scholarly journals ULTRASTRUCTURE OF SARCOPLASMIC RETICULUM PREPARATIONS

1969 ◽  
Vol 42 (1) ◽  
pp. 296-307 ◽  
Author(s):  
D. W. Deamer ◽  
R. J. Baskin

Fragmented sarcoplasmic reticulum (FSR) from rabbit muscle was examined by positive staining, negative staining, and freeze-etch electron microscopic techniques in the absence and presence of calcium transport conditions. The existence of 30–40 A particles covering the outer surface of FSR vesicles was confirmed by two different negative stains in unfixed, glutaraldehyde-fixed and osmium tetroxide-fixed material. Freeze-etch microscopy revealed a second type of particle, 80–90 A in diameter, on the fractured surfaces of FSR vesicles. Following calcium oxalate accumulation, negative and positive staining techniques provided evidence for large nodular deposits within FSR vesicles which probably correspond to calcium oxalate crystals and are responsible for increments in turbidity during calcium oxalate accumulation. The most probable configuration of FSR vesicles in solution is spherical. "Tadpole" or tubular configurations were not seen by freeze-etch microscopy, positive staining, or in prefixed negatively stained material.

1987 ◽  
Vol 65 (9) ◽  
pp. 1952-1956 ◽  
Author(s):  
J. A. Traquair

Oxalic acid and crystals of calcium oxalate were produced during growth of Leucostoma cincta and L. persoonii on potato dextrose agar and in peach bark tissues. The identification of calcium oxalate was based on solubility characteristics, the results of KMnO4 titration, positive staining with silver nitrate – dithiooxamide, and crystal morphology as observed with light and scanning electron microscopes. Oxalic acid was detected by gas chromatography. This is the first report of oxalic acid production by both Leucostoma species causing peach canker. Calcium oxalate crystals observed on or near hyphae in culture were similar to crystals in artificially inoculated peach bark tissues. Addition of oxalic acid solutions alone to inner bark tissues caused maceration and necrosis. These results indicate a role for oxalic acid in the early stages of pathogenesis by Leucostoma spp. Tetragonal (bipyramidal) and prismatic calcium oxalate crystals formed on bark wounds treated with oxalic acid solutions were similar to those observed in infected tissues and in culture media amended with oxalic acid.


Author(s):  
M. R. Edwards ◽  
W. A. Samsonoff

Papillomas in catfishes have been described (1) but the presence of viruses in these tumors has not (2). This report is concerned with the study of a papilloma found on the lower lip of a brown bullhead (letalurus nebulosus) which had been frozen prior to arrival at our laboratory.Tumor tissues were thawed in 2.5% glutaraldehyde and postfixed in 1$ osmium tetroxide. Both fixatives were prepared in Millonig's buffer. Fixation, washing, dehydration, and Epon embedding were accomplished according to conventional electron microscopic techniques.Examination of thin sections revealed virus-like particles in epidermal cells which constituted the major part of the neoplasm (Fig. 1). No particles were found in the connective tissue surrounding the epidermal papillae. The cells containing particles were usually isolated from one another, had a spindle or fusiform shape, and exhibited many cytoplasmic extensions in random directions. Their nuclei were pleomorphic and displayed irregular nuclear envelopes with relatively large lacunae. Vesiculation of the cytoplasm was extensive, apparently caused by dilations of the endoplasmic reticulum.


1984 ◽  
Vol 62 (10) ◽  
pp. 2028-2032 ◽  
Author(s):  
Z. K. Punja ◽  
S. F. Jenkins

Crystals produced during growth of Sclerotium rolfsii on cellophane overlaying water agar were identified as calcium oxalate based on their solubility characteristics in certain acids, positive staining reaction with silver nitrate – dithiooxamide, and characteristic energy-dispersive X-ray emission spectrum. Addition of calcium salts to water agar enhanced crystal formation. Similar crystals of calcium oxalate were observed with the light and scanning electron microscopes in sugar beet and carrot leaf tissues infected by Sclerotium rolfsii. They frequently formed along the infecting hyphae or were associated with hyphal aggregates and were observed in abundance within the tissue. Addition of oxalic acid solutions to leaf discs resulted in necrosis of the tissue, and crystals similar in all respects to those produced in infected tissues were formed. These observations indicate that the oxalic acid produced by Sclerotium rolfsii in culture or in diseased tissue may sequester available calcium to form calcium oxalate, and provide evidence for the role of oxalic acid in pathogenesis.


1981 ◽  
Vol 195 (1) ◽  
pp. 287-295 ◽  
Author(s):  
K A McGill ◽  
J P Bennett ◽  
G A Smith ◽  
R W Plumb ◽  
G B Warren

A population of sarcoplasmic-reticulum vesicles, all of which were sealed with their cytoplasmic side outwards, was obtained by density-gradient centrifugation after loading the vesicles with calcium oxalate. The calcium oxalate could subsequently be removed from the vesicles by the reverse action of the calcium-transport system. Measurements of the catalysed exchange of the phosphatidylcholine in the sarcoplasmic-reticulum cytoplasmic monolayer with an exogenous phosphatidylcholine pool suggested that phosphatidylcholine is symmetrically distributed across the sarcoplasmic-reticulum membrane. A similar result was obtained for phosphatidylethanolamine when sarcoplasmic-reticulum lipids were labelled with trinitrobenzenesulphonic acid. Further catalysed lipid-exchange reactions showed that the transverse movement of phosphatidylcholine across the membrane was exceedingly slow (t 1/2 greater than 15 days).


Author(s):  
R. A. Waugh ◽  
J. R. Sommer

Cardiac sarcoplasmic reticulum (SR) is a complex system of intracellular tubules that, due to their small size and juxtaposition to such electron-dense structures as mitochondria and myofibrils, are often inconspicuous in conventionally prepared electron microscopic material. This study reports a method with which the SR is selectively “stained” which facilitates visualizationwith the transmission electron microscope.


Author(s):  
Mitsuo Ohtsuki ◽  
Michael Sogard

Structural investigations of biological macromolecules commonly employ CTEM with negative staining techniques. Difficulties in valid image interpretation arise, however, due to problems such as variability in thickness and degree of penetration of the staining agent, noise from the supporting film, and artifacts from defocus phase contrast effects. In order to determine the effects of these variables on biological structure, as seen by the electron microscope, negative stained macromolecules of high density lipoprotein-3 (HDL3) from human serum were analyzed with both CTEM and STEM, and results were then compared with CTEM micrographs of freeze-etched HDL3. In addition, we altered the structure of this molecule by digesting away its phospholipid component with phospholipase A2 and look for consistent changes in structure.


Author(s):  
P. Maupin-Szamier ◽  
T. D. Pollard

We have studied the destruction of rabbit muscle actin filaments by osmium tetroxide (OSO4) to develop methods which will preserve the structure of actin filaments during preparation for transmission electron microscopy.Negatively stained F-actin, which appears as smooth, gently curved filaments in control samples (Fig. 1a), acquire an angular, distorted profile and break into progressively shorter pieces after exposure to OSO4 (Fig. 1b,c). We followed the time course of the reaction with viscometry since it is a simple, quantitative method to assess filament integrity. The difference in rates of decay in viscosity of polymerized actin solutions after the addition of four concentrations of OSO4 is illustrated in Fig. 2. Viscometry indicated that the rate of actin filament destruction is also dependent upon temperature, buffer type, buffer concentration, and pH, and requires the continued presence of OSO4. The conditions most favorable to filament preservation are fixation in a low concentration of OSO4 for a short time at 0°C in 100mM sodium phosphate buffer, pH 6.0.


Author(s):  
W. G. Banfield ◽  
G. Kasnic ◽  
J. H. Blackwell

An ultrastructural study of the intestinal epithelium of mice infected with the agent of epizootic diarrhea of infant mice (EDIM virus) was first performed by Adams and Kraft. We have extended their observations and have found developmental forms of the virus and associated structures not reported by them.Three-day-old NLM strain mice were infected with EDIM virus and killed 48 to 168 hours later. Specimens of bowel were fixed in glutaraldehyde, post fixed in osmium tetroxide and embedded in epon. Sections were stained with uranyl magnesium acetate followed by lead citrate and examined in an updated RCA EMU-3F electron microscope.The cells containing virus particles (infected) are at the tips of the villi and occur throughout the intestine from duodenum through colon. All developmental forms of the virus are present from 48 to 168 hours after infection. Figure 1 is of cells without virus particles and figure 2 is of an infected cell. The nucleus and cytoplasm of the infected cells appear clearer than the cells without virus particles.


Author(s):  
Iracema M. Baccarini

Some morphological nuclear features (invaginations) in normal and abnormal cells have been described in several electron microscopic studies. They have been referred to by others as blebs, loops, pockets, sheets, bodies, nuclear inclusions and cytoplasmic invaginations. Identical appearing structures were found in cells of the uterine cervical epithelium, in trophoblasts of blastocysts and in trophoblasts of rat placenta.Methods. Uterine cervix (normal rats), rat placenta (9-10 days gestation) and blastocyst were placed in 3% glutarahdehyde for 3 hours. The tissue was washed in phosphate buffer for 24 hours, postfixed in 1%. buffered osmium tetroxide for 1-2 hours and embedded in epon araldite. Sections were double stained with uranyl acetate and lead citrate and viewed in E. M. Siemens 200.Observations. Nuclear invaginations were found in basal, parabasal and mucous cells of the cervix epithelium, in trophoblasts of blastocyst and in trophoblasts of placenta. An oval, round or elongated invagination contained heterogenously cytoplasm surrounded by a double intact membrane; usually several invaginations were found in the same nucleus.


Author(s):  
Vicki L. Baliga ◽  
Mary Ellen Counts

Calcium is an important element in the growth and development of plants and one form of calcium is calcium oxalate. Calcium oxalate has been found in leaf seed, stem material plant tissue culture, fungi and lichen using one or more of the following methods—polarized light microscopy (PLM), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and x-ray diffraction.Two methods are presented here for qualitatively estimating calcium oxalate in dried or fixed tobacco (Nicotiana) leaf from different stalk positions using PLM. SEM, coupled with energy dispersive x-ray spectrometry (EDS), and powder x-ray diffraction were used to verify that the crystals observed in the dried leaf with PLM were calcium oxalate.


Sign in / Sign up

Export Citation Format

Share Document