scholarly journals Ultrastructural localization of the Ca2+ + Mg2+-dependent ATPase of sarcoplasmic reticulum in rat skeletal muscle by immunoferritin labeling of ultrathin frozen sections.

1982 ◽  
Vol 92 (2) ◽  
pp. 409-416 ◽  
Author(s):  
A O Jorgensen ◽  
A C Shen ◽  
D H MacLennan ◽  
K T Tokuyasu
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Plasma Membranes ◽  
Ultrastructural Localization ◽  
Published Data ◽  
Frozen Sections ◽  
Dependent Atpase ◽  
Sarcoplasmic Reticulum Membrane ◽  
Ultrathin Frozen Sections ◽  
Terminal Cisternae ◽  
Simultaneous Visualization

The ultrastructural localization of the Ca2+ + Mg2+-dependent ATPase of sarcoplasmic reticulum in rat gracilis muscle was determined by indirect immunoferritin labeling of ultrathin frozen sections. Simultaneous visualization of ferritin particles and of adsorption-stained cellular membranes showed that the Ca2+ + Mg2+-ATPase was concentrated in the longitudinal sarcoplasmic reticulum and in the nonjunctional regions of the terminal cisternae membrane but was virtually absent from mitochondria, plasma membranes, transverse tubules, and junctional sarcoplasmic reticulum. Ferritin particles were found preponderantly on the cytoplasmic surface of the membrane, in agreement with published data showing an asymmetry of the Ca2+ + Mg2+-ATPase within the sarcoplasmic reticulum membrane. Comparison of the density of ferritin particles in fast and slow myofibers suggested that the density of the Ca2+ + Mg2+-ATPase in the sarcoplasmic reticulum membrane in a fast myofiber is approximately two times higher than in a slow myofiber.

scholarly journals Ultrastructural localization of calsequestrin in adult rat atrial and ventricular muscle cells.

1985 ◽  
Vol 101 (1) ◽  
pp. 257-268 ◽  
Author(s):  
A O Jorgensen ◽  
A C Shen ◽  
K P Campbell
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Ultrastructural Localization ◽  
Major Site ◽  
Cardiac Sarcoplasmic Reticulum ◽  
Adult Rat ◽  
Dependent Atpase ◽  
Sarcoplasmic Reticulum Membrane ◽  
Structural Differences ◽  
Ultrathin Frozen Sections ◽  
Junctional Sarcoplasmic Reticulum

The distribution of calsequestrin in rat atrial and ventricular myocardial cells was determined by indirect immunocolloidal gold labeling of ultrathin frozen sections. The results presented show that calsequestrin is confined to the sarcoplasmic reticulum where it is localized in the lumen of the peripheral and the interior junctional sarcoplasmic reticulum as well as in the lumen of the corbular sarcoplasmic reticulum, but absent from the lumen of the network sarcoplasmic reticulum. Comparison of these results with our previous studies on the distribution of the Ca2+ + Mg2+-dependent ATPase of the cardiac sarcoplasmic reticulum show directly that the Ca2+ + Mg2+-dependent ATPase and calsequestrin are confined to distinct regions within the continuous sarcoplasmic reticulum membrane. Assuming that calsequestrin provides the major site of Ca2+ sequestration in the lumen of the sarcoplasmic reticulum, the results presented support the idea that both junctional (interior and peripheral) and specialized nonjunctional (corbular) regions of the sarcoplasmic reticulum are involved in Ca2+ storage and possibly release. Furthermore, the structural differences between the junctional and the corbular sarcoplasmic reticulum support the possibility that Ca2+ storage and/or release from the lumen of the junctional and the corbular sarcoplasmic reticulum are regulated by different physiological signals.

scholarly journals Ultrastructural localization of calsequestrin in rat skeletal muscle by immunoferritin labeling of ultrathin frozen sections.

1983 ◽  
Vol 97 (5) ◽  
pp. 1573-1581 ◽  
Author(s):  
A O Jorgensen ◽  
A C Shen ◽  
K P Campbell ◽  
D H MacLennan
Keyword(s):  
Skeletal Muscle ◽  
Sarcoplasmic Reticulum ◽  
Central Region ◽  
Ultrastructural Localization ◽  
Frozen Sections ◽  
Rat Skeletal Muscle ◽  
Major Site ◽  
Ultrathin Frozen Sections ◽  
Terminal Cisternae

The ultrastructural localization of calsequestrin in rat skeletal muscle (gracilis) was determined by indirect immunoferritin labeling of ultrathin frozen sections. Calsequestrin was found in the lumen of transversely and longitudinally oriented terminal cisternae but was absent from most of the longitudinal sarcotubules and the fenestrated sarcoplasmic reticulum. Calsequestrin was occasionally observed in vesicular structures found in the central region of the I band. Since calsequestrin is believed to provide the major site of Ca2+ sequestration in the sarcoplasmic reticulum, the present results support the view that Ca2+, transported to the lumen of the sarcoplasmic reticulum, is preferentially sequestered in the terminal cisternae, but they also suggest that additional Ca2+ sequestration may occur near the center of the I band.

scholarly journals ULTRASTRUCTURAL LOCALIZATION OF INTRACELLULAR IMMUNE GLOBULINS IN PLASMA CELLS AND LYMPHOBLASTS BY ENZYME-LABELED ANTIBODIES

1969 ◽  
Vol 17 (4) ◽  
pp. 211-224 ◽  
Author(s):  
ELIZABETH H. LEDUC ◽  
GEOFFREY B. SCOTT ◽  
STRATIS AVRAMEAS
Keyword(s):  
Immune Response ◽  
Plasma Cells ◽  
Ultrastructural Localization ◽  
Frozen Sections ◽  
Cytoplasmic Staining ◽  
Specific Antibodies ◽  
Perinuclear Space ◽  
Cacodylate Buffer ◽  
Immune Globulins ◽  
Ultrathin Frozen Sections

Techniques for localization of immune globulins by specific antibodies labeled with peroxidase or alkaline phosphatase were applied to spleens of rabbits hyperimmunized against various antigens. Best results were obtained with tissue blocks or thick sections after fixation in 1% formaldehyde (freshly prepared from paraformaldehyde) in cacodylate buffer with sucrose. The edges of blocks gave adequate penetration, comparable to thick sections. Ultrathin frozen sections which were reacted directly with labeled conjugate were less successful and glycol methacrylate sections failed to stain. Results were 3-fold: (1) immune globulins were demonstrated in ergastoplasmic cisternae, perinuclear space and Golgi apparatus of plasmocytes; (2) associated ribosomal staining varied somewhat with fixation and osmolarity of fixative; (3) lymphoblasts were shown to contain antibody. Generalized cytoplasmic staining was observed in the lymphoblasts, with no localization in the ergastoplasm and with no staining of control preparations. The significance of ribosomal and lymphoblast staining is discussed in relation to cell differentiation and the development of the immune response.

scholarly journals The effect of dantrolene on skeletal-muscle sarcoplasmic-reticulum function in malignant hyperpyrexia in pigs

1983 ◽  
Vol 212 (2) ◽  
pp. 399-405 ◽  
Author(s):  
M D White ◽  
J G Collins ◽  
M A Denborough
Keyword(s):  
Skeletal Muscle ◽  
Sarcoplasmic Reticulum ◽  
Atpase Activity ◽  
Muscle Relaxant ◽  
Sodium Dodecyl Sulphate ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Malignant Hyperpyrexia ◽  
Dependent Atpase ◽  
Sarcoplasmic Reticulum Membrane

The effect of the muscle relaxant dantrolene on isolated sarcoplasmic reticulum was studied in control and malignant-hyperpyrexia-susceptible Landrace pigs. The membranes prepared from both sources showed similar Ca2+-dependent ATPase activities, had comparable phospholipid/protein ratios, and their sodium dodecyl sulphate/polyacrylamide-gel patterns were indistinguishable. Membranes from both sources appeared to bind similar amounts of dantrolene. The drug did not stimulate Ca2+-dependent ATPase activity in preparations from either source. The rates of Ca2+ exchange and Ca2+ efflux appeared to be similar in sarcoplasmic reticulum of control and malignant-hyperpyrexia-susceptible pigs. Dantrolene did not affect either the rates or the amount of Ca2+ lost from the vesicles. These results suggest that dantrolene does not directly affect the movement of Ca2+ across the sarcoplasmic-reticulum membrane.

scholarly journals NUCLEOSIDE PHOSPHATASE ACTIVITIES IN RAT CARDIAC MUSCLE

1965 ◽  
Vol 25 (2) ◽  
pp. 201-215 ◽  
Author(s):  
Edward Essner ◽  
Alex B. Novikoff ◽  
Nelson Quintana
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Cardiac Muscle ◽  
Capillary Endothelium ◽  
Frozen Sections ◽  
Phosphatase Activities ◽  
Intercalated Discs ◽  
Capillary Endothelial Cells ◽  
Contraction And Relaxation ◽  
Terminal Cisternae ◽  
T System

Localizations of aldehyde-resistant nucleoside phosphatase activities in frozen sections of rat cardiac muscle have been studied by electron microscopy. Activities are higher after fixation with formaldehyde than with glutaraldehyde. After incubation with adenosine triphosphate or inosine diphosphate at pH 7.2, reaction product is found in the "terminal cisternae" or "transverse sacs" of the sarcoplasmic reticulum, which, together with the "intermediary vesicles" (T system), constitute the "dyads" or "triads". Reaction product is also present at the membranes of micropinocytotic vacuoles which apparently form from the plasma membrane of capillary endothelial cells and from the sarcolemma. In certain regions of the intercalated discs, reaction product is found within the narrow spaces between sarcolemmas of adjacent cells and within micropinocytotic vacuoles that seem to form from the sarcolemma. With inosine diphosphate, reaction product is also found in other parts of the sarcoplasmic reticulum. After incubation with cytidine monophosphate at pH 5, reaction product is present in the transverse sacs of sarcoplasmic reticulum, in micropinocytotic vacuoles in capillary endothelium, and in lysosomes of muscle fibers and capillaries. The possible significance of the sarcoplasmic reticulum phosphatases is discussed in relation to the role the reticulum probably plays in moving calcium ions and thereby controlling contraction and relaxation of the muscle fiber.

Immunoferritin localization of intracellular antigens in positively stained frozen sections

1978 ◽  
Vol 36 (2) ◽  
pp. 168-169
Author(s):  
K. T. Tokuyasu
Keyword(s):  
Surface Tension ◽  
Water Surface ◽  
Thin Layers ◽  
The Other ◽  
Positive Staining ◽  
Frozen Sections ◽  
The Past ◽  
Ultrathin Frozen Sections ◽  
Definition Of

During the past investigations of immunoferritin localization of intracellular antigens in ultrathin frozen sections, we found that the degree of negative staining required to delineate u1trastructural details was often too dense for the recognition of ferritin particles. The quality of positive staining of ultrathin frozen sections, on the other hand, has generally been far inferior to that attainable in conventional plastic embedded sections, particularly in the definition of membranes. As we discussed before, a main cause of this difficulty seemed to be the vulnerability of frozen sections to the damaging effects of air-water surface tension at the time of drying of the sections.Indeed, we found that the quality of positive staining is greatly improved when positively stained frozen sections are protected against the effects of surface tension by embedding them in thin layers of mechanically stable materials at the time of drying (unpublished).

Localization of pancreatic enzymes in ultrathin frozen sections stained with metal labeled antibody

1978 ◽  
Vol 36 (2) ◽  
pp. 90-91
Author(s):  
Kenjiro Yasuda
Keyword(s):  
Fluorescent Antibody ◽  
Pancreatic Enzymes ◽  
Tissue Preparation ◽  
Zymogen Granules ◽  
Frozen Sections ◽  
En Bloc ◽  
Antibody Method ◽  
Ultrathin Frozen Sections ◽  
Fluorescent Antibody Method

Localization of amylase,chymotrypsinogen and trypsinogen in pancreas was demonstrated by Yasuda and Coons (1966), by using fluorescent antibody method. These enzymes were naturally found in the zymogen granules. Among them, amylase showed a diffuse localization around the nucleus, in addition to the zymogen granules. Using ferritin antibody method, scattered ferritin granules were also found around the Golgi area (Yasuda et al.,1967). The recent advance in the tissue preparation enables the antigen to be localized in the ultrathin frozen sections, by applying the labeled antibodies onto the sections instead of staining the tissue en bloc.The present study deals with the comparison of the localization of amylase and lipase demonstrated by applying the bismuth-labeled, peroxidase-labeled and ferritin-labeled antibody methods on the ultrathin frozen sections of pancreas, and on the blocks of the same tissue.

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