Abstract:
:We have used plant root tips frozen under high pressure in conjunction with freeze-fracture electron microscopy a) to evaluate the quality of freezing of unfixed, non-cryoprotected tissues obtainable with this method, b) to examine the structure of cells frozen under high pressure, c) to evaluate the usefulness of high pressure freezing to preserve transient membrane events, and d) to look for artifacts caused by the high pressure. A single artifact of high pressure, possibly related to the collapse of air spaces during pressurization before freezing, manifested itself as long tears or folds in the plasma membrane. Excellent freezing, as evidenced by the smooth, turgid appearance of all membrane systems and the lack of aggregated cytosolic materials was observed in 10 to 20% of samples. In the best preserved specimens freezing was uniform throughout the sample volume and all organelles were readily identified. In the remaining ones, a gradient of ice crystal sizes was seen; cells within 50 to 100 microns of the surface being better preserved than those in the interior. Cortical microtubules appeared well preserved as were close associations of endoplasmic reticulum (ER) with nuclear, Golgi and plasma membranes. Junctions between the ER and nuclear membrane were constricted and much thinner (30 nm in diameter) than in chemically-fixed, thin-sectioned tissue, and although no continuities between the ER and Golgi membranes were observed, many Golgi stacks had an adjacent ER cisterna either at the cis or trans face. Both Golgi and ER cisternae exhibited distinct, round dilations indicative of vesicle blebbing or vesicle fusion events. Characteristic disc- and horseshoe-shaped infoldings of the plasma membrane corresponding to fused secretory vesicle and/or membrane recycling structures were also prominent in many cells. Short extensions of the cortical ER cisternae were regularly observed appressed against these plasma membrane infoldings suggesting a functional role for the ER in vesicle-mediated secretion and/or membrane recycling. Many lipid bodies were intimately associated with the ER, some with their surface monolayer fused with the cytoplasmic leaflet of the ER membrane. Our findings demonstrate that high pressure freezing can provide excellent morphological preservation of intact tissues and can preserve fast, transient membrane events such as those associated with vesicle fusion and vesicle blebbing. We conclude that this is the best available method for freezing relatively large (up to 0.6 mm thick) tissue samples for study by electron microscopy.
journal_name
Eur J Cell Bioljournal_title
European journal of cell biologyauthors
Craig S,Staehelin LAsubject
Has Abstractpub_date
1988-04-01 00:00:00pages
81-93issue
1eissn
0171-9335issn
1618-1298journal_volume
46pub_type
杂志文章abstract::Adoptive T cell therapy is the transfer of T cells to a patient in order to combat disease. This procedure is mainly being used but not limited to the treatment of viral infections and malignancies including virus-associated tumors. Depending on the clinical context, the T cell donor may be the same patient or another...
journal_title:European journal of cell biology
pub_type: 杂志文章,评审
doi:10.1016/j.ejcb.2011.04.001
更新日期:2012-01-01 00:00:00
abstract::The isolation and characterization of highly purified and structurally well-preserved peroxisomes from the renal cortex of different mammalian species (beef, sheep, and cat) is reported. Renal cortex tissue was homogenized and a peroxisome-enriched light mitochondrial fraction was prepared by differential centrifugati...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1986-03-01 00:00:00
abstract::Lipid droplet precursors of milk lipid globules are believed to be derived from elements of endoplasmic reticulum in milk-secreting mammary epithelial cells. Endoplasmic reticulum isolated from mammary gland was able to generate small droplets morphologically resembling microlipid droplet precursors of milk lipid glob...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1992-02-01 00:00:00
abstract::Chondrocyte terminal differentiation is associated with cellular hypertrophy increased activity of plasma membrane alkaline phosphatase and the synthesis of collagen type X. The hypertrophic phenotype of cultured chondrocytes can be stimulated by ascorbic acid but the underlying mechanisms for this phenotypic change a...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/s0171-9335(98)80023-x
更新日期:1998-06-01 00:00:00
abstract::Crude protein extracts of Uromyces appendiculatus contain a polypeptide that resembles actin in several ways. This protein eluates from DEAE-cellulose with concentrations of KCl known to release actin of other species from the cation. The polypeptide is recognized by polyclonal antibodies directed to sodium dodecyl su...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1990-12-01 00:00:00
abstract::The fine structure of a joint receptor (R10) in a spider leg (Zygiella x-notata) was examined with light and electron microscopy. The R10 receptor consists of a compact ganglion which is situated near the dorsal joint membrane of the femur/patella joint. Each of the ten sensory cells comprising the ganglion sends one ...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1979-06-01 00:00:00
abstract::Programmed cell death (PCD) is the regulated removal of cells within an organism and plays a fundamental role in growth and development in nearly all eukaryotes. In animals, the model organism Caenorhabditis elegans (C. elegans) has aided in elucidating many of the pathways involved in the cell death process. Various ...
journal_title:European journal of cell biology
pub_type: 杂志文章,评审
doi:10.1016/j.ejcb.2012.02.002
更新日期:2012-08-01 00:00:00
abstract::Despite its frequent inactivation in human breast cancers, the role of p21(Cip1) (p21) in morphological plasticity of normal mammary epithelial cells is still poorly understood. To address this question, we have investigated the consequences of p21 silencing in two-dimensional (2D) morphogenesis of untransformed human...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/j.ejcb.2011.03.002
更新日期:2011-08-01 00:00:00
abstract::Native tetrameric alpha 2-macroglobulin molecules (alpha 2M) can be converted into a population of dimers by incubation with various divalent cations such as Zn, Cd, Mg, Cu, Ni, Co. This dissociation is completed within 30 min at 37 degrees C. These dimers have a characteristic shape and a size of about 16 X 8 nm, and...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1986-12-01 00:00:00
abstract::Initiation of eukaryotic DNA replication is achieved by the sequential binding of different proteins to origins of DNA replication. Using EGFP-tagged initiator proteins and immunofluorescence techniques we found that most of the ORC and the MCM subunits are localised at centrosomes and are colocalised with the polo-li...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/j.ejcb.2006.09.002
更新日期:2007-01-01 00:00:00
abstract::Rabbit antisera against homogeneous rat liver thioredoxin and thioredoxin reductase (NADPH-oxidized thioredoxin oxidoreductase, E.C. 1.6.4.5) were prepared and used for immunohistochemical analysis in adult rats. Immunoreactive thioredoxin and thioredoxin reductase were widely distributed in tissues and organs, but va...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1985-07-01 00:00:00
abstract::Actin and tubulins of Phytophthora infestans germlings were detected with monoclonal antibodies on Western blots of crude extracts separated by one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The Mr of actin was approximately 43,000, whereas alpha- and beta-tubulin, which migrated...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1990-10-01 00:00:00
abstract::The Semper cells in the ommatidia of the blowfly contain an impressive cytoskeleton consisting of hundreds of microtubules. The microtubules are intimately interwoven by an extensive system of the smooth endoplasmic reticulum. The cells were permeabilized by saponin treatment, subsequently incubated in a loading mediu...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1983-11-01 00:00:00
abstract::MUC16/CA125 is over-expressed in human epithelial tumors including ovarian, breast and some other carcinomas. The purpose of this study is to investigate how cell surface MUC16 is functionally involved in tumor progression, with a special focus on the role of its cytoplasmic tail. Forced expression of C-terminal MUC16...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/j.ejcb.2013.10.005
更新日期:2013-08-01 00:00:00
abstract::The mammalian skin has a photosensitive system comprised by several opsins, including rhodopsin (OPN2) and melanopsin (OPN4). Recently, our group showed that UVA (4.4 kJ/m2) leads to immediate pigment darkening (IPD) in murine normal and malignant melanocytes. We show the role of OPN2 and OPN4 as UVA sensors: UVA-indu...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/j.ejcb.2018.01.004
更新日期:2018-04-01 00:00:00
abstract::We describe here the cloning, expression and characterisation of centaurin-alpha2 from a rat adipocyte cDNA library. The centaurin-alpha2 cDNA contains an open reading frame, which codes for a protein of 376 amino acids with predicted mass of 43.5 kDa. Centaurin-alpha2 shares 51-59% identity with centaurin-alpha1 prot...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1078/0171-9335-00242
更新日期:2002-04-01 00:00:00
abstract::Connexins26 and -32 are subunit proteins of gap junctions that are coexpressed in hepatocytes and several tissues but individually expressed in other cells. Molecular cloning of both corresponding mouse genes revealed similar genomic organization, i.e., each gene consists of two exons with the complete coding region l...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1992-06-01 00:00:00
abstract::A new, two step method is described for the isolation of megakaryocytes from bone marrow. By means of centrifugation in a PercollR density gradient megakaryocytes can be enriched by factor 68 or 24 starting from rat or guinea pig bone marrow, respectively. The purity of the megakaryocytes can be improved ca. 4 fold if...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1980-06-01 00:00:00
abstract::Previously we reported the novel observation that astrocytes ensheath the persistent hyaloid artery, both in the Nuc1 spontaneous mutant rat, and in human PFV (persistent fetal vasculature) disease (Developmental Dynamics 234:36-47, 2005). We now show that astrocytes isolated from both the optic nerve and retina of Nu...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/j.ejcb.2011.01.003
更新日期:2011-05-01 00:00:00
abstract::A zymogen granule fraction has been isolated from rat pancreas, and its purity has been assessed by biochemical and morphological criteria. Specific activities of two marker enzymes, amylase and chymotrypsin, are increased by 4.6 and 5.4-fold, respectively, as compared to the homogenate. The purified fraction is devoi...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1982-08-01 00:00:00
abstract::The interaction of daunomycin (DAU), an anthracyclinic antibiotic employed as antitumoral agent, with microtubules, has been investigated by cytochemical and morphological methods on a human melanoma cell line (H14). Results obtained indicated that DAU was able to modulate the microtubule reassembly in cells treated w...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1991-04-01 00:00:00
abstract::Seminal roots are initiated at the scutellar node during maize (Zea mays L.) embryo development. The maize mutant rtcs (rootless concerning crown and seminal roots) does not initiate seminal roots while its wild-type siblings form on average 2.9 seminal roots per seedling. In this study, proteome profiles of 25-day-ol...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/j.ejcb.2009.11.014
更新日期:2010-02-01 00:00:00
abstract::Coexistence of four different forms of chromatin was observed by electron microscopy in nuclear spread preparations of monkey kidney cells during late stages of infection with herpes simplex virus (HSV-1 AMG). Besides typical nucleosomal (i) chromatin, thin (3-5 nm) strands morphologically indistinguishable from prote...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1980-12-01 00:00:00
abstract::The interaction between myelin basic protein (MBP) and G-actin was studied under nonpolymerizing conditions, i.e.,2mM HEPES, pH 7.5, 0.1 mM CaCl2 and 0.2 mM ATP. Fluorescence studies using pyrenyl-actin and the measurements of ATP hydrolysis rate show that MBP induces changes in the structure of the actin monomer simi...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1986-10-01 00:00:00
abstract::The capacity of vascular endothelial cells to modulate their phenotype in response to changes in environmental conditions is one of the most important characteristics of this cell type. Since different growth factors may play an important signalling role in this adaptive process we have investigated the effect of endo...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1078/0171-9335-00084
更新日期:2000-09-01 00:00:00
abstract::We have investigated the effects of bafilomycin A1, a specific inhibitor of vacuolar type H(+)-ATPase, on the secretion and intracellular distribution of prolactin in cultured rat pituitary tumor cells (GH3 cells). Pulse-labeling the cells with L-[35S]methionine for 5 min and subsequently incubating in chase medium co...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1993-10-01 00:00:00
abstract::Human enteric α-defensins (HD5 and HD6), major antimicrobial peptides produced by Paneth cells in the intestine, play important roles in intestinal innate immunity. Since their expression is decreased in Crohn's disease (CD), with decreased expression being more pronounced in the presence of NOD2 mutations, it would b...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/j.ejcb.2014.10.007
更新日期:2015-01-01 00:00:00
abstract::The present study was undertaken because of the paucity of information on peroxisomes in molluscs and the increasing importance of these organisms as sensitive indicators of environmental pollution. Peroxisomes were identified by electron microscopy in all three main cell types of the digestive gland of the bivalve mo...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1992-12-01 00:00:00
abstract::The transcriptional status of cells can be deduced from the staining pattern of various nuclear markers such as the Cajal body, nucleolus and nuclear speckles. In this study we have used these markers to correlate transcriptional status with cell differentiation in the lens. As a closed system with no cell loss and wi...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1078/0171-9335-00275
更新日期:2002-10-01 00:00:00
abstract::The entry of sporozoites of Theileria parva into bovine lymphoid cells in vitro was studied with the electron microscope. Endocytosis is completed in less than 10 min. No local mobilization of actin or other cytoskeletal elements is detected in the cytoplasm of the cell being invaded and no engulfing pseudopods are fo...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1982-04-01 00:00:00