunofluorescence staining, respectively. H&E stain nucleus in blue and cytoplasm in red, therefore allowing for morphological analyses, such as myofiber diameter, the presence of degenerated and regenerated myofibers, and adipocytes and fibrotic cells. Muscle fibers in adult skeletal muscles of rodents are classified into 4 subtypes based on the expression of myosin heavy chain proteins: Myh7 (type I fiber), Myh2 (type IIA fiber), Myh1 (type IIX fiber), Myh4 (type IIB fiber)...
May 20, 2017: Bio-protocol
https://www.readbyqxmd.com/read/28729432/tropomodulin1-controls-erythroblast-enucleation-via-regulation-of-f-actin-in-the-enucleosome
#4
Roberta B Nowak, Julien Papoin, David S Gokhin, Carla Casu, Stefano Rivella, Jeffrey M Lipton, Lionel Blanc, Velia M Fowler
Biogenesis of mammalian red blood cells requires nuclear expulsion by orthochromatic erythoblasts late in terminal differentiation (enucleation), but the mechanism is largely unexplained. Here, we employed high resolution confocal microscopy to analyze nuclear morphology and F-actin rearrangements during the initiation, progression and completion of mouse and human erythroblast enucleation in vivo Mouse erythroblast nuclei acquire a dumbbell-shaped morphology during enucleation, while human bone marrow erythroblast nuclei unexpectedly retain their spherical morphology...
July 20, 2017: Blood
https://www.readbyqxmd.com/read/28728382/effects-of-a-traditional-chinese-medicine-formula-and-its-extraction-on-muscle-fiber-characteristics-in-finishing-pigs-porcine-cell-proliferation-and-isoforms-of-myosin-heavy-chain-gene-expression-in-myocytes
#5
Qin Ping Yu, Ding Yuan Feng, Xiao Jun He, Fan Wu, Min Hao Xia, Tao Dong, Yi Hua Liu, Hui Ze Tan, Shi Geng Zou, Tao Zheng, Xian Hua Ou, Jian Jun Zou
Objective: This study evaluated the effects of a traditional Chinese medicine formula (TCMF) on muscle fiber characteristics in finishing pigs and the effects of the formula's extract (distilled water, ethyl acetate and petroleum ether extraction) on porcine cell proliferation and isoforms of MyHC gene expression in myocytes. Methods: XThe psoas major muscle was obtained from pigs at the end of the experiment. Muscle fiber characteristics in the psoas major muscle were analyzed using myosin ATPase staining...
June 26, 2017: Asian-Australasian Journal of Animal Sciences
https://www.readbyqxmd.com/read/28701425/self-sorting-of-nonmuscle-myosins-iia-and-iib-polarizes-the-cytoskeleton-and-modulates-cell-motility
#6
Maria S Shutova, Sreeja B Asokan, Shefali Talwar, Richard K Assoian, James E Bear, Tatyana M Svitkina
Nonmuscle myosin II (NMII) is uniquely responsible for cell contractility and thus defines multiple aspects of cell behavior. To generate contraction, NMII molecules polymerize into bipolar minifilaments. Different NMII paralogs are often coexpressed in cells and can copolymerize, suggesting that they may cooperate to facilitate cell motility. However, whether such cooperation exists and how it may work remain unknown. We show that copolymerization of NMIIA and NMIIB followed by their differential turnover leads to self-sorting of NMIIA and NMIIB along the front-rear axis, thus producing a polarized actin-NMII cytoskeleton...
July 12, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28642260/cigarette-smoke-disrupts-monolayer-integrity-by-altering-epithelial-cell-cell-adhesion-and-cortical-tension
#7
Kristine Nishida, Kieran A Brune, Nirupama Putcha, Pooja Mandke, Wanda K O'Neal, Danny Shade, Vasudha Srivastava, Menghen Wang, Hong Lam, Steven S An, M Bradley Drummond, Nadia N Hansel, Douglas N Robinson, Venkataramana K Sidhaye
Chronic obstructive pulmonary disease (COPD) is a major cause of morbidity and mortality. Cigarette smoke (CS) drives disease development and progression. The epithelial barrier is damaged by CS with increased monolayer permeability. However, the molecular changes that cause this barrier disruption and the interaction between adhesion proteins and the cytoskeleton are not well defined. We hypothesized that CS alters monolayer integrity by increasing cell contractility and decreasing cell adhesion in epithelia...
June 22, 2017: American Journal of Physiology. Lung Cellular and Molecular Physiology
https://www.readbyqxmd.com/read/28622884/the-effect-of-caloric-restriction-on-the-forelimb-skeletal-muscle-fibers-of-the-hypertrophic-myostatin-null-mice
#8
Mohamed I Elashry, Antonios Matsakas, Sabine Wenisch, Stefan Arnhold, Ketan Patel
Skeletal muscle mass loss has a broad impact on body performance and physical activity. Muscle wasting occurs due to genetic mutation as in muscular dystrophy, age-related muscle loss (sarcopenia) as well as in chronic wasting disorders as in cancer cachexia. Food restriction reduces muscle mass underpinned by increased muscle protein break down. However the influence of dietary restriction on the morphometry and phenotype of forelimb muscles in a genetically modified myostatin null mice are not fully characterized...
June 13, 2017: Acta Histochemica
https://www.readbyqxmd.com/read/28606474/regulation-of-intracellular-trafficking-and-secretion-of-adiponectin-by-myosin-ii
#9
Deepa Bedi, John C Dennis, Edward E Morrison, Tim D Braden, Robert L Judd
Adiponectin is a protein secreted by white adipocytes that plays an important role in insulin action, energy homeostasis and the development of atherosclerosis. The intracellular localization and trafficking of GLUT4 and leptin in adipocytes has been well studied, but little is known regarding the intracellular trafficking of adiponectin. Recent studies have demonstrated that constitutive adiponectin secretion is dependent on PIP2 levels and the integrity of cortical F-actin. Non-muscle myosin II is an actin-based motor that is associated with membrane vesicles and participates in vesicular trafficking in mammalian cells...
August 19, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28592632/rear-polarized-wnt5a-receptor-actin-myosin-polarity-wramp-structures-promote-the-speed-and-persistence-of-directional-cell-migration
#10
Mary Katherine Connacher, Jian Wei Tay, Natalie G Ahn
In contrast to events at the cell leading edge, rear-polarized mechanisms that control directional cell migration are poorly defined. Previous work described a new intracellular complex, the Wnt5a-receptor-actomyosin polarity (WRAMP) structure, which coordinates the polarized localization of MCAM, actin, and myosin IIB in a Wnt5a-induced manner. However, the polarity and function for the WRAMP structure during cell movement were not determined. Here we characterize WRAMP structures during extended cell migration using live-cell imaging...
July 7, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28538157/cell-and-tissue-scale-forces-coregulate-fgfr2-dependent-tetrads-and-rosettes-in-the-mouse-embryo
#11
Jun Wen, Hirotaka Tao, Kimberly Lau, Haijiao Liu, Craig A Simmons, Yu Sun, Sevan Hopyan
What motivates animal cells to intercalate is a longstanding question that is fundamental to morphogenesis. A basic mode of cell rearrangement involves dynamic multicellular structures called tetrads and rosettes. The contribution of cell-intrinsic and tissue-scale forces to the formation and resolution of these structures remains unclear, especially in vertebrates. Here, we show that Fgfr2 regulates both the formation and resolution of tetrads and rosettes in the mouse embryo, possibly in part by spatially restricting atypical protein kinase C, a negative regulator of non-muscle myosin IIB...
May 23, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/28533420/igf1-stimulates-greater-muscle-hypertrophy-in-the-absence-of-myostatin-in-male-mice
#12
Alexander Hennebry, Jenny Oldham, Tea Shavlakadze, Miranda D Grounds, Philip Sheard, Marta L Fiorotto, Shelley Falconer, Heather K Smith, Carole Berry, Ferenc Jeanplong, Jeremy Bracegirdle, Kenneth Matthews, Gina Nicholas, Mônica Senna-Salerno, Trevor Watson, Christopher D McMahon
Insulin-like growth factors (IGFs) and myostatin have opposing roles in regulating the growth and size of skeletal muscle, with IGF1 stimulating, and myostatin inhibiting, growth. However, it remains unclear whether these proteins have mutually dependent, or independent, roles. To clarify this issue, we crossed myostatin null (Mstn(-/-)) mice with mice overexpressing Igf1 in skeletal muscle (Igf1(+)) to generate six genotypes of male mice; wild type (Mstn(+/+) ), Mstn(+/-), Mstn(-/-), Mstn(+/+):Igf1(+), Mstn(+/-):Igf1(+) and Mstn(-/-):Igf1(+) Overexpression of Igf1 increased the mass of mixed fibre type muscles (e...
August 2017: Journal of Endocrinology
https://www.readbyqxmd.com/read/28475953/effects-of-dietary-starch-types-on-growth-performance-meat-quality-and-myofibre-type-of-finishing-pigs
#13
Y J Li, J L Li, L Zhang, F Gao, G H Zhou
To investigate the effects of dietary starch types on growth performance and meat quality of finishing pigs, ninety barrows (68.0±2.0kg) were randomly allotted to three experimental diets with five replicates of six pigs, containing pure waxy maize starch (WMS), nonwaxy maize starch (NMS) and pea starch (PS) (amylose/amylopectin were 0.07, 0.19 and 0.28 respectively). Compared with WMS diet, PS diet increased the average daily gain, loin eye area, pH45 value, NMR transverse relaxation (T2)2 peak area ratio and sarcoplasmic protein solubility, decreased the feed to gain ratio, back fat, drip loss, cooking loss and T23 peak area ratio (P<0...
May 2, 2017: Meat Science
https://www.readbyqxmd.com/read/28431393/loss-of-tpm4-1-leads-to-disruption-of-cell-cell-adhesions-and-invasive-behavior-in-breast-epithelial-cells-via-increased-rac1-signaling
#14
SukYeong Jeong, SunYoung Lim, Galina Schevzov, Peter W Gunning, David M Helfman
Here we report the identification and characterization of a novel high molecular weight isoform of tropomyosin, Tpm4.1, expressed from the human TPM4 gene. Tpm4.1 expression is down-regulated in a subset of breast cancer cells compared with untransformed MCF10A breast epithelial cells and in highly metastatic breast cancer cell lines derived from poorly metastatic MDA-MD-231 cells. In addition, patients with invasive ductal breast carcinoma show decreased TPM4 expression compared with patients with ductal breast carcinoma in situ, and low TPM4 expression is associated with poor prognosis...
May 16, 2017: Oncotarget
https://www.readbyqxmd.com/read/28352215/myosin-iia-related-actomyosin-contractility-mediates-oxidative-stress-induced-neuronal-apoptosis
#15
Yan Wang, Yingqiong Xu, Qian Liu, Yuanyuan Zhang, Zhen Gao, Mingzhu Yin, Nan Jiang, Guosheng Cao, Boyang Yu, Zhengyu Cao, Junping Kou
Oxidative stress-induced neuronal apoptosis plays an important role in the progression of central nervous system (CNS) diseases. In our study, when neuronal cells were exposed to hydrogen peroxide (H2O2), an exogenous oxidant, cell apoptosis was observed with typical morphological changes including membrane blebbing, neurite retraction and cell contraction. The actomyosin system is considered to be responsible for the morphological changes, but how exactly it regulates oxidative stress-induced neuronal apoptosis and the distinctive functions of different myosin II isoforms remain unclear...
2017: Frontiers in Molecular Neuroscience
https://www.readbyqxmd.com/read/28345668/c9orf135-encodes-a-membrane-protein-whose-expression-is-related-to-pluripotency-in-human-embryonic-stem-cells
#16
Shixin Zhou, Yinan Liu, Yumin Ma, Xiaoyan Zhang, Yang Li, Jinhua Wen
Human embryonic stem cells (hESCs) are a unique population of cells defined by their capacity for self-renewal and pluripotency. Here, we identified a previously uncharacterized gene in hESCs, C9ORF135, which is sharply downregulated during gastrulation and gametogenesis, along with the pluripotency factors OCT4, SOX2, and NANOG. Human ESCs express two C9ORF135 isoforms, the longer of which encodes a membrane-associated protein, as determined by immunostaining and western blotting of fractionated cell lysates...
March 27, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28345221/effect-of-timing-of-electromyostimulation-on-denervated-skeletal-muscle-atrophy
#17
Eun Sil Koh, Hee Chan Kim, Jae-Young Lim
INTRODUCTION: In this study we evaluated the effect of electromyostimulation (EMS) on myosin heavy chain (MHC) isoform expression in denervated rat muscles to determine the optimal timing for EMS application. METHODS: EMS was initiated on post-injury day 1 for the group with denervation receiving immediate EMS (DIEMS) and on post-injury day 15 for the group with denervation receiving delayed EMS (DDEMS) in rat denervated muscles. Muscle wet weight and muscle fiber cross-sectional area (FCSA) were measured...
March 27, 2017: Muscle & Nerve
https://www.readbyqxmd.com/read/28328100/smart-mechanosensing-machineries-enable-migration-of-vascular-smooth-muscle-cells-in-atherosclerosis-relevant-3d-matrices
#18
REVIEW
Qingjia Chi, Jieling Shan, Xiaorong Ding, Tieying Yin, Yazhou Wang, Dongyu Jia, Guixue Wang
At the early stage of atherosclerosis, neointima is formed due to the migration of vascular smooth muscle cells (VSMCs) from the media to the intima. VSMCs are surrounded by highly adhesive 3D matrices. They take specific strategies to cross various 3D matrices in the media, including heterogeneous collagen and mechanically strong basement membrane. Migration of VSMCs is potentially caused by biomechanical mechanism. Most in vitro studies focus on cell migration on 2D substrates in response to biochemical factors...
March 22, 2017: Cell Biology International
https://www.readbyqxmd.com/read/28251924/differential-role-of-nonmuscle-myosin-ii-isoforms-during-blebbing-of-mcf-7-cells
#19
Sumit K Dey, Raman K Singh, Shyamtanu Chattoraj, Shekhar Saha, Alakesh Das, Kankan Bhattacharyya, Kaushik Sengupta, Shamik Sen, Siddhartha S Jana
Bleb formation has been correlated with nonmuscle myosin II (NM-II) activity. Whether three isoforms of NM-II (NM-IIA, -IIB and -IIC) have the same or differential roles in bleb formation is not well understood. Here we report that ectopically expressed, GFP-tagged NM-II isoforms exhibit different types of membrane protrusions, such as multiple blebs, lamellipodia, combinations of both, or absence of any such protrusions in MCF-7 cells. Quantification suggests that 50% of NM-IIA-GFP-, 29% of NM-IIB-GFP-, and 19% of NM-IIC1-GFP-expressing MCF-7 cells show multiple bleb formation, compared with 36% of cells expressing GFP alone...
April 15, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28228547/limch1-regulates-nonmuscle-myosin-ii-activity-and-suppresses-cell-migration
#20
Yu-Hung Lin, Yen-Yi Zhen, Kun-Yi Chien, I-Ching Lee, Wei-Chi Lin, Mei-Yu Chen, Li-Mei Pai
Nonmuscle myosin II (NM-II) is an important motor protein involved in cell migration. Incorporation of NM-II into actin stress fiber provides a traction force to promote actin retrograde flow and focal adhesion assembly. However, the components involved in regulation of NM-II activity are not well understood. Here we identified a novel actin stress fiber-associated protein, LIM and calponin-homology domains 1 (LIMCH1), which regulates NM-II activity. The recruitment of LIMCH1 into contractile stress fibers revealed its localization complementary to actinin-1...
April 15, 2017: Molecular Biology of the Cell
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