scholarly journals Effect of Ca2+ on the dimeric structure of scallop sarcoplasmic reticulum.

1989 ◽  
Vol 108 (2) ◽  
pp. 511-520 ◽  
Author(s):  
L Castellani ◽  
P M Hardwicke ◽  
C Franzini-Armstrong
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Surface Structure ◽  
Calcium Binding ◽  
Freeze Fracture ◽  
Hill Coefficient ◽  
Half Maximum ◽  
Crystalline Surface ◽  
Maximum Activation ◽  
The Individual

Scallop sarcoplasmic reticulum (SR), visualized in situ by freeze-fracture and deep-etching, is characterized by long tubes displaying crystalline arrays of Ca2+-ATPase dimer ribbons, resembling those observed in isolated SR vesicles. The orderly arrangement of the Ca2+-ATPase molecules is well preserved in muscle bundles permeabilized with saponin. Treatment with saponin, however, is not needed to isolate SR vesicles displaying a crystalline surface structure. Omission of ATP from the isolation procedure of SR vesicles does not alter the dimeric organization of the Ca2+-ATPase, although the overall appearance of the tubes seems to be affected: the edges of the vesicles are scalloped and the individual Ca2+-ATPase molecules are not clearly defined. The effect of Ca2+ on isolated scallop SR vesicles was investigated by correlating the enzymatic activity and calcium-binding properties of the Ca2+-ATPase with the surface structure of the vesicles, as revealed by electron microscopy. The dimeric organization of the membrane is preserved at Ca2+ concentrations where the Ca2+ binds to the high affinity sites (half-maximum saturation at pCa approximately 7.0 with a Hill coefficient of 2.1) and the Ca2+-ATPase is activated (half-maximum activation at pCa approximately 6.8 with a Hill coefficient of 1.84). Higher Ca2+ concentrations disrupt the crystalline surface array of the SR tubes, both in the presence and absence of ATP. We discuss here whether the Ca2+-ATPase dimer identified as a structural unit of the SR membrane represents the Ca2+ pump in the membrane.

scholarly journals Morphology of isolated triads.

1983 ◽  
Vol 96 (4) ◽  
pp. 1017-1029 ◽  
Author(s):  
R D Mitchell ◽  
A Saito ◽  
P Palade ◽  
S Fleischer
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Thin Section ◽  
Freeze Fracture ◽  
Structural Features ◽  
Negative Staining ◽  
Transverse Tubule ◽  
Freeze Fracture Electron Microscopy ◽  
Fracture Electron ◽  
Terminal Cisternae

The triad is the junctional association of transverse tubule with sarcoplasmic reticulum terminal cisternae. A procedure for the isolation of highly enriched triads from skeletal muscle has been described in the previous paper. In the present study, the structural features of isolated triads have been examined by thin-section, negative-staining, and freeze-fracture electron microscopy. In isolated triads, key features of the structure observed in situ have been retained, including the osmiophilic "feet," junctional structures between the transverse tubule and terminal cisternae. New insight into triad structure is obtained by negative staining, which also enables visualization of feet at the junctional face of the terminal cisternae, whereas smaller surface particles, characteristic of calcium pump protein, are not visualized there. Therefore, the junctional face is different from the remainder of the sarcoplasmic reticulum membrane. Junctional feet as viewed by thin section or negative staining have similar periodicity and extend approximately 100 A from the surface of the membrane. Freeze-fracture of isolated triads reveals blocklike structures associated with the membrane of the terminal cisternae at the junctional face, interjunctional connections between the terminal cisternae and t-tubule, and intragap particles. The intragap particles can be observed to be closely associated with the t-tubule. The structure of isolated triads is susceptible to osmotic and salt perturbation, and examples are given regarding differential effects on transverse tubules and terminal cisternae. Conditions that adversely affect morphology must be considered in experimentation with triads as well as in their preparation and handling.

scholarly journals Membrane asymmetry and enhanced ultrastructural detail of sarcoplasmic reticulum revealed with use of tannic acid.

1978 ◽  
Vol 79 (3) ◽  
pp. 601-616 ◽  
Author(s):  
A Saito ◽  
C T Wang ◽  
S Fleischer
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Tannic Acid ◽  
Freeze Fracture ◽  
Membrane Vesicles ◽  
Thin Sections ◽  
Membrane Asymmetry ◽  
The Asymmetry ◽  
First Time ◽  
Negative Staining Electron Microscopy

Fixation of purified sarcoplasmic reticulum (SR) membrane vesicles, using glutaraldehyde supplemented with 1% tannic acid, reveals newly visualized ultrastructure in thin sections. The trilaminar appearance of the membrane is highly asymmetric; the outer electron-opaque layer is appreciably wider (70 A) than the inner layer (20 A). The asymmetry is not referable to lack of penetration of the tannic acid since: (a) SR vesicles made permeable with 1 mM EDTA, pH 8.5, show similar asymmetry; (b) treatment of SR with trypsin results in progressive loss in protein content and decrease in the thickness of the outer layer, until in the limit the trilayer has a symmetric appearance; (c) within the same muscle section, the SR membrane appears highly asymmetric whereas the sarcolemma has a more symmetric appearance; (d) reconstituted SR vesicles have a symmetric appearance with equally broad inner and outer layers (approximately 70 A); the symmetric structure is confirmed by freeze-fracture and negative staining electron microscopy. Heavy and light SR vesicles obtained by isopycnic density sedimentation of purified SR have the same asymmetric appearance of the membrane and seem to differ mainly in that the heavy vesicles contain internal contents consisting largely of Ca++-binding protein. The asymmetry of the SR membrane is referable mainly to the unidirectional alignment of the Ca++ pump protein, the major component (90% of the protein) of the membrane. The asymmetry of the SR membrane can be visualized now for the first time in situ in thin sections of muscle.

Electron Probe Analysis of Muscle

1982 ◽  
Vol 40 ◽  
pp. 398-401
Author(s):  
A. V. Somlyo ◽  
H. Shuman ◽  
A. P. Somlyo
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Sarcoplasmic Reticulum ◽  
Resting State ◽  
Binding Sites ◽  
Electron Probe ◽  
Frog Skeletal Muscle ◽  
Electron Probe Analysis ◽  
Probe Analysis

Electron probe analysis of frozen dried cryosections of frog skeletal muscle, rabbit vascular smooth muscle and of isolated, hyperpermeab1 e rabbit cardiac myocytes has been used to determine the composition of the cytoplasm and organelles in the resting state as well as during contraction. The concentration of elements within the organelles reflects the permeabilities of the organelle membranes to the cytoplasmic ions as well as binding sites. The measurements of [Ca] in the sarcoplasmic reticulum (SR) and mitochondria at rest and during contraction, have direct bearing on their role as release and/or storage sites for Ca in situ.

Profile Imaging of Silicon Surface Reconstructions

1985 ◽  
Vol 43 ◽  
pp. 262-263
Author(s):  
O.L. Krivanek ◽  
G.J. Wood
Keyword(s):  
Electron Microscopy ◽  
Surface Structure ◽  
Structure Determination ◽  
Silicon Surface ◽  
Good Resolution ◽  
Surface Reconstructions ◽  
Bulk Structure ◽  
Point To Point ◽  
The Relationship

Electron microscopy at 0.2nm point-to-point resolution, 10-10 torr specimei region vacuum and facilities for in-situ specimen cleaning presents intere; ing possibilities for surface structure determination. Three methods for examining the surfaces are available: reflection (REM), transmission (TEM) and profile imaging. Profile imaging is particularly useful because it giv good resolution perpendicular as well as parallel to the surface, and can therefore be used to determine the relationship between the surface and the bulk structure.

scholarly journals Selected In Situ Hybridization Methods: Principles and Application

Molecules ◽  
2021 ◽  
Vol 26 (13) ◽  
pp. 3874
Author(s):  
Dominika Veselinyová ◽  
Jana Mašlanková ◽  
Katarina Kalinová ◽  
Helena Mičková ◽  
Mária Mareková ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Imaging Techniques ◽  
Cell Communication ◽  
Probe Design ◽  
Rapid Progress ◽  
In Situ Techniques ◽  
Different Types ◽  
Laboratory Procedures ◽  
The Individual

We are experiencing rapid progress in all types of imaging techniques used in the detection of various numbers and types of mutation. In situ hybridization (ISH) is the primary technique for the discovery of mutation agents, which are presented in a variety of cells. The ability of DNA to complementary bind is one of the main principles in every method used in ISH. From the first use of in situ techniques, scientists paid attention to the improvement of the probe design and detection, to enhance the fluorescent signal intensity and inhibition of cross-hybrid presence. This article discusses the individual types and modifications, and is focused on explaining the principles and limitations of ISH division on different types of probes. The article describes a design of probes for individual types of in situ hybridization (ISH), as well as the gradual combination of several laboratory procedures to achieve the highest possible sensitivity and to prevent undesirable events accompanying hybridization. The article also informs about applications of the methodology, in practice and in research, to detect cell to cell communication and principles of gene silencing, process of oncogenesis, and many other unknown processes taking place in organisms at the DNA/RNA level.

scholarly journals In situ detection of vitamin D-induced calcium-binding protein (9-kDa CaBP) messenger RNA in rat duodenum.

1986 ◽  
Vol 34 (2) ◽  
pp. 277-280 ◽  
Author(s):  
M Warembourg ◽  
O Tranchant ◽  
C Perret ◽  
C Desplan ◽  
M Thomasset
Keyword(s):  
Vitamin D ◽  
In Situ Hybridization ◽  
Calcium Binding ◽  
Binding Protein ◽  
Cdna Probe ◽  
Calcium Binding Protein ◽  
Pbr322 Dna ◽  
Rat Duodenum ◽  
Columnar Cells

We have previously described the molecular cloning of a cDNA fragment synthesized from rat duodenal mRNA coding for a 9000-dalton vitamin D-induced calcium-binding protein (9-kDa CaBP) (3). We now report the use of this cloned cDNA to study the cytological distribution of 9-kDa CaBP mRNA in rat duodenum by in situ hybridization. Tissue sections, fixed in ethanol:acetic acid, were hybridized to the 3H-cDNA probe and processed for autoradiography. The specificity of the CaBP mRNA-DNA hybrid formation was checked using 3H-labeled plasmid pBR322 DNA as a control probe. 9k-Da CaBP mRNA, visualized by silver grains, was found only in the absorptive epithelial cells, and the concentration was greater in the cells at the villous tips than in those of the crypts. The 9k-Da CaBP mRNA was observed mainly in the cytoplasm of the columnar cells and less frequently in the nucleus. Labeling was not seen in the brush border and goblet cells. The submucosa, with Brunner's glands and muscularis, also showed no specific 9-kDa CaBP mRNA concentration. This demonstration of 9-kDa CaBP gene activity in the columnar cells of the rat duodenum illustrates the usefulness of in situ hybridization for characterization of specific cells involved in the expression of 1,25(OH)2 D3 activity.

scholarly journals In-Situ Tribology Study of Nanoprobe Sliding on Defective Crystalline Surface

2009 ◽  
Vol 15 (S2) ◽  
pp. 734-735
Author(s):  
Y Liao ◽  
LD Marks
Keyword(s):  
Crystalline Surface ◽  
In Situ Tribology

Extended abstract of a paper presented at Microscopy and Microanalysis 2009 in Richmond, Virginia, USA, July 26 – July 30, 2009

The Effect of Homoeologous Meiotic Pairing in Tetraploid Hordeum bulbosum L. × H. vulgare L. Hybrids on Alien Introgressions in Offspring

2016 ◽  
Vol 150 (2) ◽  
pp. 139-149 ◽  
Author(s):  
Margret Scholz ◽  
Galina Pendinen
Keyword(s):  
Interspecific Hybrids ◽  
In Situ Hybridisation ◽  
Basic Material ◽  
Meiotic Pairing ◽  
Hordeum Bulbosum ◽  
Homoeologous Pairing ◽  
Introgression Breeding ◽  
The Individual ◽  
Pairing Behaviour

The pairing behaviour of the individual chromosome arms of Hordeum vulgare (Hv) with their homoeologous arms of H. bulbosum (Hb) at metaphase I of meiosis in tetraploid Hb × Hv hybrids and the frequencies of recombined Hv chromosome arms in selfed offspring were studied on differentially visualized chromosomes after fluorescent in situ hybridisation. The frequencies of paired Hv-Hb arms in the F2 and F3 hybrids were correlated with the frequencies of recombined Hv chromosomes in progenies. Self-generation of hybrids, the number of Hv and Hb chromosomes, and the number of recombined Hv chromosomes of the hybrids strongly influenced the Hv-Hb pairing frequency in meiosis. Within the offspring of F2 and F3 hybrids both Hv plants and hybrids were detected. In contrast, all progenies of the F4 hybrid were hybrids which exhibited centromere misdivisions. The highest frequencies of homoeologous pairing in hybrids and most recombinants were obtained for the barley chromosome 1HL. Recombinants for 4HL, 5HS, 6HS, and 7HS were rarely found. Meiotic pairing and recombinants involving chromosome 1HS were never observed. The results of this study demonstrate that fertile tetraploid interspecific hybrids with a high intergenomic pairing at meiosis are valuable basic material for introgression breeding in barley.

Dynamics and metastable surface structure of double atomic layer of water molecules and ions at the interface between KBr(c) and water

2004 ◽  
Vol 76 (1) ◽  
pp. 115-122 ◽  
Author(s):  
K. Ichikawa ◽  
S. Sato ◽  
N. Shimomura
Keyword(s):  
Surface Structure ◽  
Vertical Motion ◽  
Atomic Layer ◽  
Water Molecules ◽  
Potassium Ions ◽  
Force Microscopy ◽  
Structure And Dynamics ◽  
Atomic Force ◽  
Bromide Ions

The metastable surface structure and dynamics of water molecules, cations, and anions at the interface between KBr(001) and water have been demonstrated from the images in situ observed in atomic resolution using atomic force microscopy. The vertical motion of potassium ions, which means their own transfer from the equilibrium sites to the upper height right on the underlying bromide ions, has been observed at the interface. They are used to be located in some steady state stabilized by their interaction with water molecules in the double atomic layer at the interface. The observed water molecules bridge two bromide ions by hydrogen bond; the water molecules are sandwiched by the potassium ions and vice versa.

scholarly journals Estimating long-term groundwater storage and its controlling factors in Alberta, Canada

2018 ◽  
Vol 22 (12) ◽  
pp. 6241-6255 ◽  
Author(s):  
Soumendra N. Bhanja ◽  
Xiaokun Zhang ◽  
Junye Wang
Keyword(s):  
Groundwater Recharge ◽  
Environmental Sustainability ◽  
Land Surface ◽  
Water Withdrawal ◽  
Groundwater Storage ◽  
Budget Analysis ◽  
Terrestrial Water ◽  
The Individual ◽  
Gravity Recovery

Abstract. Groundwater is one of the most important natural resources for economic development and environmental sustainability. In this study, we estimated groundwater storage in 11 major river basins across Alberta, Canada, using a combination of remote sensing (Gravity Recovery and Climate Experiment, GRACE), in situ surface water data, and land surface modeling estimates (GWSAsat). We applied separate calculations for unconfined and confined aquifers, for the first time, to represent their hydrogeological differences. Storage coefficients for the individual wells were incorporated to compute the monthly in situ groundwater storage (GWSAobs). The GWSAsat values from the two satellite-based products were compared with GWSAobs estimates. The estimates of GWSAsat were in good agreement with the GWSAobs in terms of pattern and magnitude (e.g., RMSE ranged from 2 to 14 cm). While comparing GWSAsat with GWSAobs, most of the statistical analyses provide mixed responses; however the Hodrick–Prescott trend analysis clearly showed a better performance of the GRACE-mascon estimate. The results showed trends of GWSAobs depletion in 5 of the 11 basins. Our results indicate that precipitation played an important role in influencing the GWSAobs variation in 4 of the 11 basins studied. A combination of rainfall and snowmelt positively influences the GWSAobs in six basins. Water budget analysis showed an availability of comparatively lower terrestrial water in 9 of the 11 basins in the study period. Historical groundwater recharge estimates indicate a reduction of groundwater recharge in eight basins during 1960–2009. The output of this study could be used to develop sustainable water withdrawal strategies in Alberta, Canada.

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