Electron microscopic tomography of rat-liver mitochondria and their interactions with the endoplasmic reticulum

BioFactors ◽  
1998 ◽  
Vol 8 (3-4) ◽  
pp. 225-228 ◽  
Author(s):  
Carmen A. Mannella ◽  
Karolyn Buttle ◽  
Bimal K. Rath ◽  
M. Marko
Keyword(s):  
Endoplasmic Reticulum ◽  
Rat Liver ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Electron Microscopic

scholarly journals Electron microscopic localization of glutamate dehydrogenase in rat liver mitochondria by an immunogold procedure and monoclonal and polyclonal antibodies.

1986 ◽  
Vol 34 (7) ◽  
pp. 913-922 ◽  
Author(s):  
E Knecht ◽  
A Martinez-Ramon ◽  
S Grisolia
Keyword(s):  
Glutamate Dehydrogenase ◽  
Rat Liver ◽  
Polyclonal Antibodies ◽  
Protein A ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Electron Microscopic ◽  
Cross Sectional ◽  
Gold Particles ◽  
Parenchymal Cells

Glutamate dehydrogenase (GDH) was localized in rat liver by indirect electron microscopic immunogold, using different sizes of gold particles and monoclonal and polyclonal antibodies. Using the protein A-gold technique in double immunocytochemical experiments, both antibodies, at their optimal dilutions, gave similar results. A novel assessment of the distribution of GDH was made by measurements of the number of gold particles per square micrometer of cross-sectional images of individual mitochondria. The data indicate intracellular homogeneity among mitochondria in individual parenchymal cells. The enzyme is almost absent in non-parenchymal cells. Finally, GDH was found mainly in association with the mitochondrial inner membrane.

scholarly journals EFFECT OF ACTIVE ACCUMULATION OF CALCIUM AND PHOSPHATE IONS ON THE STRUCTURE OF RAT LIVER MITOCHONDRIA

1964 ◽  
Vol 23 (1) ◽  
pp. 21-38 ◽  
Author(s):  
John W. Greenawalt ◽  
Carlo S. Rossi ◽  
Albert L. Lehninger
Keyword(s):  
Specific Gravity ◽  
Rat Liver ◽  
Cesium Chloride ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Electron Microscopic ◽  
Dense Granules ◽  
Phosphate Ions ◽  
Microscopic Studies

Rat liver mitochondria allowed to accumulate maximal amounts of Ca++ and HPO4= ions from the suspending medium in vitro during respiration have a considerably higher specific gravity than normal mitochondria and may be easily separated from the latter by isopycnic centrifugation in density gradients of sucrose or cesium chloride. When the mitochondria are allowed to accumulate less than maximal amounts of Ca++ and HPO4= from the medium, they have intermediate specific gravities which are roughly proportional to their content of calcium phosphate. Maximally "loaded" mitochondria are relatively homogeneous with respect to specific gravity. Correlated biochemical and electron microscopic studies show that Ca++-loaded mitochondria contain numerous dense granules, of which some 85 per cent are over 500 A in diameter. These granules are electron-opaque not only following fixation and staining with heavy metal reagents, but also following fixation with formaldehyde, demonstrating that the characteristic granules in Ca++-loaded mitochondria have intrinsic electron-opacity. The dense granules are almost always located within the inner compartment of the mitochondria and not in the space between the inner and outer membranes. They are frequently located at or near the cristae and they often show electron-transparent "cores." Such granules appear to be made up of clusters of smaller dense particles, but preliminary x-ray diffraction analysis and electron diffraction studies have revealed no evidence of crystallinity in the deposits. The electron-opaque granules decrease in number when the Ca++-loaded mitochondria are incubated with 2,4-dinitrophenol; simultaneously there is discharge of Ca++ and phosphate from the mitochondria into the medium.

Electron Microscopic Tomography of Whole, Frozen-Hydrated Rat-Liver Mitochondria at 400 kv

1999 ◽  
Vol 5 (S2) ◽  
pp. 416-417
Author(s):  
C.A. Mannella ◽  
C.-E Hsieh ◽  
M. Marko
Keyword(s):  
Rat Liver ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Peripheral Compartment ◽  
Collection System ◽  
Electron Microscopic ◽  
Automated Data Collection ◽  
Outer Membranes ◽  
Mitochondrial Inner Membrane ◽  
Structural Preservation

Electron microscopic tomography is providing important new insights about the internal structure of the mitochondrion. In particular, the infoldings of the mitochondrial inner membrane (cristae), which are usually rendered as lamelliform baffles, are revealed to have considerable tubular nature. Rather than opening wide to the peripheral compartment (between the inner and outer membranes), the cristae connect to the outside and to each other through narrow (20-30 nm) tubular segments, which can be hundreds of nanometers long. This suggests that diffusion of ions, metabolites and proteins between the intracristal and intermembrane spaces may be restricted.The earlier tomographic reconstructions were done on conventionally prepared, plastic-embedded specimens, which raises the usual concerns about structural preservation. More recently, we have undertaken tomography of isolated rat-liver mitochondria that have been embedded in vitreous ice (by plunge-freezing in iso-osmotic buffer without chemical fixatives or stains). These frozen hydrated specimens are imaged with a JEOL 4000FX equipped with a Gatan cryo-transfer holder and a Tietz automated data collection system, with a Ik × Ik CCD. For 3D reconstructions, images were recorded at a dose of 5 e−Å2 at 2° increments over the range +/− 60°.

Electron Microscopic Tomography of Cellular Organelles: Chemical Fixation Vs. Cryo-Substitution of Rat-Liver Mitochondria

1998 ◽  
Vol 4 (S2) ◽  
pp. 430-431
Author(s):  
C.A. Mannella ◽  
K. Buttle ◽  
K. Tessitore ◽  
B.K. Rath ◽  
C. Hsieh ◽  
...  
Keyword(s):  
Rat Liver ◽  
3D Structure ◽  
Design Feature ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Electron Microscopic ◽  
Inner Membrane ◽  
Low Contrast ◽  
Mitochondrial Inner Membrane ◽  
Cellular Organelles

Electron microscopic tomography is proving to be a valuable tool for investigating the 3D structure and organization of cellular organelles. Important progress is being made in the application of the technique to frozen-hydrated material, but it is likely that success with thick specimens will be limited by the low contrast and beam sensitivity of naked biological material. Thus, optimizing procedures for fixing, embedding, staining, and selectively labelling cells for 3D electron microscopy remains a priority.Tomography of chemically fixed and plastic-embedded rat-liver tissue and isolated mitochondria has shown that the cristae (the invaginations of the mitochondrial inner membrane) are pleiomorphic and connected to each other and to the surface of the inner membrane by tubular regions 30-40 nm in diameter. This basic design feature has important implications for the microcompartmentation of ions and molecules within this organelle.

scholarly journals SEDIMENTATION PROPERTIES OF RAT LIVER MITOCHONDRIA

1970 ◽  
Vol 46 (1) ◽  
pp. 17-26 ◽  
Author(s):  
John N. Loeb ◽  
Daniel V. Kimberg
Keyword(s):  
Rat Liver ◽  
Gradient Centrifugation ◽  
Sucrose Density Gradient ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Electron Microscopic ◽  
Sucrose Density Gradient Centrifugation ◽  
Sedimentation Behavior ◽  
Sucrose Medium ◽  
Mitochondrial Volume

A prediction of the velocity of sedimentation of rat liver mitochondria in sucrose gradients is made on the basis of recent measurements of the size of isolated mitochondria suspended in sucrose medium and the model proposed by Bentzel and Solomon to describe the osmotic behavior of mitochondria. The experimentally observed velocity is extremely close to the predicted value and confirms by a different approach the estimate of mitochondrial volume made by Baudhuin and Berthet on the basis of electron microscopic measurements. Because cortisone treatment of rats is known to result in a marked increase in mitochondrial size as observed under the electron microscope, mitochondria were co-isolated from livers of control and cortisone-treated animals, and the sedimentation behavior of the mixtures was examined by sucrose density gradient centrifugation. Mitochondria from cortisone-treated animals were found to sediment 1.4 times as rapidly as those from control animals, indicating that their increased size cannot entirely be due to an increased imbibition of fluid from the surrounding sucrose medium, and that the change in size must at least in part be due to a change in content of nondiffusible mitochondrial components. Although the increase in sedimentation velocity of mitochondria from cortisone-treated animals is striking, it is less than that predicted solely on the basis of their size relative to that of control mitochondria. It is concluded that the increases in mitochondrial size and content of nondiffusible components produced by cortisone treatment are accompanied by alterations in mitochondrial composition as well.

Sign in / Sign up

Export Citation Format

Share Document