scholarly journals The ultrastructural investigation of mitochondria in B-CLL cells during apoptosis

2004 ◽  
Vol 12 (3) ◽  
pp. 139-141 ◽  
Author(s):  
Goran Brajuskovic ◽  
Andjelija Skaro-Milic ◽  
Slobodan Marjanovic ◽  
Snezana Cerovic ◽  
Slavica Knezevic-Usaj
Keyword(s):  
Close Association ◽  
Critical Role ◽  
Morphological Characteristics ◽  
Uranyl Acetate ◽  
Lymphocytic Leukemia ◽  
Mitochondrial Morphology ◽  
Thin Sections ◽  
Ultrastructural Investigation ◽  
Human Malignancy ◽  
Cacodylate Buffer

BACKGROUND: B-chronic lymphocytic leukemia (B-CLL) is an example of human malignancy caused by alternations in the pathways of apoptosis. Mitochondria play a critical role in the regulation of this process. The B-CLL cells dying in apoptosis showed typical morphological characteristics: the reduction of the nuclear volume is accompanied with the reduction of the cytoplasmatic volume, while many of organelles remain intact. The aim of our study was ultrastructural investigation of mitochondrial morphology in apoptotic B- CLL cells. METHODS: Our study included peripheral blood samples from 32 B-CLL patients. The samples were fixed in 4% glutar-aldehyde buffered in 0.1 cacodylate buffer and postfixed in 1% osmium tetroxide in the same buffer. The specimens were dehydrated in a graded series of alcohol and embedded in EPON 812. The ultra-thin sections were stained with uranyl acetate and lead citrate. Ultrastructural analysis of sections was performed on Philips electron microscope 208S at 80 kV. RESULTS: The most frequent mitochondrial abnormalities in apoptotic B-CLL cells were a reduction of size with a hyperdensity of their matrix (mitochondrial pyknosis), or markedly swollen mitochondria with peripherally placed, disorientated, and disintegrated cristae. In some apoptotic cells, we also detected close association of mitochondria with loops of rough endoplasmatic reticulum. CONCLUSION: The results of our study showed the numerous of mitochondria damages in B-CLL cells during apoptotic process. The correlation between ultrastructural damage and functional activity of mitochondria in apoptotic B-CLL cells is still not clear and requires further investigation.

The Nature of Rapidly Extracted Phycocyanin from the Blue-Green Alga Plectonema Boryanum

1971 ◽  
Vol 29 ◽  
pp. 366-367
Author(s):  
M. Kessel ◽  
R. MacColl
Keyword(s):  
Self Assembly ◽  
Analytical Ultracentrifugation ◽  
Uranyl Acetate ◽  
The Self ◽  
Major Protein ◽  
Subunit Structure ◽  
Blue Green Alga ◽  
Thin Sections ◽  
Blue Green Algae ◽  
Cacodylate Buffer

The major protein of the blue-green algae is the biliprotein, C-phycocyanin (Amax = 620 nm), which is presumed to exist in the cell in the form of distinct aggregates called phycobilisomes. The self-assembly of C-phycocyanin from monomer to hexamer has been extensively studied, but the proposed next step in the assembly of a phycobilisome, the formation of 19s subunits, is completely unknown. We have used electron microscopy and analytical ultracentrifugation in combination with a method for rapid and gentle extraction of phycocyanin to study its subunit structure and assembly.To establish the existence of phycobilisomes, cells of P. boryanum in the log phase of growth, growing at a light intensity of 200 foot candles, were fixed in 2% glutaraldehyde in 0.1M cacodylate buffer, pH 7.0, for 3 hours at 4°C. The cells were post-fixed in 1% OsO4 in the same buffer overnight. Material was stained for 1 hour in uranyl acetate (1%), dehydrated and embedded in araldite and examined in thin sections.

Membrane-bound vesicles associated with the microvilli in the midgut of the freshwater microcrustacean, Daphnia magna

1983 ◽  
Vol 41 ◽  
pp. 480-481
Author(s):  
Patricia L. Jansma
Keyword(s):  
Daphnia Magna ◽  
Intestinal Epithelial Cells ◽  
Uranyl Acetate ◽  
Intestinal Epithelial ◽  
Chlamydomonas Reinhardii ◽  
Thin Sections ◽  
Saturated Water ◽  
Cacodylate Buffer ◽  
Membrane Bound ◽  
Ethanol Series

The presence of the membrane bound vesicles or blebs on the intestinal epithelial cells has been demonstrated in a variety of vertebrates such as chicks, piglets, hamsters, and humans. The only invertebrates shown to have these microvillar blebs are two species of f1ies. While investigating the digestive processes of the freshwater microcrustacean, Daphnia magna, the presence of these microvillar blebs was noticed.Daphnia magna fed in a suspension of axenically grown green alga, Chlamydomonas reinhardii for one hour were narcotized with CO2 saturated water. The intestinal tracts were excised in 2% glutaraldehyde in 0.2 M cacodyl ate buffer and then placed in fresh 2% glutaraldehyde for one hour. After rinsing in 0.1 M cacodylate buffer, the sample was postfixed in 2% OsO4, dehydrated with a graded ethanol series, infiltrated and embedded with Epon-Araldite. Thin sections were stained with uranyl acetate and Reynolds lead citrate before viewing with the Philips EM 200.

Gap junction associated filaments in testis interstitial cells of the rat

1978 ◽  
Vol 36 (2) ◽  
pp. 550-551
Author(s):  
J.F. David-Ferreira ◽  
K.L. David-Ferreira ◽  
M.H. Miranda ◽  
J.C. Lemos
Keyword(s):  
Gap Junction ◽  
Freeze Fracture ◽  
Interstitial Cells ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Thin Sections ◽  
Cacodylate Buffer ◽  
Old Rats ◽  
The Relationship ◽  
Made In

The testis interstitial cells of the rat are closely associated forming small aggregates in the vicinity of the vessels. The finality of the present study is to analise the relationship between the interstitial cells and to describe some particularities of their junctions.The observations were made in thin sections from testis of 2 to 3 months old rats fixed by immersion or perfusion with 2 to 3% glutaral- dehyde in s-collidine or cacodylate buffer followed by 2% osmium tetro-xyde in the same buffers. Some specimens have been prepared following the technique of Shea. The thin sections obtained after embedding in Epon were double stained in uranyl acetate and lead citrate. For the cryofracture study the tissues were fixed in 3% glutaraldehyde in cacodylate buffer, impregnated in 28% glycerol, frozen in Freon 22 and stored in liquid nitrogen. Freeze fracture and platinum-carbon shadowing were done in a Balzers 300.

Ultrastructure of E. coli: An in vitro study of cells cultured in the presence of four gastrointestinal (GI) hormones

1990 ◽  
Vol 48 (3) ◽  
pp. 628-629
Author(s):  
Leo J. Henz ◽  
Frank E. Johnson
Keyword(s):  
Morphological Changes ◽  
In Vitro Study ◽  
Uranyl Acetate ◽  
Culture Collection ◽  
En Bloc ◽  
Thin Sections ◽  
E Coli ◽  
Cacodylate Buffer ◽  
Synthetic Hormones

Hormones are found in exocrine secretions entering the gut. They alter the morphology of many eukaryotic cells; whether they affect the morphology of enteric flora is unknown. In this study, we examined the ultrastructure of E. coli, a common bacterium in the mammalian gut, for morphological changes resulting from exposure to GI hormones.E. coli (#11775 from American Type Culture Collection) were grown in protease-free trypticase soy broth (TSB) at 37°C for 18 hr to a concentration of 2 x 107 cells/ml. Pure synthetic hormones were used: sulfated C-terminal cholecystokinin octapeptide (CCK), pentagastrin (PG), cyclic somatostatin tetradecapeptide (SS), or the porcine form of secretin (SEC). These were individually added to. bacterial cultures in TSB to make 1 x 107 organisms/ml and 0.0, 0.5, 2.5, or 5.0 μg of hormone/ml, then incubated for 30 min at 37°C. The cultures were rapidly chilled and added to equal volumes of cold 6% glutaraldehyde in 0.2 M cacodylate buffer. After 30 min, the bacteria were concentrated by centrifugation (15 min at 4000 RPM) and the pellets suspended in cold 3% glutaraldehyde for an additional 15 min, followed by centrifugation. The pellets were resuspended in cold cacodylate buffer and stored at 2°C for 1-7 d. The cells were again centrifuged and the pellets were blotted with a strip of filter paper to remove excess fluid, then mixed with a drop of warm 2% agar. The agar suspensions were pipetted into cold saline. The resulting solidified extrusions were cut by hand into 2 mm segments for further processing in 1% OsO4 (with or without en bloc staining in 2% uranyl acetate (UA) in ethanol). Following dehydration in ethanol, rinsing in propylene oxide, and encapsulation in Epon-Araldite, thin sections were examined and photographed with a JEOL-100C microscope.

Ultrastructural Characteristics of Vaginal Epithelium of Persistent Estrous Rats

1985 ◽  
Vol 43 ◽  
pp. 658-659
Author(s):  
Khosho Francis K. ◽  
Kaufmann Robert C. ◽  
Amankwah Kofi S.
Keyword(s):  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Ruthenium Red ◽  
Female Rats ◽  
Constant Light ◽  
Ultrastructural Changes ◽  
Vaginal Smears ◽  
Thin Sections ◽  
Cacodylate Buffer ◽  
Ultrastructural Characteristics

Adult female rats exposed to constant light will develop anovulatory acyclicity characterized by persistent vaginal cornification (PE) and formation of multiple large cystic follicles on the ovaries. The purpose of the present communication is to describe the ultrastructural changes in vaginal epithelia in PE rats as compared to that in normal estrous rats.Persistent vaginal estrous with PCO was induced in a group of Sprague-Dawely rats by exposure to constant light for 50-150 days. Rats in normal estrous, as determined by vaginal smears, were used as controls. Nembutal- anethesized rats were perfused through the aorta with 2.5% gluteraldehyde in 1M sodium cacodylate buffer (pH 7.3). The mucosa of the vaginal folds just inferior to the cervix were dissected by microsurgery, postfixed, stained with 0.5% ruthenium red in 1% osmium tetroxide, dehydrated, and embedded in polybed. Thick sections (1μ) were stained with toludine blue for light microscopy studies. Thin sections were stained with uranyl acetate and lead citrate.

Ultrastructural Study of Trophozoites and Cysts of Acanthamoeba-Hartmannella and Naegleria Species

1974 ◽  
Vol 32 ◽  
pp. 192-193
Author(s):  
A. Julio Martinez ◽  
Richard J. Duma ◽  
Doris G. Fultz ◽  
Ruth B. Finley
Keyword(s):  
Present Report ◽  
Uranyl Acetate ◽  
Osmic Acid ◽  
Acetate Solution ◽  
Lead Citrate ◽  
Thin Sections ◽  
Cacodylate Buffer ◽  
In Situ Fixation ◽  
Diamond Knife

Ultrastructural components of trophozoites and cysts of Acanthamoeba-Hartmannella (A-H) and Naegleria amebas have been investigated. The present report describes a technique to study the morphological features of such protozoa.The A-H strains selected were cultured for 7 days in axenic amebic media, and the Naegleria strains were cultured for 7 days in axenic media with blood. In situ fixation was accomplished by adding an equal volume of 4% Osmic Acid in Cacodylate buffer at 4¶C for one hour. Following frequent agitation, the tubes were then spun at 2500 RPM for 10 minutes to obtain a pellet. The pellets were dehydrated in graded series of alcohols (from 50% to absolute) and propylene oxide and then embedded in EPON. One micron sections were stained with Paragon (PS 1301) for thirty seconds. Thin sections were cut with a diamond knife, double stained with 5% alcoholic uranyl acetate solution and lead citrate and observed in an Hitachi-HS-8F2 electron microscope.

The Role of the Internal Coating in the Ultrastructural Architecture of Pinocytic Vesicles and its Significance in Vesicular Transport

1971 ◽  
Vol 29 ◽  
pp. 398-399
Author(s):  
T. Shirahama ◽  
A. S. Cohen ◽  
O. G. Rodgers
Keyword(s):  
Body Weight ◽  
Uranyl Acetate ◽  
Ruthenium Red ◽  
Lead Citrate ◽  
Healthy Young Adult ◽  
Thin Sections ◽  
Cacodylate Buffer ◽  
New Zealand White Rabbits ◽  
Pinocytic Vesicles

Forty-micron nonfrozen sections of myocardium from several healthy young adult New Zealand white rabbits, without pretreatment or 10-15 minutes after an intravenous ferritin injection (100 mg ferritin/100 gm. body weight), were double fixed with 2% formaldehyde - 2.5% glutaraldehyde in 0.1M cacodylate buffer and 2% OSO4 in 0.1M cacodylate buffer in the presence of 1000, 100, 50 or 20 ppm ruthenium red (RR), and embedded in Epon. Thin sections of small blood vessels including capillaries, unstained or stained with uranyl acetate and/or lead citrate, were photographed at initial magnifications of 40,000 or 80,000. The results were analysed to delineate the ultrastructural relation of mucopolysaccharides (MPS) in the architecture of pinocytic vesicles and the MPS role in pinocytosis.

Redistribution of Membranes during Conidiogeneses and Nuclear Development in Verticillium Dahliae

1977 ◽  
Vol 35 ◽  
pp. 344-345
Author(s):  
J. E. Mayfield ◽  
W. J. Tolmsoff ◽  
M. H. Wheeler
Keyword(s):  
Present Report ◽  
Nuclear Material ◽  
Uranyl Acetate ◽  
Thin Sections ◽  
Cacodylate Buffer ◽  
Primary Means ◽  
Nuclear Development ◽  
Single Nucleus ◽  
Conidial Formation ◽  
And Migration

Verticil1ium dahliae is an asexual fungus that produces unicellular, mononucleate conidia as its primary means of reproduction. For conidial formation, specialized hyphal cells convert into verticillate conidiophores, bearing tapered phialides. A conidium develops from the phialide as a protrusion through the terminal pore. Each phialide has a single nucleus that divides repeatedly during the sequential production of numerous conidia. Nuclear material has been found to migrate into developing conidia just before their separation from the phialide tip (1). The present report suggests a possible mechanism for nuclear division and migration.The ultrastructure of phialides and developing conidia of V. dahliae was examined from a narrow zone 5 mm behind the periphery of colonies grown on potato-carrot-dextrose agar. Cells were fixed with 3l glutaraldehyde in cacodylate buffer (pH 6.8) at room temperature, postfixed in cold 2% OsO4 in the same buffer, and thin sections were counter stained with 2% uranyl acetate and lead citrate.

Ultrastructural Observations on Microspore Development in Rice Anther Culture

1985 ◽  
Vol 43 ◽  
pp. 646-647
Author(s):  
S. S. Ren ◽  
J. C. Chen ◽  
Y. R. Chen
Keyword(s):  
Anther Culture ◽  
Oryza Sativa L ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Microspore Development ◽  
Thin Sections ◽  
Cacodylate Buffer ◽  
Fine Structures ◽  
Development In Vitro

The plants produced from rice anther culture varied in ploidy level. Genetic analysis of anther-derived plants indicated genome multiplication occurred in microspore development in vitro. Cytological studies on early roicrospore division suggested endoreduplication and nuclear fusion may be related to the production of nonhaploid plants. In the present study, fine structures of callus ontogeny in anther culture were emphasized.Anthers of rice(Oryza sativa L. c. v. Hsinchu 4, 2n=24) containing mid-uninucleate microspores were excised and cultured in Ng liquid medium, supplimented with 60 g/l sucrose, 1 mg/l kinetin and 2 mg/l naphthalenacetic acid. Cultured anthers were collected at 2 days intervals for 20 days and fixed in 2.5% glutaraldehyde in 0.1M cacodylate buffer(pH=7.0), postfixed in osmium tetraoxide, and then embedded in Spurr's resin. Thin sections were stained with uranyl acetate and lead citrate, observed under Hitachi H-600 at 75 KV.

Nucleolus-like bodies (Nuages) and annulate lamellae in spermatogonia of fish-tilapia, Oreochromis niloticus

1990 ◽  
Vol 48 (3) ◽  
pp. 92-93
Author(s):  
T. Guha ◽  
P.F. Prentis
Keyword(s):  
Electron Microscopy ◽  
Oreochromis Niloticus ◽  
Osmium Tetroxide ◽  
Type A ◽  
Uranyl Acetate ◽  
Annulate Lamellae ◽  
Thin Sections ◽  
Cacodylate Buffer ◽  
Guppy Fish ◽  
Type A Spermatogonia

Type A spermatogonia in tilapia (Oreochromis ni1oticus) have been studied by electron microscopy. These are stem cells from which spermatogenesis beings in this species. In this paper we report presence of two cytoplasmic organelles, annulate lamellae and nucleolus-like bodies (nuages), in type A spermatogonia in O. niloticus.Testes were fixed in 2% gluteraldehyde for 4 hrs. at 4°C and then in 1% osmium tetroxide for 1 hr. at 4°C, both in 0.1M cacodylate buffer (pH 7.4). Fixed tissues were processed in the conventional way for electron microscopy. Thin sections of tissues were stained by uranyl acetate and lead citrate. These were examined in a Carl Zeiss electron microscope operated at 40kV.Nucleolus-like bodies (nuages) have been reported in rat spermatocytes, early postimplantation rat embryos, fish and amphibian oocytes and guppy (fish) spermatogonia. Annulate lamellae have been found in fish spermatogonia and oocytes. Type A spermatogonia (Figs. 1,2,3,4) in O. niloticus show presence of nucleolus-like bodies (nuages) and annulate lamellae in the cytoplasm.

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