Structural Aspects of Protein Accumulation in Developing Pea Cotyledons. II. Three-Dimensional Reconstructions of Vacuoles and Protein Bodies From Serial Sections

1980 ◽  
Vol 7 (3) ◽  
pp. 329 ◽  
Author(s):  
S Craig ◽  
DJ Goodchild ◽  
C Miller
Keyword(s):  
Three Dimensional ◽  
Protein Body ◽  
Protein Bodies ◽  
Dimensional Structure ◽  
Protein Accumulation ◽  
Serial Sections ◽  
Seed Maturity ◽  
Spherical Structures ◽  
A Cell ◽  
Structural Aspects

The three-dimensional structure of vacuoles and protein bodies seen in developing cotyledons from pea (Pisum sativum L.) have been reconstructed from serial sections. At days 12 and 15 after flowering, serial sections 1 �m thick of epoxy-embedded seed tissue were used to determine vacuole morphology while, at day 20, serial sections 0.25 �m thick were examined by electron microscopy to ascertain protein body morphology. At day 12 there are one or two large vacuoles having extremely complex protrusions emanating from a larger central vacuolar volume. This gives rise to up to 20 apparently discrete vacuole profiles in a given section through a cell. By day 15, there are many smaller, approximately spherical, vacuoles and also some that are more complex. At day 20 most protein bodies are discrete, spherical structures, although a few irregularly shaped bodies are seen. The results support the concept of a large highly convoluted central vacuole fragmenting to give rise to the protein bodies seen towards seed maturity.

Structural Aspects of Protein Accumulation in Developing Pea Cotyledons. III. Immunocytochemical Localization of Legumin and Vicilin Using Antibodies Shown to Be Specific by the Enzyme-Linked Immunosorbent Assay (ELISA)

1980 ◽  
Vol 7 (3) ◽  
pp. 339 ◽  
Author(s):  
S Craig ◽  
A Millerd ◽  
DJ Goodchild
Keyword(s):  
Immunosorbent Assay ◽  
Storage Proteins ◽  
Protein Body ◽  
Protein Bodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Accumulation ◽  
Immunocytochemical Localization ◽  
Elisa Technique ◽  
Immunocytochemical Techniques ◽  
Structural Aspects

The site of sequestration of the storage proteins legumin and vicilin during development of cotyledons from pea (Pisum sativum L.) has been determined using improved immunocytochemical techniques. Antibodies to legumin and vicilin were made monospecific by affinity chromatography. They were allowed to react on sections of glycol methacrylate-embedded cotyledon tissue and detected by indirect immunocytochemical localization using rhodamine-labelled antibodies. The enzyme-linked immunosorbent assay (ELISA) technique was adapted to verify antibody specificity at a sensitivity up to 300 times greater than that of immunodiffusion. Legumin and vicilin 4 are localized in small peripheral deposits within large vacuoles as early as day 8 after flowering. As the vacuoles fragment during development the storage proteins continue to be localized in the vacuolar deposits until, at day 16, they entirely fill vacuoles, now termed protein bodies. Thereafter, the protein bodies become more densely packed and retain a similar form from day 22 to maturity. Wherever the same vacuolar deposit of protein body could be observed in adjacent sections, antilegumin and antivicilin 4 labelled both deposits, clearly indicating that both storage proteins are sequestered into the same area of protein.

Three-Dimensional Reconstruction Using Stereo Pairs from Serial Thick Sections

1977 ◽  
Vol 35 ◽  
pp. 562-563
Author(s):  
Robert Glaeser ◽  
Thomas Bauer ◽  
David Grano
Keyword(s):  
Three Dimensional ◽  
Dimensional Structure ◽  
Serial Sections ◽  
Thin Sections ◽  
3 Dimensional ◽  
Transmission Electron ◽  
Freeze Dried ◽  
Dimensional Reconstruction ◽  
Stereo Imagery ◽  
Whole Mounts

In transmission electron microscopy, the 3-dimensional structure of an object is usually obtained in one of two ways. For objects which can be included in one specimen, as for example with elements included in freeze- dried whole mounts and examined with a high voltage microscope, stereo pairs can be obtained which exhibit the 3-D structure of the element. For objects which can not be included in one specimen, the 3-D shape is obtained by reconstruction from serial sections. However, without stereo imagery, only detail which remains constant within the thickness of the section can be used in the reconstruction; consequently, the choice is between a low resolution reconstruction using a few thick sections and a better resolution reconstruction using many thin sections, generally a tedious chore. This paper describes an approach to 3-D reconstruction which uses stereo images of serial thick sections to reconstruct an object including detail which changes within the depth of an individual thick section.

scholarly journals Microtubular apparatus of melanophores: three dimensional organization

1978 ◽  
Vol 76 (3) ◽  
pp. 605-614 ◽  
Author(s):  
M Schliwa
Keyword(s):  
Three Dimensional ◽  
Cell Process ◽  
Serial Sections ◽  
Dimensional Model ◽  
Three Dimensional Model ◽  
Cell Region ◽  
Thin Sectioning ◽  
A Cell ◽  
Central Apparatus ◽  
Pterophyllum Scalare

Microtubular organization in the melanophores of the angelfish, Pterophyllum scalare, has been studied by serial thin sectioning. The course of microtubules has been followed in sets of transverse serial sections taken from the centrosphere and a segment of a cell process, respectively. Microtubules arise from a prominent zone in the cell center, the central apparatus, which is composed of numerous, small, electron-dense aggregates. the number of these loosely distributed densities is highest in the center of the centrosphere, but they may also be found at its periphery. Microtubules insert into, or becomes part of, the dense material, or at least start in its vicinity. Dense aggregates may be separated from centrioles by several micrometers rather than only being closely associated with these organelles. At some distance from the organizing zone, most of the microtubules gradually assume a cortical arrangement, i.e., take a course within about 100 nm of the limiting membrane. Serial sections were used to trace all microtubules within a 6μm-long segment of a cell process. 94 percent of the microtubules observed in this segment run its entire length; it is conceivable, therefore that a considerable number of microtubules extend between the initiating site in the centrosphere and the outermost cell region. A three-dimensional model of the 6μm-long segment reveals that, despite changes in the cell process outline, microtubules maintain a strictly cortical arrangement which gives the impression of a microtubule "palisade" lining the cortex of the cell process. The features of the microtubular apparatus of angelfish melanophores are discussed in relation to factors controlling microtubule initiation and distribution.

The Centrosome: Three-Dimensional Structure of a Cell Organelle at 40A Resolution Obtained by Electrontomography

1998 ◽  
Vol 4 (S2) ◽  
pp. 436-437
Author(s):  
T. Ruiz ◽  
M. Radermacher ◽  
B. Rath ◽  
C. Rieder ◽  
M. Bornens
Keyword(s):  
Three Dimensional ◽  
Dimensional Structure ◽  
Cell Organelle ◽  
Microtubule Network ◽  
Detailed Understanding ◽  
Alignment Procedure ◽  
Carbon Coated ◽  
Pericentriolar Material ◽  
A Cell ◽  
Filamentous Material

The organization of the microtubule network of eukariotic cells is controlled by the centrosome. This organelle which duplicates once during the cell cycle comprises two centrioles and a dense filamentous material called pericentriolar material. The centrioles are cylinders of ≈ 0.25 μm in diameter and ≈ 0.5μm in length and are composed of 9 microtubule triplets at the proximal end and 9 doublets at the distal end. For the last 50 years the centrosome and the centrioles in particular have been the object of many optical and electron microscopy studies. But, in spite of their highly symmetric organization, their fine structure is still not well understood. The reconstruction presented here is a substantial step forward towards a more detailed understanding of their structure.Centrosomes were isolated from a human lymphoblastoma cell line (1). 10 nm gold particles were applied to formvar-carbon coated grids to provide markers for the alignment procedure (2,3). Centrosomes were centrifuged onto the grids and negatively stained.

Three dimensional structure of the leishmania amastigote as revealed by computer-aided reconstruction from serial sections

Parasitology ◽  
1986 ◽  
Vol 92 (1) ◽  
pp. 13-23 ◽  
Author(s):  
G. H. Coombs ◽  
L. Tetley ◽  
V. A. Moss ◽  
K. Vickerman
Keyword(s):  
Cell Volume ◽  
Three Dimensional ◽  
Spatial Arrangement ◽  
Dimensional Structure ◽  
Serial Sections ◽  
Leishmania Mexicana ◽  
Cell Organelles ◽  
3 Dimensional ◽  
Computer Aided ◽  
Total Cell Volume

SUMMARYComputer-aided reconstruction from serial sections has been used to analyse the 3-dimensional structure of entire amastigotes of Leishmania mexicana mexicana and to determine the number, arrangement and volume of each organelle. In two reconstructions, the lysosome-like ‘megasomes’ were the most numerous organelle, there being 34 in one amastigote, and they comprised as much as 15% of the total cell volume. In contrast, as few as 9 glycosomes were present, accounting for less than 1% of the cell volume. The unitary nature of the mitochondrion was confirmed and its complex basket-like structure was revealed. The spatial arrangement of the cell organelles is here displayed in stereo-pairs.

Three-dimensional structure of a cell wall glycoprotein

1982 ◽  
Vol 162 (2) ◽  
pp. 459-471 ◽  
Author(s):  
P.J. Shaw ◽  
G.J. Hills
Keyword(s):  
Cell Wall ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Three Dimensional Structure ◽  
A Cell

Three-dimensional structural aspects of the design of new protein molecules

1986 ◽  
Vol 317 (1540) ◽  
pp. 333-344 ◽  
Keyword(s):  
Tertiary Structure ◽  
Protein Structures ◽  
Three Dimensional ◽  
Spare Parts ◽  
Dimensional Structure ◽  
Sequence Patterns ◽  
Essential Prerequisite ◽  
Structure Of Proteins ◽  
New Protein ◽  
Structural Aspects

A knowledge of the three-dimensional structure of proteins is an essential prerequisite for the design of new molecules. When the tertiary structure is not available from high-resolution X-ray or n.m.r. analysis, the success of prediction is improved by using a relational database of known protein structures. This can be searched to provide information on secondary structure motifs and domains which are recognized by characteristic sequence patterns and which are assembled as ‘spare parts’ by using computer graphics. Similar techniques can be used to give approximate structures for amino-acid replacements, deletions and insertions introduced by mutagenesis. The resulting structures are optimized by using interactive graphics, energy minimization and molecular dynamics.

scholarly journals SERIAL RECONSTRUCTION OF THE CHARACTERISTIC GRANULE OF THE LANGERHANS CELL

1968 ◽  
Vol 36 (3) ◽  
pp. 595-602 ◽  
Author(s):  
Richard W. Sagebiel ◽  
Thomas H. Reed
Keyword(s):  
Electron Microscope ◽  
Electron Micrographs ◽  
Three Dimensional ◽  
Langerhans Cell ◽  
Human Epidermis ◽  
Dimensional Structure ◽  
Serial Sections ◽  
Three Dimensional Structure ◽  
Three Dimensional Models ◽  
The Many

Three-dimensional models of individual granules in the same Langerhans cell were made after analyzing serial sections of human epidermis in the electron microscope. These models revealed that the granule is made up of a flattened or curved orthogonal net of particles which is bounded externally by a limiting membrane and which may be disc-shaped, cup-shaped, or combinations of both shapes. This variety of shapes accounts for the many configurations of the granule seen in individual electron micrographs. Usually, the granule has a vesicular portion at, or near one margin. This demonstration of the three-dimensional structure of the granule establishes the inaccuracy of previously used descriptive terms, the granule should be called simply the "Langerhans cell granule."

Structural Aspects of Protein Accumulation in Developing Pea Cotyledons. I. Qualitative and Quantitative Changes in Parenchyma Cell Vacuoles

1979 ◽  
Vol 6 (1) ◽  
pp. 81 ◽  
Author(s):  
S Craig ◽  
DJ Goodchild ◽  
AR Hardham
Keyword(s):  
Cell Volume ◽  
Surface Area ◽  
Structural Changes ◽  
Light And Electron Microscopy ◽  
Parenchyma Cell ◽  
Protein Body ◽  
Average Diameter ◽  
Protein Bodies ◽  
Protein Accumulation ◽  
Parenchyma Cells

Structural changes in pea cotyledons during development were studied using light and electron microscopy. Changes in the vacuolar system and cytoplasm of cotyledon parenchyma cells, during the period of storage protein deposition, are reported. Eight days after flowering, the parenchyma cells each contain one or two large vacuoles that are replaced by progressively smaller vacuoles during the next 10 days of development. Stainable material that can be histochemically identified as protein appears on the inner surface of the vacuole tonoplast 8 days after flowering. These vacuoles become smaller and more frequent during development and the amount of proteinaceous material within each vacuole increases until, at days 16-20 after flowering, they become densely packed with protein and are described as protein bodies. At day 8, the vacuole(s) have an average diameter of 39 �m, an average volume of 41 000 �m� , representing 75 % of the cell volume, and a surface area of 5500 �m�. By day 20, the average protein body diameter has fallen to 1 �m. There are, however, approx. 175 000 such protein bodies per cell, occupying 91 500 �m� or approx. 20 % of the cell volume, and whose total surface area is 550 000 �m�. The surface to volume ratlo of the vacuole/protein bodies Increases 55 times between days 8 and 20. Apart from this increase in surface area available for possible entry of protein, no mechanism for such entry can be suggested from our nlicrographs.

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