Quantitative Three-Dimensional Reconstructions of Excitatory Synaptic Boutons in the “Barrel Field” of the Adult “Reeler” Mouse Somatosensory Neocortex: A Comparative Fine-Scale Electron Microscopic Analysis with the Wild Type Mouse

2019 ◽  
Vol 30 (5) ◽  
pp. 3209-3227
Author(s):  
Miriam Prume ◽  
Astrid Rollenhagen ◽  
Rachida Yakoubi ◽  
Kurt Sätzler ◽  
Joachim Hr Lübke
Keyword(s):  
Structural Parameters ◽  
Three Dimensional ◽  
Wild Type Mouse ◽  
Fine Scale ◽  
Synaptic Organization ◽  
Wild Type ◽  
Structural Determinants ◽  
Electron Microscopic ◽  
Synaptic Boutons ◽  
Active Zones

Abstract Synapses are key structural determinants for information processing and computations in the normal and pathologically altered brain. Here, the quantitative morphology of excitatory synaptic boutons in the “reeler” mutant, a model system for various neurological disorders, was investigated and compared with wild-type (WT) mice using high-resolution, fine-scale electron microscopy (EM) and quantitative three-dimensional (3D) models of synaptic boutons. Beside their overall geometry, the shape and size of presynaptic active zones (PreAZs) and postsynaptic densities (PSDs) forming the active zones and the three pools of synaptic vesicles (SVs), namely the readily releasable pool (RRP), the recycling pool (RP), and the resting pool, were quantified. Although the reeler mouse neocortex is severely disturbed, no significant differences were found in most of the structural parameters investigated: the size of boutons (~3 μm2), size of the PreAZs and PSDs (~0.17 μm2), total number of SVs, and SVs within a perimeter (p) of 10 nm and p20 nm RRP; the p60 nm, p100 nm, and p60–p200 nm RP; and the resting pool, except the synaptic cleft width. Taken together, the synaptic organization and structural composition of synaptic boutons in the reeler neocortex remain comparably “normal” and may thus contribute to a “correct” wiring of neurons within the reeler cortical network.

Quantitative Three-Dimensional Reconstructions of Excitatory Synaptic Boutons in Layer 5 of the Adult Human Temporal Lobe Neocortex: A Fine-Scale Electron Microscopic Analysis

2018 ◽  
Vol 29 (7) ◽  
pp. 2797-2814 ◽  
Author(s):  
Rachida Yakoubi ◽  
Astrid Rollenhagen ◽  
Marec von Lehe ◽  
Yachao Shao ◽  
Kurt Sätzler ◽  
...  
Keyword(s):  
Temporal Lobe ◽  
Three Dimensional ◽  
Microscopic Analysis ◽  
Synaptic Cleft ◽  
Brain Regions ◽  
Fine Scale ◽  
Electron Microscopic ◽  
Readily Releasable Pool ◽  
Synaptic Boutons ◽  
Human Temporal Lobe

Abstract Studies of synapses are available for different brain regions of several animal species including non-human primates, but comparatively little is known about their quantitative morphology in humans. Here, synaptic boutons in Layer 5 (L5) of the human temporal lobe (TL) neocortex were investigated in biopsy tissue, using fine-scale electron microscopy, and quantitative three-dimensional reconstructions. The size and organization of the presynaptic active zones (PreAZs), postsynaptic densities (PSDs), and that of the 3 distinct pools of synaptic vesicles (SVs) were particularly analyzed. L5 synaptic boutons were medium-sized (~6 μm2) with a single but relatively large PreAZ (~0.3 μm2). They contained a total of ~1500 SVs/bouton, ~20 constituting the putative readily releasable pool (RRP), ~180 the recycling pool (RP), and the remainder, the resting pool. The PreAZs, PSDs, and vesicle pools are ~3-fold larger than those of CNS synapses in other species. Astrocytic processes reached the synaptic cleft and may regulate the glutamate concentration. Profound differences exist between synapses in human TL neocortex and those described in various species, particularly in the size and geometry of PreAZs and PSDs, the large RRP/RP, and the astrocytic ensheathment suggesting high synaptic efficacy, strength, and modulation of synaptic transmission at human synapses.

A Relatively Inexpensive Microprocessor-Linked Digital Plamimeter for Electron Microscopic Morphometry

1981 ◽  
Vol 39 ◽  
pp. 266-269
Author(s):  
Lee D. Peachey
Keyword(s):  
Theoretical Basis ◽  
Structural Parameters ◽  
Three Dimensional ◽  
Structural Data ◽  
Large Set ◽  
Dimensional Electron ◽  
Electron Microscopic ◽  
Morphometric Methods ◽  
Electron Microscopic Morphometry ◽  
Separate Step

Stereology provides a theoretical basis for powerful morphometric methods for the estimation of three-dimensional structural parameters from two-dimensional electron micrographs of cells and tissues. These methods assume at the start that one has a sufficiently large set of micrographs containing valid structural data. The task of obtaining from these micrographs the large quantity of data needed to get statistically valid results has been eased in two general ways. Sampling of data in the micrograph can be done rapidly by point and intersection counting methods. An alternate method, planimetry, obtains all the data in the micrograph, but in general is more time-consuming than point and intersection counting. Some of the relative inefficiency of planimetry is compensated when a digital planimeter is coupled with a computer. Areas and lengths can be computed simultaneously as fast as profiles are traced. Furthermore, rapid and numerically accurate compilation and statistical analysis of the data can be done automatically as the planimetry is done, not as a separate step after the data have been obtained.

scholarly journals 3D Ultrastructural Study of Synapses in the Human Entorhinal Cortex

2020 ◽  
Vol 31 (1) ◽  
pp. 410-425 ◽  
Author(s):  
M Domínguez-Álvaro ◽  
M Montero-Crespo ◽  
L Blazquez-Llorca ◽  
J DeFelipe ◽  
L Alonso-Nanclares
Keyword(s):  
Entorhinal Cortex ◽  
Focused Ion Beam ◽  
Functional Organization ◽  
Structural Parameters ◽  
Ion Beam ◽  
Three Dimensional ◽  
Ultrastructural Level ◽  
3D Analysis ◽  
Synaptic Organization ◽  
Health And Disease

Abstract The entorhinal cortex (EC) is a brain region that has been shown to be essential for memory functions and spatial navigation. However, detailed three-dimensional (3D) synaptic morphology analysis and identification of postsynaptic targets at the ultrastructural level have not been performed before in the human EC. In the present study, we used Focused Ion Beam/Scanning Electron Microscopy to perform a 3D analysis of the synapses in the neuropil of medial EC in layers II and III from human brain autopsies. Specifically, we studied synaptic structural parameters of 3561 synapses, which were fully reconstructed in 3D. We analyzed the synaptic density, 3D spatial distribution, and type (excitatory and inhibitory), as well as the shape and size of each synaptic junction. Moreover, the postsynaptic targets of synapses could be clearly determined. The present work constitutes a detailed description of the synaptic organization of the human EC, which is a necessary step to better understand the functional organization of this region in both health and disease.

scholarly journals Synaptic Organization of the Human Temporal Lobe Neocortex as Revealed by High-Resolution Transmission, Focused Ion Beam Scanning, and Electron Microscopic Tomography

2020 ◽  
Vol 21 (15) ◽  
pp. 5558
Author(s):  
Astrid Rollenhagen ◽  
Bernd Walkenfort ◽  
Rachida Yakoubi ◽  
Sarah A. Klauke ◽  
Sandra F. Schmuhl-Giesen ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
3D Models ◽  
Synaptic Organization ◽  
Experimental Animals ◽  
Synaptic Boutons ◽  
Active Zones ◽  
Em Tomography ◽  
Human Temporal Lobe

Modern electron microscopy (EM) such as fine-scale transmission EM, focused ion beam scanning EM, and EM tomography have enormously improved our knowledge about the synaptic organization of the normal, developmental, and pathologically altered brain. In contrast to various animal species, comparably little is known about these structures in the human brain. Non-epileptic neocortical access tissue from epilepsy surgery was used to generate quantitative 3D models of synapses. Beside the overall geometry, the number, size, and shape of active zones and of the three functionally defined pools of synaptic vesicles representing morphological correlates for synaptic transmission and plasticity were quantified. EM tomography further allowed new insights in the morphological organization and size of the functionally defined readily releasable pool. Beside similarities, human synaptic boutons, although comparably small (approximately 5 µm), differed substantially in several structural parameters, such as the shape and size of active zones, which were on average 2 to 3-fold larger than in experimental animals. The total pool of synaptic vesicles exceeded that in experimental animals by approximately 2 to 3-fold, in particular the readily releasable and recycling pool by approximately 2 to 5-fold, although these pools seemed to be layer-specifically organized. Taken together, synaptic boutons in the human temporal lobe neocortex represent unique entities perfectly adapted to the “job” they have to fulfill in the circuitry in which they are embedded. Furthermore, the quantitative 3D models of synaptic boutons are useful to explain and even predict the functional properties of synaptic connections in the human neocortex.

Freeze-Fracture Replication in the Ultrastructure of Blue-Green Algae

1967 ◽  
Vol 25 ◽  
pp. 74-75
Author(s):  
L. V. Leak
Keyword(s):  
Cell Wall ◽  
Electron Microscope ◽  
Green Algae ◽  
Three Dimensional ◽  
Freeze Fracture ◽  
Electron Microscopic ◽  
Photosynthetic Membranes ◽  
Blue Green Algae ◽  
Cell Biol ◽  
Cellular Components

Electron microscopic observations of freeze-fracture replicas of Anabaena cells obtained by the procedures described by Bullivant and Ames (J. Cell Biol., 1966) indicate that the frozen cells are fractured in many different planes. This fracturing or cleaving along various planes allows one to gain a three dimensional relation of the cellular components as a result of such a manipulation. When replicas that are obtained by the freeze-fracture method are observed in the electron microscope, cross fractures of the cell wall and membranes that comprise the photosynthetic lamellae are apparent as demonstrated in Figures 1 & 2.A large portion of the Anabaena cell is composed of undulating layers of cytoplasm that are bounded by unit membranes that comprise the photosynthetic membranes. The adjoining layers of cytoplasm are closely apposed to each other to form the photosynthetic lamellae. Occassionally the adjacent layers of cytoplasm are separated by an interspace that may vary in widths of up to several 100 mu to form intralamellar vesicles.

Structural Role of rRNAs on the Ribosomal Subunits from E. Coli by Scanning Transmission Electron Microscopy

1981 ◽  
Vol 39 ◽  
pp. 424-425
Author(s):  
M. Boublik ◽  
N. Robakis ◽  
J.S. Wall
Keyword(s):  
Three Dimensional ◽  
Uranyl Acetate ◽  
Dimensional Structure ◽  
Electron Microscopic ◽  
Ribosomal Subunits ◽  
E Coli ◽  
Freeze Dried ◽  
16S Rna ◽  
Scanning Transmission ◽  
And Function

The three-dimensional structure and function of biological supramolecular complexes are, in general, determined and stabilized by conformation and interactions of their macromolecular components. In the case of ribosomes, it has been suggested that one of the functions of ribosomal RNAs is to act as a scaffold maintaining the shape of the ribosomal subunits. In order to investigate this question, we have conducted a comparative TEM and STEM study of the structure of the small 30S subunit of E. coli and its 16S RNA.The conventional electron microscopic imaging of nucleic acids is performed by spreading them in the presence of protein or detergent; the particles are contrasted by electron dense solution (uranyl acetate) or by shadowing with metal (tungsten). By using the STEM on freeze-dried specimens we have avoided the shearing forces of the spreading, and minimized both the collapse of rRNA due to air drying and the loss of resolution due to staining or shadowing. Figure 1, is a conventional (TEM) electron micrograph of 30S E. coli subunits contrasted with uranyl acetate.

EMCAT: An automatic image-processing system for electron-microscopic tomography

1994 ◽  
Vol 52 ◽  
pp. 418-419
Author(s):  
Weiping Liu ◽  
John W. Sedat ◽  
David A. Agard
Keyword(s):  
Image Processing ◽  
Structural Information ◽  
Imaging Technique ◽  
Three Dimensional ◽  
Processing System ◽  
Electron Microscopic ◽  
Data Set ◽  
Ordered Structures ◽  
Automatic Image Processing ◽  
Em Tomography

Any real world object is three-dimensional. The principle of tomography, which reconstructs the 3-D structure of an object from its 2-D projections of different view angles has found application in many disciplines. Electron Microscopic (EM) tomography on non-ordered structures (e.g., subcellular structures in biology and non-crystalline structures in material science) has been exercised sporadically in the last twenty years or so. As vital as is the 3-D structural information and with no existing alternative 3-D imaging technique to compete in its high resolution range, the technique to date remains the kingdom of a brave few. Its tedious tasks have been preventing it from being a routine tool. One keyword in promoting its popularity is automation: The data collection has been automated in our lab, which can routinely yield a data set of over 100 projections in the matter of a few hours. Now the image processing part is also automated. Such automations finish the job easier, faster and better.

scholarly journals Brain capillary structures of schizophrenia cases and controls show a correlation with their neuron structures

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Rino Saiga ◽  
Masayuki Uesugi ◽  
Akihisa Takeuchi ◽  
Kentaro Uesugi ◽  
Yoshio Suzuki ◽  
...  
Keyword(s):  
Blood Vessels ◽  
Structural Parameters ◽  
Three Dimensional ◽  
Superior Temporal Gyrus ◽  
Anterior Cingulate ◽  
Dimensional Structure ◽  
Constant Diameter ◽  
The Mean ◽  
Cartesian Coordinate ◽  
Micrometer Scale

AbstractBrain blood vessels constitute a micrometer-scale vascular network responsible for supply of oxygen and nutrition. In this study, we analyzed cerebral tissues of the anterior cingulate cortex and superior temporal gyrus of schizophrenia cases and age/gender-matched controls by using synchrotron radiation microtomography or micro-CT in order to examine the three-dimensional structure of cerebral vessels. Over 1 m of cerebral blood vessels was traced to build Cartesian-coordinate models, which were then used for calculating structural parameters including the diameter and curvature of the vessels. The distribution of vessel outer diameters showed a peak at 7–9 μm, corresponding to the diameter of the capillaries. Mean curvatures of the capillary vessels showed a significant correlation to the mean curvatures of neurites, while the mean capillary diameter was almost constant, independent of the cases. Our previous studies indicated that the neurites of schizophrenia cases are thin and tortuous compared to controls. The curved capillaries with a constant diameter should occupy a nearly constant volume, while neurons suffering from neurite thinning should have reduced volumes, resulting in a volumetric imbalance between the neurons and the vessels. We suggest that the observed structural correlation between neurons and blood vessels is related to neurovascular abnormalities in schizophrenia.

scholarly journals Coupled Fluid–Solid Numerical Simulation for Flow Field Characteristics and Supporting Performance of Flexible Support Cylindrical Gas Film Seal

Aerospace ◽  
2021 ◽  
Vol 8 (4) ◽  
pp. 97
Author(s):  
Junfeng Sun ◽  
Meihong Liu ◽  
Zhen Xu ◽  
Taohong Liao ◽  
Xiangping Hu ◽  
...  
Keyword(s):  
Flow Field ◽  
Film Thickness ◽  
Fluid Dynamic ◽  
Structural Characteristics ◽  
Structural Parameters ◽  
Equivalent Stress ◽  
Three Dimensional ◽  
High Rotational Speed ◽  
Flexible Support ◽  
Flow Field Characteristics

A new type of cylindrical gas film seal (CGFS) with a flexible support is proposed according to the working characteristics of the fluid dynamic seal in high-rotational-speed fluid machinery, such as aero-engines and centrifuges. Compared with the CGFS without a flexible support, the CGFS with flexible support presents stronger radial floating characteristics since it absorbs vibration and reduces thermal deformation of the rotor system. Combined with the structural characteristics of a film seal, an analytical model of CGFS with a flexible wave foil is established. Based on the fluid-structure coupling analysis method, the three-dimensional flow field of a straight-groove CGFS model is simulated to study the effects of operating and structural parameters on the steady-state characteristics and the effects of gas film thickness, eccentricity, and the number of wave foils on the equivalent stress of the flexible support. Simulation results show that the film stiffness increases significantly when the depth of groove increases. When the gas film thickness increases, the average equivalent stress of the flexible support first decreases and then stabilizes. Furthermore, the number of wave foils affects the average foils thickness. Therefore, when selecting the number of wave foils, the support stiffness and buffer capacity should be considered simultaneously.

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