scholarly journals Glial imaging during synapse remodeling at the neuromuscular junction

2008 ◽  
Vol 4 (4) ◽  
pp. 319-326 ◽  
Author(s):  
Yi Zuo ◽  
Derron Bishop
Keyword(s):  
Neuromuscular Junction ◽  
Schwann Cells ◽  
Synapse Formation ◽  
Ultrastructural Organization ◽  
Post Injury ◽  
Electron Microscopic ◽  
Cell Dynamics ◽  
Transmission Electron

Glia are an indispensable structural and functional component of the synapse. They modulate synaptic transmission and also play important roles in synapse formation and maintenance. The vertebrate neuromuscular junction (NMJ) is a classic model synapse. Due to its large size, simplicity and accessibility, the NMJ has contributed greatly to our understanding of synapse development and organization. In the past decade, the NMJ has also emerged as an effective model for studying glia–synapse interactions, in part due to the development of various labeling techniques that permit NMJs and associated Schwann cells (the glia at NMJs) to be visualized in vitro and in vivo. These approaches have demonstrated that Schwann cells are actively involved in synapse remodeling both during early development and in post-injury reinnervation. In vivo imaging has also recently been combined with serial section transmission electron microscopic (ssTEM) reconstruction to directly examine the ultrastructural organization of remodeling NMJs. In this review, we focus on the anatomical studies of Schwann cell dynamics and their roles in formation, maturation and remodeling of vertebrate NMJs using the highest temporal and spatial resolution methods currently available.

Morphometrical Ultrastructural Studies of the Rat's Hepatocytes Under Cooling

1997 ◽  
Vol 3 (S2) ◽  
pp. 245-246
Author(s):  
A.S. Kaprelyants ◽  
A.A. Kaprelyants ◽  
A.N. Reylan ◽  
R.K. Migunova
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Electron Microscopic ◽  
Rat Hepatocyte ◽  
Ultrastructural Studies ◽  
Transmission Electron ◽  
Transmission Electron Microscopic ◽  
Morphometric Methods

The aim of given investigation is to study the effect of cooling upon rat hepatocyte structure using transmission electron microscopic and computer morphometric methods. Ultrastructural and morphometrical characteristics of hepatocytes under liver cooling for various levels under in vivo and in vitro conditions were investigated. Vistar rats of 180-250 g were used in the experiment. Liver cooling (in vivo) was performed by means of original cryoapplicator with different probe temperature (1,2). Liver tissue for transmission electron microscopy was fixed in glutaraldehyde fixator on cocadylate buffer and OsO4. Dehydration was completed on acetone (3). Tissue embedding was done into the mixture of Epon/Araldite epoxy rasin. Ultrathin slices were contrasted by the method of Reinolds. Cell viewing and imaging were accomplished by electron microscope at accelerating power of 75kV.Morphometrical and stereometrical analysis was performed using the “Morpho-Tools” original computer system (c) 1994-1996 A.S. Kaprelyants, A.A. Kaprelyants, A.N. Reylan .

scholarly journals Adding exogenous biglycan or decorin improves tendon formation for equine peritenon and tendon proper cells in vitro

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Monica Y. Pechanec ◽  
Tannah N. Boyd ◽  
Keith Baar ◽  
Michael J. Mienaltowski
Keyword(s):  
Material Properties ◽  
Quantitative Polymerase Chain Reaction ◽  
Tendon Repair ◽  
Three Dimensional ◽  
Collagen Content ◽  
Matrix Assembly ◽  
Post Injury ◽  
Transmission Electron

Abstract Background Tendon injuries amount to one of the leading causes of career-ending injuries in horses due to the inability for tendon to completely repair and the high reinjury potential. As a result, novel therapeutics are necessary to improve repair with the goal of decreasing leg lameness and potential reinjury. Small leucine-rich repeat proteoglycans (SLRPs), a class of regulatory molecules responsible for collagen organization and maturation, may be one such therapeutic to improve tendon repair. Before SLRP supplementation can occur in vivo, proper evaluation of the effect of these molecules in vitro needs to be assessed. The objective of this study was to evaluate the effectiveness of purified bovine biglycan or decorin on tendon proper and peritenon cell populations in three-dimensional tendon constructs. Methods Equine tendon proper or peritenon cell seeded fibrin three-dimensional constructs were supplemented with biglycan or decorin at two concentrations (5 nM or 25 nM). The functionality and ultrastructural morphology of the constructs were assessed using biomechanics, collagen content analysis, transmission electron microscopy (TEM), and gene expression by real time – quantitative polymerase chain reaction (RT-qPCR). Results SLRP supplementation affected both tendon proper and peritenon cells-seeded constructs. With additional SLRPs, material and tensile properties of constructs strengthened, though ultrastructural analyses indicated production of similar-sized or smaller fibrils. Overall expression of tendon markers was bolstered more in peritenon cells supplemented with either SLRP, while supplementation of SLRPs to TP cell-derived constructs demonstrated fewer changes in tendon and extracellular matrix markers. Moreover, relative to non-supplemented tendon proper cell-seeded constructs, SLRP supplementation of the peritenon cells showed increases in mechanical strength, material properties, and collagen content. Conclusions The SLRP-supplemented peritenon cells produced constructs with greater mechanical and material properties than tendon proper seeded constructs, as well as increased expression of matrix assembly molecules. These findings provide evidence that SLRPs should be further investigated for their potential to improve tendon formation in engineered grafts or post-injury.

Intraerythrocytic schizogony of Theileria annulata

Parasitology ◽  
1985 ◽  
Vol 91 (1) ◽  
pp. 67-82 ◽  
Author(s):  
P. A. Conrad ◽  
B. G. Kelly ◽  
C. G. D. Brown
Keyword(s):  
Theileria Annulata ◽  
Infected Cattle ◽  
Electron Microscopic ◽  
Trophozoite Stage ◽  
Transmission Electron ◽  
Transmission Electron Microscopic ◽  
Microscopic Studies ◽  
Food Vacuoles

The intraerythrocytic multiplication of two strains of Theileria annulata was studied with parasites maintained in stationary cultures and in the blood of infected cattle. In cultures established with blood from infected cattle 20–60% of single T. annulata piroplasms divided into quadruplet forms by day 6 in vitro. Transmission electron microscopic studies of T. annulata in culture showed that piroplasms possess intracytoplasmic food vacuoles and cytostomes during a pre-division trophozoite stage. The onset of intraerythrocytic multi plication was marked by the appearance of rhoptries and electron-dense plaques beneath the parasite's plasmalemmal membrane. The plaques developed into short segments of subplasmalemmal double membranes which were closely associated with the rhoptries. It was concluded that multiplication of T. annulata in erythrocytes occurred by schizogony, as nuclear division preceded cytoplasmic division and the final separation of merozoites. The four merozoites produced by intraerythrocytic schizogony had the same ultrastructural features as the T. annulata merozoites produced by intralymphocytic schizogony. Clusters of four merozoites, identical to those observed in stationary cultures, were also seen in the erythrocytes of persistently infected cattle and appeared to represent the most significant dividing forms of T. annulata in vivo.

Antiparasitic effect of calcium and magnesium ion-free buffer treatments against a common monogeneanNeobenedenia girellae

Parasitology ◽  
2006 ◽  
Vol 134 (2) ◽  
pp. 229-236 ◽  
Author(s):  
H. OHASHI ◽  
N. UMEDA ◽  
N. HIRAZAWA ◽  
Y. OZAKI ◽  
C. MIURA ◽  
...  
Keyword(s):  
Control Method ◽  
Intercellular Junctions ◽  
Effective Control ◽  
Metallic Ions ◽  
Electron Microscopic ◽  
Percentage Survival ◽  
Transmission Electron ◽  
Spotted Halibut

This study investigated a new effective method for controlling the capsalid monogeneanNeobenedenia girellae. We examinedin vitroandin vivothe effect on the percentage survival ofN. girellaein buffers containing different metallic ions. Decreased survival was observed in buffer solutions lacking two ions. In particular, the percentage survival ofN. girellaewas significantly decreased after 10 min exposure to buffer containing neither Ca2+nor Mg2+. Transmission electron microscopic observations showed that treatment with this buffer disrupted intercellular junctions. This significant effect on percentage survival ofN. girellaeusing Ca2+/Mg2+-free buffer was confirmed in anin vivoassay. Ca2+/Mg2+-free buffer had no effect on the condition of the host, spotted halibutVerasper variegates(Pleuronectidae). These results suggest that treatment with Ca2+/Mg2+-free buffer is a new effective control method, which could replace existing control methods.

Cytological changes induced by platinum-thymine in sarcoma-180 cells: Electron microscopy study

1990 ◽  
Vol 48 (3) ◽  
pp. 290-291
Author(s):  
Gustav Ofosu
Keyword(s):  
Mammalian Cells ◽  
Antitumor Agent ◽  
Microscopy Study ◽  
Electron Microscopy Study ◽  
Limiting Factor ◽  
Sarcoma 180 ◽  
Electron Microscopic ◽  
Cytological Changes

Platinum-thymine has been found to be a potent antitumor agent, which is quite soluble in water, and lack nephrotoxicity as the dose-limiting factor. The drug has been shown to interact with DNA and inhibits DNA, RNA and protein synthesis in mammalian cells in vitro. This investigation was undertaken to elucidate the cytotoxic effects of piatinum-thymine on sarcoma-180 cells in vitro ultrastructurally, Sarcoma-180 tumor bearing mice were treated with intraperitoneal injection of platinum-thymine 40mg/kg. A concentration of 60μg/ml dose of platinum-thymine was used in in vitro experiments. Treatments were at varying time intervals of 3, 7 and 21 days for in vivo experiments, and 30, 60 and 120 min., 6, 12, and 24th in vitro. Controls were not treated with platinum-thymine.Electron microscopic analyses of the treated cells in vivo and in vitro showed drastic cytotoxic effect.

scholarly journals Cemp1-p3 Peptide Promotes the Transformation of Octacalcium Phosphate into Hydroxyapatite Crystals

Crystals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1131
Author(s):  
Maricela Santana ◽  
Gonzalo Montoya ◽  
Raúl Herrera ◽  
Lía Hoz ◽  
Enrique Romo ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Octacalcium Phosphate ◽  
Sequence Motifs ◽  
Simple Method ◽  
Transmission Electron ◽  
Precursor Phase ◽  
Mineralized Tissues ◽  
Potent Growth

Dental cementum contains unique molecules that regulate the mineralization process in vitro and in vivo, such as cementum protein 1 (CEMP1). This protein possesses amino acid sequence motifs like the human recombinant CEMP1 with biological activity. This novel cementum protein 1-derived peptide (CEMP1-p3, from the CEMP1’s N-terminal domain: (QPLPKGCAAVKAEVGIPAPH), consists of 20 amino acids. Hydroxyapatite (HA) crystals could be obtained through the combination of the amorphous precursor phase and macromolecules such as proteins and peptides. We used a simple method to synthesize peptide/hydroxyapatite nanocomposites using OCP and CEMP1-p3. The characterization of the crystals through scanning electron microscopy (SEM), powder X-ray diffraction (XRD), high--resolution transmission electron microscopy (HRTEM), and Raman spectroscopy revealed that CEMP1-p3 transformed OCP into hydroxyapatite (HA) under constant ionic strength and in a buffered solution. CEMP1-p3 binds and highly adsorbs to OCP and is a potent growth stimulator of OCP crystals. CEMP1-p3 fosters the transformation of OCP into HA crystals with crystalline planes (300) and (004) that correspond to the cell of hexagonal HA. Octacalcium phosphate crystals treated with CEMP1-p3 grown in simulated physiological buffer acquired hexagonal arrangement corresponding to HA. These findings provide new insights into the potential application of CEMP1-p3 on possible biomimetic approaches to generate materials for the repair and regeneration of mineralized tissues, or restorative materials in the orthopedic field.

scholarly journals Expression of cytotactin in the normal and regenerating neuromuscular system.

1989 ◽  
Vol 108 (2) ◽  
pp. 625-635 ◽  
Author(s):  
J K Daniloff ◽  
K L Crossin ◽  
M Pinçon-Raymond ◽  
M Murawsky ◽  
F Rieger ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Neuromuscular Junction ◽  
Schwann Cells ◽  
Node Of Ranvier ◽  
Myotendinous Junction ◽  
Neuromuscular System ◽  
Peripheral Nerve Damage ◽  
Neural Cell Adhesion ◽  
Developing Cerebellum

Cytotactin is an extracellular glycoprotein found in a highly specialized distribution during embryonic development. In the brain, it is synthesized by glia, not neurons. It is involved in neuron-glia adhesion in vitro and affects neuronal migration in the developing cerebellum. In an attempt to extend these observations to the peripheral nervous system, we have examined the distribution and localization of cytotactin in different parts of the normal and regenerating neuromuscular system. In the normal neuromuscular system, cytotactin accumulated at critical sites of cell-cell interactions, specifically at the neuromuscular junction and the myotendinous junction, as well at the node of Ranvier (Rieger, F., J. K. Daniloff, M. Pinçon-Raymond, K. L. Crossin, M. Grumet, and G. M. Edelman. 1986. J. Cell Biol. 103:379-391). At the neuromuscular junction, cytotactin was located in terminal nonmyelinating Schwann cells. Cytotactin was also detected near the insertion points of the muscle fibers to tendinous structures in both the proximal and distal endomysial regions of the myotendinous junctions. This was in striking contrast to staining for the neural cell adhesion molecule, N-CAM, which was accumulated near the extreme ends of the muscle fiber. Peripheral nerve damage resulted in modulation of expression of cytotactin in both nerve and muscle, particularly among the interacting tissues during regeneration and reinnervation. In denervated muscle, cytotactin accumulated in interstitial spaces and near the previous synaptic sites. Cytotactin levels were elevated and remained high along the endoneurial tubes and in the perineurium as long as muscle remained denervated. Reinnervation led to a return to normal levels of cytotactin both in inner surfaces of the nerve fascicles and in the perineurium. In dorsal root ganglia, the processes surrounding ganglionic neurons became intensely stained by anticytotactin antibodies after the nerve was cut, and returned to normal by 30 d after injury. These data suggest that local signals between neurons, glia, and supporting cells may regulate cytotactin expression in the neuromuscular system in a fashion coordinate with other cell adhesion molecules. Moreover, innervation may regulate the relative amount and distribution of cytotactin both in muscle and in Schwann cells.

The neural crest population responding to endothelin-3 in vitro includes multipotent cells

1997 ◽  
Vol 110 (14) ◽  
pp. 1673-1682 ◽  
Author(s):  
J.G. Stone ◽  
L.I. Spirling ◽  
M.K. Richardson
Keyword(s):  
Neural Crest ◽  
Schwann Cells ◽  
Cell Types ◽  
Developmental Potential ◽  
Colony Assay ◽  
Cellular Targets ◽  
Multipotent Cells ◽  
Endothelin 3

The peptide endothelin 3 (EDN3) is essential for normal neural crest development in vivo, and is a potent mitogen for quail truncal crest cells in vitro. It is not known which subpopulations of crest cells are targets for this response, although it has been suggested that EDN3 is selective for melanoblasts. In the absence of cell markers for different precursor types in the quail crest, we have characterised EDN3-responsive cell types using in vitro colony assay and clonal analysis. Colonies were analysed for the presence of Schwann cells, melanocytes, adrenergic cells or sensory-like cells. We provide for the first time a description of the temporal pattern of lineage segregation in neural crest cultures. In the absence of exogenous EDN3, crest cells proliferate and then differentiate. Colony assay indicates that in these differentiated cultures few undifferentiated precursors remain and there is a low replating efficiency. By contrast, in the presence of 100 ng/ml EDN3 differentiation is inhibited and most of the cells maintain the ability to give rise to mixed colonies and clones containing neural crest derivatives. A high replating efficiency is maintained. In secondary culture there was a progressive decline in the number of cell types per colony in control medium. This loss of developmental potential was not seen when exogenous EDN3 was present. Cell type analysis suggests two novel cellular targets for EDN3 under these conditions. Contrary to expectations, one is a multipotent precursor whose descendants include melanocytes, adrenergic cells and sensory-like cells; the other can give rise to melanocytes and Schwann cells. Our data do not support previous claims that the action of EDN3 in neural crest culture is selective for cells in the melanocyte lineage.

Neuromuscular target recognition by a homophilic interaction of connectin cell adhesion molecules in Drosophila

Development ◽  
1997 ◽  
Vol 124 (8) ◽  
pp. 1433-1441 ◽  
Author(s):  
A. Nose ◽  
T. Umeda ◽  
M. Takeichi
Keyword(s):  
Cell Adhesion ◽  
Synapse Formation ◽  
Target Recognition ◽  
Surface Protein ◽  
Transgenic Lines ◽  
A Cell ◽  
Neuromuscular Connectivity ◽  
Recognition Molecule

Drosophila Connectin (CON) is a cell surface protein of the leucine-rich repeat family. During the formation of neuromuscular connectivity, CON is expressed on the surface of a subset of embryonic muscles and on the growth cones and axons of the motoneurons that innervate these muscles, including primarily SNa motoneurons and their synaptic targets (lateral muscles). In vitro, CON can mediate homophilic cell adhesion. In this study, we generated transgenic lines that ectopically expressed CON on all muscles. In the transformant embryos and larvae, SNa motoneurons often inappropriately innervated a neighboring non-target muscle (muscle 12) that ectopically expressed CON. Furthermore, the ectopic synapse formation was dependent on the endogenous CON expression on the SNa motoneurons. These results show that CON can function as an attractive and homophilic target recognition molecule in vivo.

Abstract 17071: Cell Communication Inference in Macrophages

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Nika Taghdiri ◽  
Kevin R King ◽  
David Calcagno ◽  
Zhenxing Fu ◽  
Kenneth Huang ◽  
...  
Keyword(s):  
Image Analysis ◽  
Analytical Methods ◽  
Cell Injury ◽  
Diffusion Mechanism ◽  
Cell Communication ◽  
In Vivo Studies ◽  
Cell Dynamics ◽  
Calcium Indicator

Introduction: Tissue macrophages play diverse roles in the cardiovascular system during health and disease. They have diverse functions within tissues, but our understanding of their dynamics is limited because most macrophage characterization assays are destructive and have low temporal resolution. We asked whether these cells are dynamic and interconnected. Methods: Here, we describe experimental and analytical methods for measuring cell dynamics and inferring communication between cells in vitro and in vivo. We created a mouse (Csf1r-Cre x GCaMP5) expressing the Cre-inducible genetically encoded calcium indicator GCaMP5 under the regulation of the innate immune promoter, Csf1r, to non-destructively quantify high-frequency cell dynamics and differentiated them in culture using m-CSF. We developed custom image analysis routines and parameterization strategies for classifying calcium responses. Results: Our studies revealed that calcium reporter BMDMs display minimal fluctuations at baseline but exhibit a dynamic response to immunogenic DNA sensing. DNA-induced isolated cell injury and death, which precipitated cell communication that spread with a velocity of [9μm/s], consistent with an extracellular diffusion mechanism. We developed quantitative image analysis methods that corrected for random calcium fluctuations and identified statistically significant areas of correlated calcium changes suggestive of communication. An analytical pipeline enabled quantification of calcium spike dynamics and correlations of dynamic calcium profiles of single cell sharing a local microenvironment. This resulted in an “improbable synchrony” metric that allowed localization of communication in time and space. We adapted the pipeline for in vivo studies and tested them in a dorsal window chamber model using intravital microscopy. At 2Hz sampling frequency, we identified 27 potential communication events as they responded to complex microenvironmental cues in vivo. Conclusion: The experimental and analytical methods for inferring cell communication provide a new quantitative toolkit for investigating known as-yet undiscovered cell communication pathways.

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