Plasma membrane-associate filament systems in cultured cells visualized by dry-cleaving

1983 ◽  
Vol 64 (1) ◽  
pp. 351-364
Author(s):  
D.A. Mesland ◽  
H. Spiele
Keyword(s):  
Cultured Cells ◽  
Hepatoma Cell ◽  
Cell Types ◽  
Membrane Skeleton ◽  
Adhesive Tape ◽  
Cell Organelles ◽  
Red Cell Membrane ◽  
Transmission Electron ◽  
General Network ◽  
Dry Cleaving

Substrate-attached critical-point-dried cells cleave along the level of the substrate-adherent membrane if removed by means of adhesive tape. The remaining membrane fragments on grids can be visualized three-dimensionally by means of stereo transmission electron microscopy. Attachment of cells may be achieved by active spreading of the cell, or artificially by poly-L-lysine adherence of prefixed cells. In 11 different cell types a filamentous network appears to remain associated with the cytoplasmic face of the membrane. In one hepatoma cell type virtually no filamentous network could be detected. Two general network morphologies are described: the hepatocytic network and the lymphoid network. Since no correspondence could be found between cytoplasmic structure and the structure of the membrane-associated network, and since cells generally cleave along the level of this network, excluding cell organelles, we conclude that it comprises a distinct structural system, analogous to the membrane skeleton of the red cell membrane.

Evaluation of SmartFlare probe applicability for verification of RNAs in early equine conceptuses, equine dermal fibroblast cells and trophoblastic vesicles

2017 ◽  
Vol 29 (11) ◽  
pp. 2157 ◽  
Author(s):  
S. Budik ◽  
W. Tschulenk ◽  
S. Kummer ◽  
I. Walter ◽  
C. Aurich
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Gold Particle ◽  
Developmental Stages ◽  
Cultured Cells ◽  
Dermal Fibroblast ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Limited Information ◽  
Transmission Electron

Live cell RNA imaging has become an important tool for studying RNA localisation, dynamics and regulation in cultured cells. Limited information is available using these methods in more complex biological systems, such as conceptuses at different developmental stages. So far most of the approaches rely on microinjection of synthetic constructs into oocytes during or before fertilisation. Recently, a new generation of RNA-specific probes has been developed, the so named SmartFlare probes (Merck Millipore). These consist of a central 15-nm gold particle with target-specific DNAs immobilised on its surface. Because of their central gold particle, SmartFlare probes are detectable by transmission electron microscopy. The aim of the present study was to investigate the uptake and distribution of SmartFlare probes in equine conceptuses at developmental stages suitable for embryo transfer (Days 6–10), equine trophoblast vesicles and equine dermal fibroblast cell cultures, and to determine whether differences among these cell types and structures exist. Probe uptake was followed by transmission electron microscopy and fluorescence microscopy. Although the embryonic zona pellucida did not reduce uptake of the probe, the acellular capsule fully inhibited probe internalisation. Nanogold particles were taken up by endocytosis by all cell types examined in a similar manner with regard to time and intracellular migration. They were processed in endosomal compartments and accumulated within lysosomal structures after longer incubation times. In conclusion, the SmartFlare probe is applicable in equine conceptuses, but its use is limited to the developmental stages before the formation of the embryonic capsule.

Sequestrocytes: a manifestation of transcellular cross-bonding of the red cell membrane in sickle cell anemia

1989 ◽  
Vol 94 (3) ◽  
pp. 593-600
Author(s):  
R.S. Weinstein ◽  
J.A. Warth ◽  
K. Near ◽  
Y. Marikovsky
Keyword(s):  
Electron Microscopy ◽  
Cell Membrane ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Membrane Skeleton ◽  
Red Cell ◽  
Membrane Injury ◽  
Red Cell Membrane ◽  
Transmission Electron ◽  
Scanning Electron

An unusual form of red cells (called sequestrocytes) that circulate in the peripheral blood of patients with sickle cell anemia has been identified. In wetmount light microscopy preparations, sequestrocytes appear as massively vacuolated erythrocytes. We have shown by transmission electron microscopy and scanning electron microscopy that the morphology of sequestrocytes can be accounted for on the basis of transcellular cross-bonding of the cell membrane. Initially, endocytotic vesicles span the cytoplasmic space, forming putative cytoskeletal fusion zones. Points of fusion of the membrane skeleton bridging the cytoplasmic compartment expand laterally to form linear fusion zones that entrap lakes of hemoglobin within pseudovacuoles. Sequestrocytes can be isolated in the densest layer (1.159 g ml-1 or greater) of an arabinogalactan density gradient. These cells can be generated in increased numbers in sickle cell blood by incubating samples in 1.5 mM-acetylphenylhydrazine (APH) solution for two hours at 37 degrees C. Their formation is partially blocked by incubation with the reducing agent, dithiothreitol (DTT). Our results suggest that these cells represent an expression of oxidative membrane injury in sickle cell anemia.

Cell Reactivity Pattern of the Guinea Pig Cecal Mucosa Evidenced by the ZIO Technique

1982 ◽  
Vol 40 ◽  
pp. 50-51
Author(s):  
Juan Mora-Galindo ◽  
Jorge Arauz-Contreras
Keyword(s):  
Guinea Pig ◽  
Phase Contrast ◽  
Osmium Tetroxide ◽  
Cell Types ◽  
Cell Reactivity ◽  
Transmission Electron ◽  
Neural Tissues ◽  
Maturation Stages ◽  
Zinc Iodide ◽  
Cecal Mucosa

The zinc iodide-osmium tetroxide (ZIO) technique is presently employed to study both, neural and non neural tissues. Precipitates depends on cell types and possibly cell metabol ism as well.Guinea pig cecal mucosa, already known to be composed of epithelium with cells at different maturation stages and lamina propria which i s formed by morphologically and functionally heterogeneous cell population, was studied to determine the pat tern of ZIO impregnation. For this, adult Guinea pg cecal mucosa was fixed with buffered 1.2 5% g 1 utara 1 dehyde before incubation with ZIO for 16 hours, a t 4°C in the dark. Further steps involved a quick sample dehydration in graded ethanols, embedding in Epon 812 and sectioning to observe the unstained material under a phase contrast light microscope (LM) and a transmission electron microscope (TEM).

scholarly journals Morphological and Ultrastructural Characterization of Hemocytes in an Insect Model, the Hematophagous Dipetalogaster maxima (Hemiptera: Reduviidae)

Insects ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 640
Author(s):  
Natalia R. Moyetta ◽  
Fabián O. Ramos ◽  
Jimena Leyria ◽  
Lilián E. Canavoso ◽  
Leonardo L. Fruttero
Keyword(s):  
Cell Biology ◽  
Cell Types ◽  
Transmission Electron ◽  
Ultrastructural Characterization ◽  
Current Classification ◽  
Contrast Microscopy ◽  
First Time ◽  
Controversial Aspect

Hemocytes, the cells present in the hemolymph of insects and other invertebrates, perform several physiological functions, including innate immunity. The current classification of hemocyte types is based mostly on morphological features; however, divergences have emerged among specialists in triatomines, the insect vectors of Chagas’ disease (Hemiptera: Reduviidae). Here, we have combined technical approaches in order to characterize the hemocytes from fifth instar nymphs of the triatomine Dipetalogaster maxima. Moreover, in this work we describe, for the first time, the ultrastructural features of D. maxima hemocytes. Using phase contrast microscopy of fresh preparations, five hemocyte populations were identified and further characterized by immunofluorescence, flow cytometry and transmission electron microscopy. The plasmatocytes and the granulocytes were the most abundant cell types, although prohemocytes, adipohemocytes and oenocytes were also found. This work sheds light on a controversial aspect of triatomine cell biology and physiology setting the basis for future in-depth studies directed to address hemocyte classification using non-microscopy-based markers.

Applying Probe Electrospray Ionization Mass Spectrometry to Cytological Diagnosis: A Preliminary Study by Using Cultured Lung Cancer Cells

2021 ◽  
pp. 1-10
Author(s):  
Yuki Morimoto ◽  
Takeshi Oya ◽  
Mayuko Ichimura-Shimizu ◽  
Minoru Matsumoto ◽  
Hirohisa Ogawa ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Electrospray Ionization ◽  
Squamous Cell ◽  
Cultured Cells ◽  
Sample Pretreatment ◽  
Cell Types ◽  
Diagnostic Tools ◽  
Ionization Mass ◽  
Diagnostic Modality ◽  
Probe Electrospray Ionization

<b><i>Objectives:</i></b> Cytology and histology are 2 indispensable diagnostic tools for cancer diagnosis, which are rapidly increasing in importance with aging populations. We applied mass spectrometry (MS) as a rapid approach for swiftly acquiring nonmorphological information of interested cells. Conventional MS, which primarily rely on promoting ionization by pre-applying a matrix to cells, has the drawback of time-consuming both on data acquisition and analysis. As an emerging method, probe electrospray ionization-MS (PESI-MS) with a dedicated probe is capable to pierce sample and measure specimen in small amounts, either liquid or solid, without the requirement for sample pretreatment. Furthermore, PESI-MS is timesaving compared to the conventional MS. Herein, we investigated the capability of PESI-MS to characterize the cell types derived from the respiratory tract of human tissues. <b><i>Study Design:</i></b> PESI-MS analyses with DPiMS-2020 were performed on various type of cultured cells including 5 lung squamous cell carcinomas, 5 lung adenocarcinomas, 5 small-cell carcinomas, 4 malignant mesotheliomas, and 2 normal controls. <b><i>Results:</i></b> Several characteristic peaks were detected at around m/z 200 and 800 that were common in all samples. As expected, partial least squares-discriminant analysis of PESI-MS data distinguished the cancer cell types from normal control cells. Moreover, distinct clusters divided squamous cell carcinoma from adenocarcinoma. <b><i>Conclusion:</i></b> PESI-MS presented a promising potential as a novel diagnostic modality for swiftly acquiring specific cytological information.

Ultrastructure of the female reproductive organs (ovary, vitellaria, and Mehlis' gland) of Halipegus eccentricus (Trematoda: Derogenidae)

1986 ◽  
Vol 64 (10) ◽  
pp. 2203-2212 ◽  
Author(s):  
Jon M. Holy ◽  
Darwin D. Wittrock
Keyword(s):  
Cell Types ◽  
Reproductive Organs ◽  
Golgi Body ◽  
Digenetic Trematode ◽  
Secretory Cells ◽  
Cortical Granules ◽  
Golgi Bodies ◽  
Transmission Electron ◽  
Vitelline Cells ◽  
Female Reproductive Organs

The female reproductive organs (ovary, vitellaria, and Mehlis' gland) of the digenetic trematode Halipegus eccentricus were studied by transmission electron microscopy. Oocytes entered diplotene while in the ovary and produced cortical granules and lipid bodies. Vitelline cells produced large amounts of eggshell protein but no yolk bodies. Two types of Mehlis' gland secretory cells were present, distinguishable by the morphology of their rough endoplasmic reticulum, Golgi bodies, and secretory bodies, and by the persistence of recognizable secretory material within the ootype lumen after exocytosis. In an attempt to standardize the nomenclature regarding the cell types of the Mehlis' gland, a classification that takes into account these four criteria is proposed. Two basic types of Golgi body organization were noted for the cells of the female reproductive system: a stack of flattened cisternae (Mehlis' gland alpha cells) and spherical Golgi bodies with vesicular cisternae (oocytes, vitelline cells, and Mehlis' gland beta cells).

Fine structure of the kinetochores in six species of the Coleoptera

Genome ◽  
1997 ◽  
Vol 40 (3) ◽  
pp. 379-385
Author(s):  
Klaus Werner Wolf
Keyword(s):  
Direct Contact ◽  
Karyotype Evolution ◽  
Coccinella Septempunctata ◽  
Cell Types ◽  
Male Meiosis ◽  
Serial Sections ◽  
Cell Type ◽  
Present Survey ◽  
Transmission Electron ◽  
Size Variability

Kinetochore structure was examined in a total of 6 species from 5 different families of the Coleoptera using transmission electron microscopy of ultrathin serial sections. Metaphase spermatogonia and primary and secondary spermatocytes were studied in Tenebrio molitor (Tenebrionidae) to determine whether kinetochore structure varies depending on the cell type. In all three cell types, the kinetochore microtubules (MTs) were in direct contact with the chromosomal surface, and kinetochore plates were not detectable. In the other species, only metaphase I spermatocytes were examined. As in T. molitor, distinct kinetochore plates were also absent in Adelocera murina (Elateridae), Agapanthia villosoviridescens (Cerambycidae), and Coccinella septempunctata (Coccinellidae). However, bivalents in male meiosis of two representatives of the Chrysomelidae, Agelastica alni and Chrysolina graminis, showed roughly spherical kinetochores at their poleward surfaces. Microtubules were in contact with this material. Thus, although the present survey covers only a small number of species, it is clear that at least two kinetochore types occur in the Coleoptera. The cytological findings are discussed in the context of chromosome number and genome size variability in the Coleopteran families studied. It is suggested that properties of the kinetochores could play a role in karyotype evolution in the Coleoptera.Key words: bivalent, microtubule, meiosis, metaphase, spermatocyte.

Quantitative Studies of Fibronectin Adsorption on Submicron Scaffolds

2012 ◽  
Author(s):  
Shawn Regis ◽  
Manisha Jassal ◽  
Sina Youssefian ◽  
Nima Rahbar ◽  
Sankha Bhowmick
Keyword(s):  
Surface Functionalization ◽  
Cultured Cells ◽  
Cell Attachment ◽  
Md Simulations ◽  
Cell Types ◽  
Biological Processes ◽  
Integrin Receptors ◽  
Tissue Engineering Scaffolds ◽  
Quantitative Studies

Fibronectin plays a crucial role in adhesion of several cell types, mainly due to the fact that it is recognized by at least ten different integrin receptors. Since most cell types can bind to fibronectin, it becomes involved in many various biological processes. The interaction of cells with ECM proteins such as fibronectin provides the signals affecting morphology, motility, gene expression, and survival of cells [1]. Fibronectin exists in both soluble and insoluble forms; soluble fibronectin is secreted by cells and exits in cell media or body fluids, whereas insoluble fibronectin exists in tissues or the extracellular matrix of cultured cells [2]. The ability to control adsorption of fibronectin on tissue engineering scaffolds would therefore play a huge role in controlling cell attachment and survival in vivo. This can be achieved through surface functionalization of the scaffolds. The goal of these studies is to use molecular dynamics (MD) simulations to mechanistically understand how fibronectin adsorption is enhanced by surface functionalization of submicron scaffolds.

Analysis of a tissue-specific enhancer: ARF6 regulates adipogenic gene expression

1992 ◽  
Vol 12 (3) ◽  
pp. 1202-1208
Author(s):  
R A Graves ◽  
P Tontonoz ◽  
B M Spiegelman
Keyword(s):  
Gene Expression ◽  
Cultured Cells ◽  
Mutational Analysis ◽  
Cell Types ◽  
Specific Gene ◽  
Enhancer Activity ◽  
Cis Acting ◽  
Specific Gene Expression ◽  
Nuclear Extracts ◽  
Adipogenic Gene

The molecular basis of adipocyte-specific gene expression is not well understood. We have previously identified a 518-bp enhancer from the adipocyte P2 gene that stimulates adipose-specific gene expression in both cultured cells and transgenic mice. In this analysis of the enhancer, we have defined and characterized a 122-bp DNA fragment that directs differentiation-dependent gene expression in cultured preadipocytes and adipocytes. Several cis-acting elements have been identified and shown by mutational analysis to be important for full enhancer activity. One pair of sequences, ARE2 and ARE4, binds a nuclear factor (ARF2) present in extracts derived from many cell types. Multiple copies of these elements stimulate gene expression from a minimal promoter in preadipocytes, adipocytes, and several other cultured cell lines. A second pair of elements, ARE6 and ARE7, binds a separate factor (ARF6) that is detected only in nuclear extracts derived from adipocytes. The ability of multimers of ARE6 or ARE7 to stimulate promoter activity is strictly adipocyte specific. Mutations in the ARE6 sequence greatly reduce the activity of the 518-bp enhancer. These data demonstrate that several cis- and trans-acting components contribute to the activity of the adipocyte P2 enhancer and suggest that ARF6, a novel differentiation-dependent factor, may be a key regulator of adipogenic gene expression.

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