Analysis Of Dendritic Spines Research Articles

Automated Three-Dimensional Detection and Shape Classification of Dendritic Spines from Fluorescence Microscopy Images

A fundamental challenge in understanding how dendritic spine morphology controls learning and memory has been quantifying three-dimensional (3D) spine shapes with sufficient precision to distinguish morphologic types, and sufficient throughput for robust statistical analysis. The necessity to analyze large volumetric data sets accurately, efficiently, and in true 3D has been a major bottleneck in deriving reliable relationships between altered neuronal function and changes in spine morphology. We introduce a novel system for automated detection, shape analysis and classification of dendritic spines from laser scanning microscopy (LSM) images that directly addresses these limitations. The system is more accurate, and at least an order of magnitude faster, than existing technologies. By operating fully in 3D the algorithm resolves spines that are undetectable with standard two-dimensional (2D) tools. Adaptive local thresholding, voxel clustering and Rayburst Sampling generate a profile of diameter estimates used to classify spines into morphologic types, while minimizing optical smear and quantization artifacts. The technique opens new horizons on the objective evaluation of spine changes with synaptic plasticity, normal development and aging, and with neurodegenerative disorders that impair cognitive function.

3-D confocal laser scanning microscopy used in morphometric analysis of rat Purkinje cell dendritic spines after chronic ethanol consumption.

A confocal laser scanning microscope (with a 543 nm laser) was used for imaging rat Purkinje cell dendritic spines at high 3-D resolution. In a nutritionally controlled study of the rat, 5 months of ethanol consumption was demonstrated to alter the spines of Purkinje cell dendrites in rat cerebellum. Intact spines showed significant elongation after ethanol exposure, whereas this neuromorphological alteration could not be detected in controls. Spine elongation could be regarded as compensative growth of spines in search of new synaptic contacts due to alcohol induced cell loss.

Analysis of dendritic spines in rat CA1 pyramidal cells intracellularly filled with a fluorescent dye

The dendritic branching pattern and the distribution of dendritic spines were studied in hippocampal neurones with an improved technique. In slices taken from adult Wistar rats, CA1 pyramidal cells were filled with Lucifer yellow and examined under a laser-scanning confocal microscope. The basal dendrites were found evenly distributed inside a regular cupola-shaped volume. Their total length was about 4,500 microns. The branches divided between one and three times, with the initial segments comprising less than 2%, and the long terminal segments (mean length, 119 microns) including more than 80% of the total length of the basal dendrites. The apical dendritic branches emerged obliquely from the main shaft, ran for a distance of 50 to 250 microns, and made up a total length of about 5,100 microns in stratum radiatum and between 1,100 and 3,200 microns in stratum lacunosum-moleculare. The mean total length of the dendritic tree was 11,900 microns. All values were corrected for shrinkage. Shrinkage was measured in three dimensions and was 20.2% in the horizontal (x/y) plane and 40.9% in the vertical (z) plane. Both the basal and the apical dendritic branches were covered by regularly spaced spines. When corrected for dehydration-induced shrinkage and for hidden spines, the density was 1.80 and 2.00 spines/microns dendritic length for the basal and apical dendritic branches, respectively. Apart from the initial parts of the branches, which had few or no spines, the spines were remarkably evenly spaced. In particular, the distance between spine heads was significantly different from a random distribution, suggesting a regulatory process for the spacing of spines.

Three‐dimensional organization of neuronal and glial processes: High voltage electron microscopy

Neurons and glia cells in the mammalian central nervous system have many complicated processes. They are too fine for light microscopic study and too complicated and widely spread for thin section electron microscopy. High-voltage electron microscopic (HVEM) stereo observation of thick Golgi preparation provides detailed 3-D images of their processes. Three-dimensional fine structures of astocytic processes in the neuropile and on the surface of neuronal somata, and those of the ruffed cell axon initial segment and thorny excrescences of CA3 pyramidal cell dendrites, are elucidated with the aid of HVEM stereoscopy of thick Golgi preparations. In addition, some results obtained by 3-D morphometrical analysis of dendritic spines using HVEM stereo images are shown. The examples presented here clearly show the usefulness of high-voltage electron microscope stereo observation of thick specimens for detailed morphological and morphometric study of the central nervous system.

Quantitative analysis of dendritic spines of pyramidal neurons in layer V of the sensomotor cortex of rats exposed on the ?Kosmos-1667? biosatellite

Quantitative analysis of dendritic spines of pyramidal neurons in layer V of the sensomotor cortex of rats exposed on the ?Kosmos-1667? biosatellite

Effect of chronic ethanol consumption on the fine structure of the dentate gyrus in long-sleep and short-sleep mice

The effect of short-and long-term chronic ethanol consumption on the fine structure of the dentate gyrus was examined in two lines of mice selected for their differential sensitivity to acute ethanol administration. Quantitative electron microscopic analysis of dendritic spines, axon terminals, and synaptic appositions revealed significant differences between the long-sleep and short-sleep mice. In control preparations, long-sleep mice were found to have larger spine areas and perimeters, larger axon terminals, and longer synaptic appositions than short-sleep mice. In addition, the shape of dendritic spines in the long-sleep mice was significantly more complex than those of short-sleep mice. Ethanol tended to increase this complexity in long-sleep mice only. Ethanol had only a limited effect on the other anatomical measures. The results provide evidence for ultrastructural differences between the nervous systems of these lines of mice which may have a role in their differential sensitivity to acute ethanol administration.

Three-dimensional analysis of dendritic spines. II. Spine apparatus and other cytoplasmic components.

A total of 342 dendritic spines (193 from the visual and 149 from the cerebellar cortex of the mouse) were analyzed in serial and several hundred of thousands of them in single sections, with respect to the presence and organization of the spine apparatus and other cytoplasmic components. The continuity of the spine apparatus with the smooth endoplasmic reticulum of the dendritic trunk was shown in three-dimensional reconstructions. The dense material of the spine apparatus was divided into "inner dense plate" and "outer dense plate". The close relationship between the outer dense plate and the postsynaptic density suggests that the spine apparatus functions as a postsynaptic protein synthesizing centre. The material from the outer dense plate could be used for a dynamic extension of the synaptic active zone. An extraspinous spine apparatus of the axon initial segment was partially reconstructed. Polyribosomes were found in all large spines of the visual cortex but were not so frequent in small spines and in Purkinje cell dendritic spines. Microfilamentous network and intermediate filaments occurred in the spines. The smooth endoplasmic reticulum of Purkinje cell dendritic spines was reconstructed. No spine apparatus and dense material were present in these spines.

Three-dimensional analysis of dendritic spines. III. Glial sheath.

The glial envelope of dendritic spines in the visual and cerebellar cortices was evaluated by analysis of serial sections. Three-dimensional reconstructions of the protoplasmic astrocyte processes were made and the quantitative proportions of the glial cover on dendritic spines were measured and compared. Whereas dendritic spines on spiny branchlets of Purkinje cells are, with the exception of afferent axon terminals, completely covered by the glial sheath (74.44%), dendritic spines of pyramidal cells are only partially covered (28.89%), so that spine stalks and even synaptic clefts frequently lack glial isolation. A new, relatively frequent configuration of subsurface cistern--astrocyte process--dendritic spine is described. A possible functional significance of the differences in the glial ensheathment of dendritic spines in visual and cerebellar cortices is discussed.

Brainstem immaturity in sudden infant death syndrome: A quantitative rapid Golgi study of dendritic spines in 95 infants

Quantitative analysis of reticular dendritic spines was performed on rapid Golgi impregnated neurons in 7 brainstem areas from 61 sudden infant death syndrome (SIDS) and 34 control infants. Throughout the first postnatal year, mean spine density in SIDS was significantly greater than the mean density in controls ( P < 0.0001). There were significantly higher values of spine density in SIDS compared to controls ( P < 0.0001) in both term and preterm infants. Within the SIDS brainstem itself, the density of dendritic spines was significantly different ( P < 0.05) between two medullary regions and between reticular and non-reticular formation areas. Among these brainstem areas in controls, there was no significant difference. Our findings indicate an immature developmental pattern of increased dendritic spine density in the SIDS brainstem which may be responsible for abnormal central respiratory and arousal control. These significant quantitative differences in spine density are considered in the present study to represent an anatomical substrate of brainstem immaturity in the multifactorial pathogenesis of SIDS.

Three-dimensional analysis of dendritic spines. I. Quantitative observations related to dendritic spine and synaptic morphology in cerebral and cerebellar cortices.

A total of 212 dendritic spines (108 from the visual and 104 from cerebellar cortices of the mouse) were analyzed in serial sections. Dendritic spines (DS) and synaptic active zones (SAZ) were classified according to their shape, and the following quantitative data were measured: DS stalk and bulb diameters, DS length and volume, number of cisterns of the spine apparatus, DS and SAZ surface areas and their mutual proportions. Quantitative relationships between the spine apparatus and the size of DS and SAZ, between the volume and surface area of DS and between the size of DS and the size of SAZ were studied. Thin, mushroom-shaped and stubby DS with simple (circular or oval), complex (perforated, annulate or horseshoe-shaped) and multifocal SAZ were found on terminal branches of pyramidal cell apical dendrites and club-shaped DS with simple (circular or oval) SAZ on spiny branchlets of Purkinje cells. Statistically significant differences were found between all values measured on various DS types in the visual cortex. Linear dependencies of the DS surface area on DS volume and of the SAZ surface area on the DS surface area were established. Only a limited area of DS plasma membrane (7-10%) was occupied by SAZ. This finding indicates a possible functional importance of the SAZ/DS (and possibly also of the total SAZ/total postsynaptic membrane) surface ratio.

Three-dimensional structure of dendritic spines in the rat neostriatum

Dendritic spines of rat neostriatal neurons were examined by light microscopy and high voltage stereo electron microscopy (HVEM) following selective staining by intracellular microinjection of horseradish peroxidase. Conventionally prepared material also was used for quantitative analysis of dendritic spines from serial thin sections of neostriatum. Stereo electron microscopy of semithin sections from rat neostriatum fixed using a protocol designed to preserve cytoskeletal integrity was employed to examine the organization of the dendritic spine cytoplasm. Light microscopic and HVEM examination of spiny dendrites and quantitative analysis of serial thin sections from normal material revealed no distinct spine types but rather continuous and independent variation of spine head diameter, stalk diameter, and stalk length. Likewise, there was no systematic relationship between any of these spine dimensions and dendritic diameter. Spine head membrane surface area was directly related to the area of the synaptic junctional membrane of the spine head. In semithin sections, the cytoplasm of the spine contained membranous saccules of spine apparatus and a delicate cytoskeletal network composed of microfilaments and a set of finer and more variable cytoskeletal filaments. It is proposed that this cytoskeletal network together with the spine apparatus is responsible for the maintenance and alteration of spine shape and in this way controls the effectiveness of axospinous synapses.