Ultrastructural Changes in the Root Meristem Cells of Rubus Chamaemorus L. (Cloudberry)

Author(s):  
Arya K. Bal

In the course of studies in the root meristem tissue of Rubus chamaemorus L. some important changes in the ultrastructural morphology were observed during the initiation of senescence at the end of the growing season.Root meristems were collected from naturally growing healthy populations of Cloudberry plants, and fixed in Karnovsky's mixture or in 2.5% glutaraldehyde in phosphate buffer. The samples were osmicated, dehydrated following usual methods and embedded in Epon. Ultrathin sections were stained in uranyl acetate and lead citrate.Figure 1 shows part of a dense cell in the meristem. The electron density of these cells is due to large amounts of a particulate material in the cytoplasmic matrix. The smallest particle seen in electron micrographs is about 40 A, although larger aggregates are also found, which remain randomly distributed in association with various cell organelles. Dense substance has been found associated with golgi membranes, proplastids, vacuoles and microtubules (Fig. 2).

2016 ◽  
Vol 66 (2) ◽  
pp. 245-256 ◽  
Author(s):  
Juraj Pivko ◽  
V. Alexander Makarevich ◽  
Elena Kubovičova ◽  
Jan Rafay ◽  
Peter Chrenek

Abstract The aim of the study was to define ultrastructural changes in the ovarian cyclic corpus luteum (CL) of dairy cows with different body condition score (BCS, a 5-point scale). The ovaries were recovered from slaughtered Holstein cows of BCS1 (emaciation; n= 4), BCS2 (tendency to emaciation; n=4), BCS3 (average body condition; n= 4), BCS4 (tendency to fattening; n=4) and BCS5 (fattening; n=4). Average ages of cows were: 6.2, 4.1, 5.7, 5.5 and 6.36 years for BCS1, BCS2, BCS3, BCS4 and BCS5, respectively. For electron microscopy, functioning CL at the mid-luteal phase were fixed in the aldehyde solution and post-fixed in 1% OsO4 solution. For lipid visualization the samples were soaked in 1% p-phenylenediamine solution in acetone. Durcupan ultrathin sections (90 nm) were contrasted with uranyl acetate and lead citrate and viewed under Jeol transmission electron microscope. Occurrence of cell organelles was determined by counting the relative volume of individual cellular components using the modified universal test network. Luteal regression of cyclic CL was characterized by atrophy of lutein cells, condensation of organelles and cytoplasmic inclusions, increase in relative volumes of vacuoles, lysosomes, peroxisomes, autophagosomes and lipids. There were significant differences (p<0.05) among BCS1, BCS2, BCS4 and BCS5 cows in the occurrence of lipids, vacuoles, lysosomes and autophagosomes in the lutein cells compared to BCS3 cows. Cows with BCS2 or BCS1 had higher relative volume of lipid droplets, whilst the BCS4 or BCS5 cows had increased occurrence (relative volume) of vacuoles, lysosomes, peroxisomes and autolysosomes. These results show that luteolysis of cyclic CL of cows has different patterns in each BCS group; in the emaciated and fattened cows luteolysis starts earlier than in cows with average body condition.


Author(s):  
T. M. Valentine ◽  
T. D. Anderson

Interleukin-2 is being evaluated in cancer immunotherapy in people. rIL-2 is associated with vascular leak syndrome (VLS) characterized by peripheral edema, ascites and pulmonary edema. The histopathologic changes associated with rIL-2 administration have been described in mouse models, however, the ultrastructural changes in VLS are undefined. In these studies we have utilized a mouse model of VLS to characterize the pulmonary ultrastructural changes which are associated with pulmonary edema.B6D2F1 (BDF) mice, (3M/3F/groups) from Charles River Lab., New York, were treated intraperitoneally for five days with vehicle for rIL-2 or with 30,000 U/gm rIL-2, BID as previously described. At necropsy, lungs were fixed by intratracheally at perfusion pressure of 30cm of H2O with modified Karnovskys fixative diluted with 0.2M sodium caccdylate buffer, ph7.4, for 24 hours. Transverse sections of the left lung (1mm thick) were post-fixed in osmium tetroxide (2hrs) and processed for electron microscopy. Ultrathin sections were stained with uranyl acetate and lead citrate and examined in a Philips 300 electron microscope.


Author(s):  
S. K. Aggarwal ◽  
P. McAllister ◽  
R. W. Wagner ◽  
B. Rosenberg

Uranyl acetate has been used as an electron stain for en bloc staining as well as for staining ultrathin sections in conjunction with various lead stains (Fig. 1). Present studies reveal that various platinum compounds also show promise as electron stains. Certain platinum compounds have been shown to be effective anti-tumor agents. Of particular interest are the compounds with either uracil or thymine as one of the ligands (cis-Pt(II)-uracil; cis-Pt(II)-thymine). These compounds are amorphous, highly soluble in water and often exhibit an intense blue coloration. These compounds show enough electron density to be used as stains for electron microscopy. Most of the studies are based on various cell lines (human AV, cells, human lymphoma cells, KB cells, Sarcoma-180 ascites cells, chick fibroblasts and HeLa cells) while studies on tissue blocks are in progress.


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

In this paper we review some of the contributions that electron microscopy has made to the analysis of actin and myosin from nonmuscle cells. We place particular emphasis upon the limitations of the ultrastructural techniques used to study these cytoplasmic contractile proteins, because it is not widely recognized how difficult it is to preserve these elements of the cytoplasmic matrix for electron microscopy. The structure of actin filaments is well preserved for electron microscope observation by negative staining with uranyl acetate (Figure 1). In fact, to a resolution of about 3nm the three-dimensional structure of actin filaments determined by computer image processing of electron micrographs of negatively stained specimens (Moore et al., 1970) is indistinguishable from the structure revealed by X-ray diffraction of living muscle.


Author(s):  
K. Kovacs ◽  
E. Horvath ◽  
J. M. Bilbao ◽  
F. A. Laszlo ◽  
I. Domokos

Electrolytic lesions of the pituitary stalk in rats interrupt adenohypophysial blood flow and result in massive infarction of the anterior lobe. In order to obtain a deeper insight into the morphogenesis of tissue injury and to reveal the sequence of events, a fine structural investigation was undertaken on adenohypophyses of rats at various intervals following destruction of the pituitary stalk.The pituitary stalk was destroyed electrolytically, with a Horsley-Clarke apparatus on 27 male rats of the R-Amsterdam strain, weighing 180-200 g. Thirty minutes, 1,2,4,6 and 24 hours after surgery the animals were perfused with a glutaraldehyde-formalin solution. The skulls were then opened and the pituitary glands removed. The anterior lobes were fixed in glutaraldehyde-formalin solution, postfixed in osmium tetroxide and embedded in Durcupan. Ultrathin sections were stained with uranyl acetate and lead citrate and investigated with a Philips 300 electron microscope.


Author(s):  
Kosuke Ueda ◽  
Hiroto Washida ◽  
Nakazo Watari

IntroductionHemoglobin crystals in the red blood cells were electronmicroscopically reported by Fawcett in the cat myocardium. In the human, Lessin revealed crystal-containing cells in the periphral blood of hemoglobin C disease patients. We found the hemoglobin crystals and its agglutination in the erythrocytes in the renal cortex of the human renal lithiasis, and these patients had no hematological abnormalities or other diseases out of the renal lithiasis. Hemoglobin crystals in the human erythrocytes were confirmed to be the first case in the kidney.Material and MethodsTen cases of the human renal biopsies were performed on the operations of the seven pyelolithotomies and three ureterolithotomies. The each specimens were primarily fixed in cacodylate buffered 3. 0% glutaraldehyde and post fixed in osmic acid, dehydrated in graded concentrations of ethanol, and then embedded in Epon 812. Ultrathin sections, cut on LKB microtome, were doubly stained with uranyl acetate and lead citrate.


Author(s):  
Amankwah K.S. ◽  
A.D. Weberg ◽  
R.C. Kaufmann

Previous research has revealed that passive (involuntary inhalation) tobacco smoking during gestation can have adverse effects upon the developing fetus. These prior investigations did not concentrate on changes in fetal morphology. This study was undertaken to delineate fetal neural abnormalities at the ultrastructural level in mice pups exposed in utero to passive maternal smoking.Pregnant study animals, housed in a special chamber, were subjected to cigarette smoke daily from conception until delivery. Blood tests for determination of carbon monoxide levels were run at 15-18 days gestation. Sciatic nerve tissue from experimental and control animals were obtained following spontaneous delivery and fixed in 2.5% gluteraldehyde in 0.1M cacodylate buffer pH 7.3. The samples were post-fixed in osmium ferrocyanide (1:1 mixture of 1.5% aqueous OSO4 and 2.5% K4 Fe(CN)6). Following dehydration, the tissues were infiltrated with and embedded in Spurr. Sections were stained with uranyl acetate and lead citrate.


Author(s):  
S.L. Asa ◽  
K. Kovacs ◽  
J. M. Bilbao ◽  
R. G. Josse ◽  
K. Kreines

Seven cases of lymphocytic hypophysitis in women have been reported previously in association with various degrees of hypopituitarism. We report two pregnant patients who presented with mass lesions of the sella turcica, clinically mimicking pituitary adenoma. However, pathologic examination revealed extensive infiltration of the anterior pituitary by lymphocytes and plasma cells with destruction of the gland. To our knowledge, the ultrastructural features of lymphocytic hypophysitis have not been studied so far.For transmission electron microscopy, tissue from surgical specimens was fixed in glutaraldehyde, postfixed in OsO4, dehydrated and embedded in epoxy-resin. Ultrathin sections were stained with uranyl acetate and lead citrate and examined with a Philips 300 electron microscope.Electron microscopy revealed adenohypophysial cells of all types exhibiting varying degrees of injury. In the areas of most dense inflammatory cell infiltration pituitary cells contained large lysosomal bodies fusing with secretory granules (Fig. 1), as well as increased numbers of swollen mitochondria, indicating oncocytic transformation (Fig. 2).


Author(s):  
R.H.M. Cross ◽  
C.E.J. Botha ◽  
A.K. Cowan ◽  
B.J. Hartley

Senescence is an ordered degenerative process leading to death of individual cells, organs and organisms. The detection of a conditional lethal mutant (achloroplastic) of Hordeum vulgare has enabled us to investigate ultrastructural changes occurring in leaf tissue during foliar senescence.Examination of the tonoplast structure in six and 14 day-old mutant tissue revealed a progressive degeneration and disappearance of the membrane, apparently starting by day six in the vicinity of the mitochondria associated with the degenerating proplastid (Fig. 1.) where neither of the plastid membrane leaflets is evident (arrows, Fig. 1.). At this stage there was evidence that the mitochondrial membranes were undergoing retrogressive changes, coupled with disorganization of cristae (Fig. 2.). Proplastids (P) lack definitive prolamellar bodies. The cytoplasmic matrix is largely agranular, with few endoplasmic reticulum (ER) cisternae or polyribosomal aggregates. Interestingly, large numbers of actively-budding dictysomes, associated with pinocytotic vesicles, were observed in close proximity to the plasmalemma of mesophyll cells (Fig. 3.). By day 14 however, mesophyll cells showed almost complete breakdown of subcellular organelle structure (Fig. 4.), and further evidence for the breakdown of the tonoplast. The final stage of senescence is characterized by the solubilization of the cell wall due to expression and activity of polygalacturonase and/or cellulose. The presence of dictyosomes with associated pinocytotic vesicles formed from the mature face, in close proximity to both the plasmalemma and the cell wall, would appear to support the model proposed by Christopherson for the secretion of cellulase. This pathway of synthesis is typical for secretory glycoproteins.


Author(s):  
Taber A. Ba-Omar ◽  
Philip F. Prentis

We have recently carried out a study of spermiogenic differentiation in two geographically isolated populations of Aphanius dispar (freshwater teleost), with a view to ascertaining variation at the ultrastructural level. The sampling areas were the Jebel Al Akhdar in the north (Group A) and the Dhofar region (Group B) in the south. Specimens from each group were collected, the testes removed, fixed in Karnovsky solution, post fixed in OsO, en bloc stained with uranyl acetate and then routinely processed to Agar 100 resin, semi and ultrathin sections were prepared for study.


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