scholarly journals Sugammadex affects GH/GHR’s signaling transduction on muscle cells by regulating the membrane-localized GHR level

2022 ◽  
Vol 0 (0) ◽  
Author(s):  
Xiaoqian Lyu ◽  
Fei Xie ◽  
Yujuan Tao ◽  
Jie Bai
Keyword(s):  
Signaling Pathway ◽  
Intracellular Signaling ◽  
Laser Scanning ◽  
Neuromuscular Block ◽  
C2c12 Cell ◽  
Muscle Cells ◽  
Janus Kinase ◽  
Confocal Laser ◽  
Signaling Transduction ◽  
The Relationship

Abstract Objectives Sugammadex (also known as bridion) is a modified γ-cyclodextrin, which is a reversal agent for the neuromuscular block. Growth hormone (GH) has an important biological effect on muscle, regulating muscle growth and development. In the current work, we explored the effect of Sugammadex on GH’s bioactivities. Methods Confocal laser scanning microscope (CLSM), flow cytometry, indirect immunofluorescence, Western-blot, and IP-WB were used to explore the effect of Sugammadex on GH’s bioactivities. Results We found that Sugammadex reduced the activity of GH on muscle cells, which down-regulated GH/GHR-mediated intracellular signaling pathway, such as Janus kinase 2 (JAK2) and signal transducers and activators of transcription 5 (STAT5). We further study the potential biological mechanism by which Sugammadex down-regulated GH/GHR-mediated signaling pathway, a series of related experiments were conducted, and found that Sugammadex may inhibit the proliferation of C2C12 cell via regulating the membrane-localized GHR, which may be the underlying mechanism by which Sugammadex suppressed GHR-induced signaling transduction. This work has laid the theoretical and experimental basis for further exploring the relationship between Sugammadex and GH’s activity. Conclusions In conclusion, this study laid a foundation for further study on the relationship between Sugammadex and GH’s activity.

Altered Cytoskeleton in Smooth Muscle of Aganglionic Bowel

2002 ◽  
Vol 126 (6) ◽  
pp. 692-696
Author(s):  
Laszlo Nemeth ◽  
Udo Rolle ◽  
Prem Puri
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Laser Scanning ◽  
Hirschsprung Disease ◽  
Muscle Cells ◽  
Cytoskeletal Proteins ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Normal Bowel ◽  
Pull Through

Abstract Context.—Intestinal motility is under the control of smooth muscle cells, enteric plexus, and hormonal factors. In Hirschsprung disease (HD), the aganglionic colon remains spastic or tonically enhanced and unable to relax. The smooth muscle cell's cytoskeleton consists of proteins or structures whose primary function is to link or connect protein filaments to each other or to the anchoring sites. Dystrophin is a subsarcolemmal protein with a double adhesion property, one between the membrane elements and the contractile filaments of the cytoskeleton and the other between the cytoskeletal proteins and the extracellular matrix. Desmin and vinculin are functionally related proteins that are present in the membrane-associated dense bodies in the sarcolemma of the smooth muscle cells. Objective.—To examine the distribution of the cytoskeletal proteins in the smooth muscle of the aganglionic bowel. Design.—Bowel specimens from ganglionic and aganglionic sections of the colon were collected at the time of pull-through surgery from 8 patients with HD. Colon specimens collected from 4 patients at the time of bladder augmentation acted as controls. Anti-dystrophin, anti-desmin, and anti-vinculin antibodies were used for fluorescein immunostaining using confocal laser scanning microscopy. Results.—Moderate to strong dystrophin immunoreactivity was observed at the periphery of smooth muscle fibers in normal bowel and ganglionic bowel from patients with HD, whereas dystrophin immunoreactivity was either absent or weak in the smooth muscle of aganglionic colon. Moderate to strong cytoplasmic immunostaining for vinculin and desmin was seen in the smooth muscle of normal bowel and ganglionic bowel from patients with HD, whereas vinculin and desmin staining in the aganglionic colon was absent or weak. Conclusion.—This study demonstrates that the cytoskeletal proteins are abundant in the smooth muscle of normal bowel, but are absent or markedly reduced in the aganglionic bowel of HD. As cytoskeletal proteins are required for the coordinated contraction of muscle cells, their absence may be responsible for the motility dysfunction in the aganglionic segment.

scholarly journals A New Approach to Three-dimensional Reconstructed Imaging of Hormone-secreting Cells and Their Microvessel Environments in Rat Pituitary Glands by Confocal Laser Scanning Microscopy

2000 ◽  
Vol 48 (4) ◽  
pp. 569-577 ◽  
Author(s):  
J. Itoh ◽  
K. Kawai ◽  
A. Serizawa ◽  
K. Yasumura ◽  
K. Ogawa ◽  
...  
Keyword(s):  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Endocrine Cells ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser ◽  
Rat Pituitary ◽  
Pituitary Glands ◽  
The Relationship

There has been considerable interest in the relationship between hormone-secreting endocrine cells and their microvessels in human pituitary gland. However, microcirculatory networks have rarely been studied in three dimensions (3D). This study was designed to visualize and to reveal the relationship between hormone-secreting endocrine cells and their microvessel environment in 3D, using rat pituitary glands under various (hyper/hypo) experimental conditions by confocal laser scanning microscopy (CLSM). Female adult Wistar rats were used after bilateral adrenalectomy or ACTH administration for 2 weeks. Clear 3D reconstructed images of ACTH cells, the microvessel network and counterstained nuclei were obtained at a maximal focus depth of 1 mm by CLSM without any background noise. In the hyperfunctional state, slender cytoplasmic processes of hypertrophic stellate ACTH cells frequently extended to the microvessels. In the hypofunctional state, ACTH cells appeared atrophic and round with scanty cytoplasm, and cytoplasmic adhesions to microvessel network patterns were inconspicuous. Therefore, 3D reconstructed imaging by CLSM is a useful technique with which to investigate the microvessel environment of hormone-secreting cells and has the potential to reveal dynamic hormone-secreting pathways.

A confocal laser scanning microscope designed for indicators with ultraviolet excitation wavelengths

1994 ◽  
Vol 266 (1) ◽  
pp. C303-C310 ◽  
Author(s):  
E. Niggli ◽  
D. W. Piston ◽  
M. S. Kirby ◽  
H. Cheng ◽  
D. R. Sandison ◽  
...  
Keyword(s):  
Heart Muscle ◽  
Laser Scanning ◽  
Muscle Cells ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Scanning Microscope ◽  
Dual Emission ◽  
Laser Scanning Microscope ◽  
Heart Muscle Cells

In this paper we describe the modifications necessary to upgrade, at affordable cost, a commercially available confocal laser scanning microscope for use with ultraviolet (UV) excitation. The optical problems associated with these modifications are described in detail, and easy solutions to solve them are suggested. The optical resolution of the instrument was tested with fluorescent beads and was found to be close to diffraction limited. The light losses due to lateral chromatic aberration were assessed in a thick fluorescent specimen and were found to be comparable to those usually observed with visible light. For a more visual example of the resolution of this instrument, isolated ventricular heart muscle cells were loaded with the fluorescent Ca2+ indicator indo 1. This allowed us to visualize subcellular structural detail and to illustrate the optical sectioning capability of the UV confocal microscope when recording indo 1 emission. Dual-emission line scans were used to perform ratiometric time-resolved detection of Ca2+ transients in voltage-clamped heart muscle cells loaded with the salt form of indo 1. The system presented in this paper should significantly broaden the range of fluorescent indicators that can be used in confocal microscopy.

Signal transduction pathways of IL-1β-mediated iNOS in pulmonary vascular smooth muscle cells

2001 ◽  
Vol 281 (4) ◽  
pp. L816-L823 ◽  
Author(s):  
Jonathan D. Finder ◽  
Jennifer L. Petrus ◽  
Andrew Hamilton ◽  
Raphael T. Villavicencio ◽  
Bruce R. Pitt ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase ◽  
Pulmonary Artery ◽  
Smooth Muscle Cells ◽  
Intracellular Signaling ◽  
Muscle Cells ◽  
Dominant Negative ◽  
Janus Kinase ◽  
Dominant Negative Mutant ◽  
Pulmonary Artery Smooth Muscle

Interleukin (IL)-1β is an important early mediator of inflammation in pulmonary artery smooth muscle cells. We previously reported that a geranylgeranyltransferase inhibitor elevated basal levels of inducible nitric oxide synthase (iNOS) and enhanced IL-1β-mediated induction, suggesting that Rac or Rho small G proteins are candidates for antagonism of such induction. In this study, overexpression of constitutively active Rac1 or its dominant negative mutant did not affect IL-1β induction of iNOS. Alternatively, treatment with Clostridium botulinum C3 exoenzyme, which ADP-ribosylates Rho, was associated with superinduction of iNOS, suggesting an inhibitory role for Rho. IL-1β activated the three mitogen-activated protein kinase (extracellular signal-regulated kinases 1 and 2, c-Jun NH2-terminal kinase/stress-activated protein kinase, and p38) and the Janus kinase (JAK)-signal transducer and activator of transcription pathways. The former two pathways were not associated with IL-1β-mediated iNOS induction, whereas the latter two appeared to have inhibitory roles in iNOS expression. These data suggest that a broad intracellular signaling response to IL-1β in rat pulmonary artery smooth muscle cells results in elevated levels of iNOS that is opposed by the geranylgeranylated small G protein Rho as well as the p38 and JAK2 pathways.

Shrinkage of Tracheid Cells With Desorption Visualized by Confocal Laser Scanning Microscopy

IAWA Journal ◽  
2007 ◽  
Vol 28 (1) ◽  
pp. 29-37 ◽  
Author(s):  
Hiroki Sakagami ◽  
Junji Matsumura ◽  
Kazuyuki Oda
Keyword(s):  
Confocal Laser Scanning Microscopy ◽  
Cell Shape ◽  
Laser Scanning ◽  
Pinus Densiflora ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser ◽  
Environment Chamber ◽  
The Relationship

Confocal laser scanning microscopy (CLSM) was applied as a new method of visualizing the shrinkage of wood and its anisotropy. Control of relative humidity and temperature in a specialized environment chamber made it possible to acquire transverse images of tracheids of Akamatsu (Pinus densiflora) from the saturated condition to the dried condition. The shrinkage of tracheid cells was also determined by measuring the tangential diameter of tracheid and lumen, the radial diameter of tracheid and lumen, and the thickness of tangential and radial walls. Moreover, this technique makes it possible to discuss the relationship between moisture content and tracheid cell shape. We found the CLSM technique to be an effective method for visualizing shrinkage of tracheid cells with desorption.

Prostaglandin E2receptor subtype EP2- and EP4-regulated gene expression profiling in human ciliary smooth muscle cells

2010 ◽  
Vol 42 (3) ◽  
pp. 348-360 ◽  
Author(s):  
Armin Reitmair ◽  
Nils W. G. Lambrecht ◽  
Iskandar Yakubov ◽  
Amelia Nieves ◽  
David Old ◽  
...  
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Signaling Pathway ◽  
Intracellular Signaling ◽  
Regulatory Networks ◽  
Muscle Cells ◽  
Receptor Subtypes ◽  
Aqueous Humor Outflow ◽  
Regulated Gene Expression

Prostanoids are an important class of intraocular pressure (IOP)-lowering antiglaucoma agents that act primarily via increased uveo-scleral aqueous humor outflow through the ciliary body. We have developed two novel PGE2analogs that are specific agonists for the PGE2receptor subtypes EP2 and EP4, respectively. To identify gene regulatory networks and key players that mediate the physiological effects observed in vivo, we performed genomewide expression studies using human ciliary smooth muscle cells. Quantitative real-time RT-PCR confirmed a largely overlapping gene expression profile subsequent to EP2 and EP4 agonist treatment, with 65 significantly regulated genes identified overall, 5 being specific for the EP2 agonist and 6 specific for the EP4 agonist. We found predicted functional cAMP-response elements in promoter regions of a large fraction of the predominantly upregulated genes, which suggests that the cAMP signaling pathway is the most important intracellular signaling pathway for these agonists in these cells. Several target genes were identified that, as part of complex regulatory networks, are implicated in tissue remodeling processes and osmoregulation (e.g., AREG, LOXL3, BMP2, AQP3) and thus may help elucidate the mechanism of action of these IOP-lowering drugs involving the uveo-scleral outflow path.

scholarly journals OP0141 HIGH DIMENSIONAL ANALYSIS REVEAL A NETWORK OF CERTAIN TRANSCRIPTION FACTORS THAT LINK VASCULOPATHY AND ORGAN FIBROSIS IN SYSTEMIC SCLEROSIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 90.2-91
Author(s):  
C. G. Anchang ◽  
B. Matalobos Lawaree ◽  
S. Weber ◽  
S. Rauber ◽  
T. Wohlfahrt ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Laser Scanning ◽  
Muscle Cells ◽  
Laser Scanning Microscopy ◽  
Tube Length ◽  
Confocal Laser ◽  
Research Support ◽  
Organ Fibrosis

Background:Since vascular manifestations such as Raynaud’s phenomenon often precede the onset of other clinical manifestations of systemic sclerosis (SSc), the identification of pathways linking vasculopathy to organ fibrosis might thus provide important insights into early disease mechanisms and allow early targeted intervention for both fibrotic and vascular events.Objectives:In this study we performed high dimensional (HD) analyses to identify mediators that link vasculopathy to organ fibrosis.Methods:HD techniques including RNA-seq, ChIP-seq, ATAC-seq and FISH-seq have been performed to identify mediators in vessels and fibrotic lesions of human skin samples of SSc patients and healthy volunteers. In addition, murine skin and lung tissue samples were analyzed by multi-channel immunofluorescence (IF) and confocal laser scanning microscopy. Microvascular endothelial cells, smooth muscle cells and fibroblasts have been further processed to address their functional attributes with regard to their proliferative, migratory and chemotactic capacity. In vivo models and ex vivomouse fetal metatarsal assays were performed to study fibrotic and angiogenic processes.Results:Bioinformatic HD analyses revealed the ETS transcription factor PU.1 as molecular checkpoint of a network of factors that drive matrix production and fibrotic imprinting in SSc. Within this network ATF3 was significantly upregulated in fibroblasts of skin biopsies of SSc patients and of various organs of fibrosis models. ATF3 deficiency ameliorated fibrosis in various mouse models. Notably, ATF3 was significantly upregulated in vascular cells of fibrotic tissues of SSc patients. Multi-channel IF and confocal laser scanning microscopy of skin and lung biopsies of SSc patients revealed an increased expression of ATF3 especially in microvascular endothelial cells and smooth muscle cells. ATF3 overexpression in smooth muscle cells led to an extensively enhanced proliferation and increased migratory capacity whereas endothelial cells showed a SSc-like phenotype with reduced proliferation and migration. After ATF3 overexpression, tube formation capacity was completely altered as assessed by cumulative tube length, tube numbers and capillary sprouting. To investigate vessel outgrowth from a different perspective, we used theex vivofetal mouse metatarsal assay. ATF3 knockout mice showed a completely altered angiogenic response as assessed by tube length, number of branches and number junctions compared to wildtype controls.Conclusion:We identified PU.1 and ATF3 as key factors in disturbed vasculature and endogenous activated fibroblasts suggesting this axis as a potential therapeutic target intervening both fibrotic and vascular manifestations.Disclosure of Interests:Charles Gwellem Anchang: None declared, Bettina Matalobos Lawaree: None declared, Stefanie Weber: None declared, Simon Rauber: None declared, Thomas Wohlfahrt: None declared, Markus Luber: None declared, Alexander Kreuter: None declared, Georg Schett Speakers bureau: AbbVie, BMS, Celgene, Janssen, Eli Lilly, Novartis, Roche and UCB, Jörg Distler Grant/research support from: Boehringer Ingelheim, Consultant of: Boehringer Ingelheim, Paid instructor for: Boehringer Ingelheim, Speakers bureau: Boehringer Ingelheim, Andreas Ramming Grant/research support from: Pfizer, Novartis, Consultant of: Boehringer Ingelheim, Novartis, Gilead, Pfizer, Speakers bureau: Boehringer Ingelheim, Roche, Janssen

scholarly journals Embryological development of the freshwater crab Esanthelphusa nani (Naiyanetr, 1984) (Brachyura: Gecarcinucidae) using confocal laser scanning microscopy

2020 ◽  
Vol 40 (2) ◽  
pp. 162-171
Author(s):  
Rachata Maneein ◽  
Camille Martinand-Mari ◽  
Julien Claude ◽  
Jirarach Kitana ◽  
Noppadon Kitana
Keyword(s):  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser ◽  
Embryological Development ◽  
The Relationship ◽  
Evolutionary And Developmental Biology ◽  
Potential Area

Abstract We investigated the embryological development of Esanthelphusa nani (Naiyanetr, 1984), a common rice-field crab in northern Thailand, using confocal laser scanning microscopy. The development of E. nani can be completed in eggs within 12 d resulting in a hatchling stage corresponding to the megalopa stage. Pre-organogenetic stages were characterized by a superficial cleavage, including egg cleavage, egg blastula, and egg gastrula. Organogenesis stages were identified by the appearances of appendages and was divided into egg nauplius, egg zoea, and egg megalopa. Crabs metamorphose to juveniles after hatching. By providing new comparative data, our study shed some new light on the relationship between environment, phylogeny, and development, opening a potential area of research from the perspective of ecological, evolutionary, and developmental biology.

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