Ultrastructural localization of adenosinetriphosphatase activity in skeletal muscle by calcium precipitation at high pH

1969 ◽  
Vol 3 (1) ◽  
pp. 307-323
Author(s):  
M. V. Vye ◽  
D. A. Fischman ◽  
J. L. Hansen
Keyword(s):  
Skeletal Muscle ◽  
Ultrastructural Localization ◽  
High Ph ◽  
Calcium Precipitation

scholarly journals Ultrastructural localization of intracellular calcium stores by a new cytochemical method.

1987 ◽  
Vol 35 (9) ◽  
pp. 939-956 ◽  
Author(s):  
M Poenie ◽  
D Epel
Keyword(s):  
Skeletal Muscle ◽  
Intracellular Calcium ◽  
Sea Urchin ◽  
Acrosome Reaction ◽  
Ultrastructural Localization ◽  
Calcium Stores ◽  
Rat Skeletal Muscle ◽  
Cytochemical Method ◽  
Calcium Localization ◽  
Sea Urchin Sperm

We describe a new cytochemical method for ultrastructural localization of intracellular calcium stores. This method uses fluoride ions for in situ precipitation of intracellular calcium during fixation. Comparisons made using oxalate, antimonate, or fluoride showed that fluoride was clearly superior for intracellular calcium localization in eggs of the sea urchin Strongylocentrotus purpuratus. Whereas oxalate generally gave no intracellular precipitate and antimonate gave copious but random precipitate, three prominent calcium stores were detected using fluoride: the tubular endoplasmic reticulum, the cortical granules, and large, clear, acidic vesicles of unknown function. The mitochondria of these eggs generally showed no detectable calcium deposits. X-ray spectra confirmed the presence of calcium in the fluoride precipitates, although in some cases magnesium was also detected. Rat skeletal muscle and sea urchin sperm were used to test the reliability of the fluoride method for calcium localization. In rat skeletal muscle, most fluoride precipitate was confined to the sarcoplasmic reticulum. Using sea urchin sperm, which transport calcium into the mitochondria after exposure to egg jelly to induce the acrosome reaction, the expected result was also obtained. Before the acrosome reaction, sperm mitochondria contain no detectable calcium-containing precipitate. Within 4 min after induction of the acrosome reaction, the expected result was also obtained. Before the acrosome reaction, sperm mitochondria displayed many foci of calcium-containing precipitate. The use of fluoride for intracellular calcium localization therefore appears to be a substantial improvement over previous cytochemical methods.

The Ultrastructural Localization of Antigens for Skeletal Muscle Antibodies in Myasthenia Gravis

1987 ◽  
Vol 505 (1 Myasthenia Gr) ◽  
pp. 732-734 ◽  
Author(s):  
PER R. FLOOD ◽  
ROGER BJUGN ◽  
NILS E. GILHUS ◽  
HÅKON HOFSTAD ◽  
ROALD MATRE ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Myasthenia Gravis ◽  
Ultrastructural Localization ◽  
Skeletal Muscle Antibodies

Ultrastructural localization of myoglobin mRNA in human skeletal muscle

1994 ◽  
Vol 101 (2) ◽  
pp. 99-104 ◽  
Author(s):  
T. Mitsui ◽  
H. Kawai ◽  
T. Naruo ◽  
S. Saito
Keyword(s):  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
Ultrastructural Localization

scholarly journals Localization of C-protein isoforms in chicken skeletal muscle: ultrastructural detection using monoclonal antibodies.

1984 ◽  
Vol 98 (4) ◽  
pp. 1514-1522 ◽  
Author(s):  
J E Dennis ◽  
T Shimizu ◽  
F C Reinach ◽  
D A Fischman
Keyword(s):  
Skeletal Muscle ◽  
Monoclonal Antibodies ◽  
Fiber Type ◽  
Ultrastructural Localization ◽  
Absolute Number ◽  
Protein Isoforms ◽  
C Protein ◽  
Thick Filaments ◽  
C Proteins ◽  
Chicken Skeletal Muscle

Monoclonal antibodies (McAbs) specific for the fast (MF-1) and slow (ALD-66) isoforms of C-protein from chicken skeletal muscle have been produced and characterized. Using these antibodies it was possible to demonstrate that skeletal muscles of varying fiber type express different isoforms of this protein and that in the posterior latissimus dorsi muscle both isoforms are co-expressed in the same myofiber (17, 18). Since we had shown that both isoforms were present in all sarcomeres, it was feasible to test whether the two isoforms co-distributed in the same 43-nm repeat within the A-band, thereby establishing a minimum number of C-proteins per repeat in the thick filaments. Here we describe the ultrastructural localization of C-protein in myofibers from three muscle types of the chicken using these same McAbs. We observed that although C-protein was present in a 43-nm repeat along the filaments in all three muscles, there were marked differences in the absolute number and position occupied by the different isoforms. Since McAbs MF-1 and ALD-66 decorated the same 43-nm repeats in the A-bands of the posterior latissimus dorsal muscle, we suggest that at least two C-proteins can co-localize at binding sites 43 nm apart along thick filaments of this muscle.

scholarly journals Ultrastructural Localization of Glycolytic Enzymes on Sarcoplasmic Reticulum Vesicles

1998 ◽  
Vol 46 (4) ◽  
pp. 419-427 ◽  
Author(s):  
Kai Y. Xu ◽  
Lewis C. Becker
Keyword(s):  
Skeletal Muscle ◽  
Sarcoplasmic Reticulum ◽  
Pyruvate Kinase ◽  
Indirect Evidence ◽  
Ultrastructural Localization ◽  
Immunogold Labeling ◽  
Glycolytic Enzymes ◽  
Electron Microscopic ◽  
Gold Particles ◽  
Cytoplasmic Side

We have previously obtained indirect evidence that sarcoplasmic reticulum (SR) vesicles from cardiac and skeletal muscle contain the complete chain of glycolytic enzymes from aldolase to pyruvate kinase. To investigate directly whether pyruvate kinase and other glycolytic enzymes are anatomically associated with the SR, electron microscopic immunogold labeling studies were carried out in isolated SR vesicles using specific primary antibodies against selected glycolytic enzymes and Ca2+-ATPase, and appropriate secondary antibodies labeled with 6-nm or 12-nm gold particles. Pyruvate kinase was broadly dispersed on the cytoplasmic side of the SR membrane of both cardiac and skeletal muscle vesicles. With 6-nm gold particles, the density of binding to pyruvate kinase was 2522 ± 445 and 4171 ± 1379 particles/μm2 for cardiac and skeletal muscle SR, respectively. Binding densities to Ca2+-ATPase were similar (2550 ± 639 particles/μm2 for cardiac SR, 3877 ± 408 particles/μm2 for skeletal muscle SR). Immunogold labeling of ultrathin sections indicated that pyruvate kinase was attached to the SR membrane and located immediately adjacent to the Ca2+-ATPase. Aldolase and glyceraldehyde phosphate dehydrogenase were also found to be attached to the cytoplasmic side of SR vesicles and located in close proximity to Ca2+-ATPase. These results provide the first ultrastructural evidence that glycolytic enzymes are anatomically associated with SR membranes and located near the SR Ca2+-ATPase. The results further support the hypothesis that ATP generated by SR-associated glycolytic enzymes is coupled to SR Ca2+ active transport.

scholarly journals Ultrastructural localization of acetylcholinesterase in cultured cells. I. Embryo muscle.

1976 ◽  
Vol 24 (9) ◽  
pp. 969-978 ◽  
Author(s):  
H R Sawyer ◽  
T K Golder ◽  
P S Nieberg ◽  
B W Wilson
Keyword(s):  
Skeletal Muscle ◽  
Enzyme Activity ◽  
Sarcoplasmic Reticulum ◽  
Kinetic Data ◽  
Cultured Cells ◽  
Ultrastructural Localization ◽  
Potassium Ferricyanide ◽  
The Other ◽  
Staining Procedure ◽  
Staining Methods

Several techniques were employed to examine the localization of acetylcholinesterase (EC 3.1.1.7, AChE) in cultured chick embryonic skeletal muscle. Glutaraldehyde produced the best cellular preservation but less enzyme activity was lost when the cells were fixed in paraformaldehyde. Two staining methods were examined: in one (Karnovsky MJ, Roots L: J Histochem Cytochem 12:219, 1964) potassium ferricyanide was added with the primary reactants, and in the other (Tsuji S: Histochemistry 42:99, 1974) the potassium ferricyanide was added at the end of the staining procedure. Localizations of AChE were similar with both stains; activity was present in the nuclear envelope, the perinuclear sarcoplasm, the sarcoplasmic reticulum, subsurface vesicles and bound outside the cells. /owever, a granular artifact was found with the method of Karnovsky and Roots that did not appear with the method of Tsuji. The localization of AChE are consistent with kinetic data that AchE binds, moves and is released from cultured muscle fibers.

scholarly journals Ultrastructural Localization of Histidine-rich Glycoprotein in Skeletal Muscle Fibers: Colocalization With AMP Deaminase

2019 ◽  
Vol 68 (2) ◽  
pp. 139-148
Author(s):  
Letizia Mattii ◽  
Francesco Bianchi ◽  
Alessandra Falleni ◽  
Sabina Frascarelli ◽  
Matilde Masini ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Sarcoplasmic Reticulum ◽  
Laser Scanning ◽  
Muscle Fibers ◽  
Ultrastructural Localization ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Amp Deaminase ◽  
Skeletal Muscle Fibers ◽  
Nuclear Levels

Histidine-rich glycoprotein (HRG) is a plasma protein synthesized by the liver. We have given the first evidence of a tissue localization of HRG demonstrating its presence in skeletal muscle, associated with the zinc enzyme AMP deaminase (AMPD1). Moreover, we have shown that muscle cells do not synthesize HRG, but they can internalize it from plasma. We have recently demonstrated by confocal laser scanning microscopy that in human skeletal muscle, HRG is mainly localized in the myofibrils, preferentially at the I-band of the sarcomere, in the sarcoplasm, and in the nuclei. Using transmission electron microscopy and immunogold analysis, we carried out this study on human and rat normal skeletal muscles with the purpose to deepen the ultrastructural localization of HRG in skeletal muscle fibers. The immunogold analysis evidenced the presence of HRG in the sarcomeres, mainly in the I-band and to a less extent in the A-band, in the heterochromatin of nuclei, and in the sarcoplasmic reticulum. The colocalization of HRG and skeletal muscle AMPD1 was also analyzed. A colabeling of HRG and AMPD1 was evident at sarcomeric, sarcoplasmic reticulum, and nuclear levels. The significance of these interesting and new results is discussed in this article.

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