Electron Micrographs Of Serial Sections Research Articles

Gap junctions between the pedicles of macaque foveal cones

Cone terminals (“pedicles”) in the fovea of macaque retina were studied in electron micrographs of serial sections. Pedicles were sheathed in glia except for small (0.2 μm 2) fenestrations, 4.8 ± 1.7 per pedicle. At each fenestration the membranes of adjacent pedicles were contiguous and marked by an adherent junction, which in turn was invariably associated with gap junctions. There were 3.2 ± 1.4 gap junctions per adherent junction and thus, about fifteen gap junctions per pedicle. The gap junctions were small, 1.6 × 10 −3 ± 1.8 × 10 −3μm 2 ( mean ± SD) and were formed indiscriminately with all neighboring pedicles. An upper bound was estimated of 170 connexons per pedicle and thus a coupling conductance of 1.7 × 10 4pS . Available psychophysical data suggest that the junctions are uncoupled at high luminance. They may couple at lower luminance where spatial averaging would improve contrast sensitivity without cost to spatial acuity.

Computational model of the on-alpha ganglion cell receptive field based on bipolar cell circuitry.

The on-alpha ganglion cell in the area centralis of the cat retina receives approximately 450 synapses from type b1 cone bipolar cells. This bipolar type forms a closely spaced array (9 microns), which contributes from 1 to 7 synapses per b1 cell throughout the on-alpha dendritic field. Here we use a compartmental model of an on-alpha cell, based on a reconstruction from electron micrographs of serial sections, to compute the contribution of the b1 array to the on-alpha receptive field. The computation shows that, for a physiologic range of specific membrane resistance (9500-68,000 omega.cm2) and a linear synapse, inputs are equally effective at all points on the on-alpha dendritic tree. This implies that the electrotonic properties of the dendritic tree contribute very little to the domed shapes of the receptive field center and surround. Rather, these shapes arise from the domed distribution of synapses across the on-alpha dendritic field. Various sources of "jitter" in the anatomical circuit, such as variation in bipolar cell spacing and fluctuations in the number of synapses per bipolar cell, are smoothed by the overall circuit design. However, the computed center retains some minor asymmetries and lumps, due to anatomical jitter, as found in actual alpha-cell receptive fields.

Multifunctional features of a gastrodermal sensory cell inHydra: three-dimensional study

Computer-assisted, three-dimensional reconstructions of two gastrodermal sensory cells from transmission electron micrographs of serial sections of Hydra revealed a unipolar morphology with the nucleus near an apical cilium and a simple unbranched axon with a widened terminal. The sensory cells were similar in size and shape to a unipolar sensory cell isolated from macerated gastrodermis and examined with scanning electron microscopy. In thin sections, the cells were characterized by the presence of numerous dense-cored vesicles in the axon and its terminal. A few dense-cored vesicles were aligned at electron-dense synaptic foci in the axon terminal of the sensory cell, which formed an axo-axonal synapse with a nearby centrally located ganglion cell and a neuromuscular synapse with the basal myoneme of a digestive cell. The ganglion cell possessed a perikaryal cilium and a slender axon that extended adjacent to the sensory cell terminal, where it formed an en passant axo-axonal synapse in reciprocal arrangement with that of the sensory cell. In addition, the ganglion cell axon formed a neuromuscular synapse in sequence with the sensory cell axo-axonal synapse. The presence of a large number of neurosecretory-like granules, apical cilium and reciprocal interneuronal and neuromuscular synaptic loci suggests that this gastrodermal sensory cell, characterized ultrastructurally for the first time, represents a third type of multifunctional neuron in Hydra. Thus, Hydra may contain primitive stem-like neurons, which are sensory-motor and also function in both neurosecretion and neurotransmission.

Patterns of synaptic input on corticocortical and corticothalamic cells in the cat visual cortex. I. The cell body

Immunocytochemical and electron microscopic methods were used to examine the ultrastructure and synaptology of callosal and corticothalamic pyramidal cell somata in the cat visual cortex (area 17). Callosal and corticothalamic cells were labeled after injection of horseradish peroxidase (HRP) in the contralateral visual cortex or in the ipsilateral lateral geniculate nucleus. The synaptic relationship between each of the two populations of pyramidal cells and cells containing the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) was examined at the light and electron microscope level using the combined techniques of retrograde transport of HRP and GABA immunocytochemistry. We found that callosal and corticothalamic cell somata have an ultrastructure and synaptology that distinguishes them from each other. Reconstructions from electron micrographs of serial sections revealed that the vast majority of synapses (89-96%) on the cell body of pyramidal cells were formed by GABAergic axon terminals, and that within each population of pyramidal cells there was variability in the number and density of axosomatic synapses. Callosal pyramidal cells received a greater number and higher density of axosomatic synapses than corticothalamic cells. These data suggest that callosal cells receive more inhibition than corticothalamic cells at the level of their somata.

Dynamic structure of Golgi apparatus depending upon functional phases of secretion.

Ultrastructural changes in Golgi apparatus (GA) during various functional conditions were studied in the rat parotid glands, and their three dimensional configuration was analyzed with the use of computer graphics from serial section electron micrographs. Three dimensional reconstruction confirmed that a normal GA was a connected single structure located in the supranuclear region, although it exhibited markedly complicated and reticulated branches. In contrast when host cells entered into the mitotic phase, underwent postnatal development and were inhibited of transport of secretory materials from rough endoplasmic reticulum, GA became an aggrega-tion of small vesicles or tubules, in which cis and most likely medial, but not trans elements were included. From these observations, we propose a new hypothesis that a GA consists of two components, basic and functional ones. The basic component having small vesicles and tubules of cis and medial natures is the most primitive feature of GA that is recognized in such cellular phases as early development, mitosis, and interruption of inflow. Basic components evolve a functional one that consists of a lamellar structure with cis, medial and trans elements. They are recognized in the usual cells at the active states. The size and overall configuration of GA changes greatly due to the amount of inflow of secretory materials depending upon the functional conditions of the cells.

Convergence and divergence of cones onto bipolar cells in the central area of cat retina

In the central area of cat retina the cone bipolar cells that innervate sublamina b of the inner plexiform layer comprise five types, four with narrow dendritic fields and one with a wide dendritic field. This was shown in the preceding paper (Cohen & Sterling 1990 a) by reconstruction from electron micrographs of serial sections. Here we show by further analysis of the same material that the coverage factor (dendritic spread x cell density) is about one for each of the narrow-field types (b1, b2, and b4). The same is probably true for the other narrow-field type (b3), but this could not be proved because its dendrites were too fine to trace. The dendrites of types b1, b2, and b4 collect from all the cone pedicles within their reach and do not bypass local pedicles in favour of more distant ones. The dendrites of type b5, the wide-field cell, bypass many pedicles. On average 5.1 +/- 1.0 pedicles coverage on a b1 bipolar cell; 6.0 +/- 1.2 converge on a b2 cell and 5.7 +/- 1.5 converge on a b4 cell. Divergence within a type is minimal: one pedicle contacts only 1.2 b1 cells, 1.0 b2 cells, and 1.0 b4 cells. Divergence across types is broad: each pedicle apparently contacts all four types of the narrow-field bipolar cells that innervate sublamina b. Each pedicle probably also contacts an additional 4-5 types of narrow-field bipolar cell that innervate sublamina a. There are several possible advantages to encoding the cone signal into multiple, parallel, narrow-field pathways.(ABSTRACT TRUNCATED AT 250 WORDS)

Dynamics of cytoplasmic organelles in the cell cycle of the fission yeast Schizosaccharomyces pombe: three-dimensional reconstruction from serial sections

Changes in the ultrastructure of the fission yeast Schizosaccharomyces pombe during the cell division cycle were analyzed by three-dimensional reconstruction of serial section electron micrographs of freeze-substituted cells. Cytoplasmic vesicles were found at the cell ends during interphase and at the equatorial zone of cells undergoing cytokinesis. Filasomes behaved in a similar but temporally retarded way to vesicles. Microfilament(mf)-associated granules were found attached to the plasma membrane at the growing ends. Microfilaments were identified against the plasma membrane and adjacent to developing septa. From these observations it is suggested that mf-associated structures such as filasomes constitute dense knots of actin network that function in localized cell wall growth by controlling the deposition of cytoplasmic vesicles. Dictyosomes occur as tubular and fenestrated cisternae with associated cytoplasmic vesicles. They were distributed uniformly in the cytoplasm and did not change significantly during the cell cycle. Changes in the three-dimensional localization of cytoplasmic microtubules and mitochondria are also described.

The role of polyfusomes in generating branched chains of cystocytes during Drosophila oogenesis

Three-dimensional models were constructed utilizing the information gained from electron micrographs of serial sections of two clones of cystocytes undergoing their terminal divisions. In each clone a polyfusome connected all eight cystocytes together. Each of the spindles was oriented so that one pole touched the polyfusomes, while the other pointed away from it. This positioning of spindles ensures that one cell of each dividing pair retains all previously formed canals, while the other receives none. The two cells that eventually come to contain the maximum number of canals and fusomal material are the ones that differentiate as pro-oocytes, while the others become nurse cells. The orientation of each spindle suggests that the polyfusome formed at one division determines the placement of the cytoskeletal fibers that anchor the spindles formed at the next division. There is a centripetal gathering together of new canals following each cycle of cystocyte division, which is thought to result from the subsequent contraction of the polyfusomal system. Females homozygous for the otu1 mutation are characterized by ovarian tumors, which result when germarial cystocytes undergo supernumerary divisions and fail to differentiate into either nurse cells or oocytes. An analysis of electron micrographs taken of serially sectioned, mutant germaria showed that most germ cells were single or belonged to clusters of two or three interconnected cells. Therefore otu1 cystocytes are unable to undergo a sustained series of arrested cleavages. These cystocytes contain fusomal material that shows ultrastructural differences from normal polyfusomes. We conclude: 1) that a normal polyfusomal system is a necessary prerequisite for the production of a branched chain of cystocytes and for their subsequent differentiation into pro-oocytes and nurse cells; and 2) that a product encoded by the otu+ gene is essential for the construction of a functional polyfusome.

The ON-alpha ganglion cell of the cat retina and its presynaptic cell types

Anatomical circuits converging onto the ON-alpha (Y) ganglion cell were studied by computer-assisted reconstruction of substantial portions of 2 alpha cells from electron micrographs of serial sections. The alpha cells in the area centralis were labeled by a Golgi-like retrograde filling with horseradish peroxidase, and certain presynaptic amacrine processes were labeled by uptake of 3H-glycine. About 4400 synapses contacted the alpha cell. Eighty-six percent were from amacrine cells; the rest were from bipolar cells. About one-quarter of the amacrine synapses were specifically labeled by 3H-glycine and probably belong to the A4 amacrine. The bipolar inputs were provided by several types: cone bipolar CBb1 (85%), cone bipolar CBb5 (2%), the rod bipolar (5%), and some unidentified cone bipolars (11%). Contacts from each type occurred in specific strata, with the consequence that they tended to form spots or annulli over the alpha dendritic field. The CBb1 bipolars formed a moderately dense array (8000/mm2), with a nearest-neighbor distance of 8.6 +/- 1.3 microns. Most members of the array (84%) contacted the alpha cell, providing 1-7 synapses (average, 2.7 +/- 1.6). The placement of contacts from an individual CBb1 followed certain rules: they were restricted to a parent branch of the alpha arbor or to 2 daughter branches, but almost never crossed a branch of the alpha arbor. The synaptic territory of an individual CBb1 was not shared with other b1s (or cone bipolars of any sort), although it was shared with amacrine contacts. Rod bipolar cells also formed a very dense array (54,500/mm2) in the alpha dendritic field, but only a few of these (3%) contacted the alpha cell. The concentric receptive field of the CBb1, combined with the spatial organization of its array, is used to predict the CBb1 contribution to the alpha cell receptive field; this contribution resembles the spatial and temporal organization of the alpha receptive field itself.

Synapses on dendritic shafts exhibit a perforated postsynaptic density

Synapses on dendritic shafts were examined in electron micrographs of serial sections obtained from the molecular layer of the rat dentage gyrus. Some of these synapses have been found to exhibit profiles of a discontinuous postsynaptic density (PSD). PSD reconstructions from serial sections were performed in a plane perpendicular to that of the sections. The results obtained indicate that profiles of discontinuous PSDs observed in random sections of dendritic shaft synapses are generated by sectioning of PSD plates that contain 1–3 holes or perforations. Earlier serial studies of osmicated material have demonstrated that a proportion of axospinous synapses also exhibit a perforated PSD. It appears, therefore, that the presence of PSD perforations is a general phenomenon shared by subpopulations of different types of synapses, both those involving dendritic shafts and those involving dendritic spines.

Tunneling cell processes in myocytes of stretched mouse atria.

Serial section electron micrographs of mouse atria stretched in vitro show that myocytes have cell processes which tunnel into adjacent myocytes for 8 microns or more. The tunneling cell processes (TCP) (diam 4-6.2 microns) lack myofibrils and organelles associated with atrial peptide secretion. The glycogen-rich TCP cytoplasmic matrix contains conspicuous tubules and vesicles originating from endoplasmic reticulum and resembling free sarcoplasmic reticulum (SR). TCP are surrounded by a plasmalemma derived from their myocyte of origin, the plasmalemma of the tunneled myocyte, and an intervening narrow compartment continuous with the interstitial space. Profiles having the characteristics cytoplasmic structure of TCP are also found both in the interstitial space between myocytes and near the longitudinal terminations where myocyte ends about on the interstitial space. We suggest that TCP tubules and vesicles may proliferate and/or transport in response to stretch, might be free SR, and may respond to stretch-activated changes in ionic composition or potential of the surrounding myocyte and narrow intercellular compartment.

The structure of the nervous system of the nematodeCaenorhabditis elegans

The structure and connectivity of the nervous system of the nematode Caenorhabditis elegans has been deduced from reconstructions of electron micrographs of serial sections. The hermaphrodite nervous system has a total complement of 302 neurons, which are arranged in an essentially invariant structure. Neurons with similar morphologies and connectivities have been grouped together into classes; there are 118 such classes. Neurons have simple morphologies with few, if any, branches. Processes from neurons run in defined positions within bundles of parallel processes, synaptic connections being made en passant. Process bundles are arranged longitudinally and circumferentially and are often adjacent to ridges of hypodermis. Neurons are generally highly locally connected, making synaptic connections with many of their neighbours. Muscle cells have arms that run out to process bundles containing motoneuron axons. Here they receive their synaptic input in defined regions along the surface of the bundles, where motoneuron axons reside. Most of the morphologically identifiable synaptic connections in a typical animal are described. These consist of about 5000 chemical synapses, 2000 neuromuscular junctions and 600 gap junctions.

Transfer of differentiated synaptic terminals from primary myotubes to new-formed muscle cells during embryonic development in the rat

Most fibres in adult muscles are derived from secondary myotubes which form around an earlier developing population of primary myotubes. We examined embryonic rat IVth lumbrical muscles at a stage when most secondary myotubes were less than 24 h old, in order to study their acquisition of synaptic terminals. Single nerve terminals were frequently seen to synapse simultaneously with a well-developed primary myotube and a new-formed secondary myotube. Reconstructions of serial section electron micrographs revealed that most of these shared terminals had synaptic specializations opposed to the primary myotube, but others appeared to transmit mainly to the secondary myotube. Primary myotubes are contacted by multiple nerve terminals before secondary myotube formation has begun, and we suggest that synaptic terminals are progressively transferred from primaries to secondaries, the observed shared terminals representing intermediate stages in this transfer.

Computer Reconstructions of Normal Human Alveoli From Serial Sections

Portions of two adjacent normal human alveoli were reconstructed from serial sections in order to examine normal alveolar organization, including anatomical relationships among the different cell types, the connective tissue matrix and gaps in the alveolar septum. Computer reconstructions were prepared from montaged electron micrographs of serial sections. Rotation of these reconstructions in the x, y or z axes allowed examination of the alveoli from many different aspects other than the actual plane of sectioning. Anatomical relationships “between Type I and Type II epithelial cells, alveolar macrophages, and pores of Kohn that could not he deduced from a single plane of the section (random sections) were revealed.

Interlocked bivalents in reconstructed metaphase I cells of bread wheat.

Complete reconstructions of all the bivalents were made from electron micrographs of serial sections through six pollen mother cells at metaphase I of meiosis in Triticum aestivum (hexaploid bread wheat). At least two of these metaphases contained interlocked pairs of bivalents. In one, two ring bivalents were interlocked, while in another a rod bivalent ran through the centre of a ring bivalent. Two other groups of bivalents were too closely appressed to allow separation into individual bivalents and may have contained interlocks. Meiosis in other anthers of the same plants examined by light microscopy was considered normal. The frequency of interlocking found was much higher than reported from light-microscope spreads. Not all interlocks in metaphase I cells need adversely affect meiosis, but knowledge of their regularity and form may facilitate understanding the processes of chromosome pairing.

The neural circuit for touch sensitivity in Caenorhabditis elegans

The neural pathways for touch-induced movement in Caenorhabditis elegans contain six touch receptors, five pairs of interneurons, and 69 motor neurons. The synaptic relationships among these cells have been deduced from reconstructions from serial section electron micrographs, and the roles of the cells were assessed by examining the behavior of animals after selective killing of precursors of the cells by laser microsurgery. This analysis revealed that there are two pathways for touch-mediated movement for anterior touch (through the AVD and AVB interneurons) and a single pathway for posterior touch (via the PVC interneurons). The anterior touch circuitry changes in two ways as the animal matures. First, there is the formation of a neural network of touch cells as the three anterior touch cells become coupled by gap junctions. Second, there is the addition of the AVB pathway to the pre-existing AVD pathway. The touch cells also synapse onto many cells that are probably not involved in the generation of movement. Such synapses suggest that stimulation of these receptors may modify a number of behaviors.

Light microscope based analysis of three-dimensional structure: applications to the study of Drosophila salivary gland nuclei. I. Data collection and analysis.

Many biological structures of interest are large enough that they may be viewed by light microscope methods, yet they are sufficiently complicated that interpretation of what is seen is quite difficult. The salivary gland nuclei from Dipterans are an example of this. Previous attempts at determining the path of the giant chromosomes in these nuclei have depended on the laborious construction of models by hand. A unified Computer Aided Modelling and Analysis system (CAMA) has been implemented, allowing data collection and analysis of structures visible by light microscopy. This system is extendible to the analysis of electron micrographs of serial sections or of other data consisting of images present in a stack.

Chromosome order – possible implications for development

ABSTRACT Chromosomes are arranged in ordered haploid sets around the centre of the metaphase plate at mitosis in several grass species and hybrids. Each chromosome is in a fixed mean position relative to other, heterologous chromosomes, this order can be predicted using Bennett’s model, and is clearly demonstrated from reconstructions of electron micrographs of serial sections (see Heslop-Harrison & Bennett, 1983a,b,c). The nucleus contains spatial domains of genes with similar functions. Chromosomes with major effects on nuclear behaviour - division or meiotic pairing - may be at special positions in the order. Changing spatial relationships of chromosomes with respect both to each other and the nuclear envelope (during the cell cycle and during development) may affect cell differentiation and gene activity. Chromosome order may have implications for the control of development within the nucleus and the organism. Order may constrain karyotype and hence species evolution.

An extensive lysosomal system in the maternal epithelium of the porcine placenta

The material comprised 64 placentae from 19 Danish Landrace sows representing gestational stages from 16 to 112 days. The maternal epithelium was examined by means of light and electron microscopy to describe the morphology of the irregular electron-dense bodies and membrane whorls and their relation to gestational stages. Incubation for acid phosphatase demonstrated their lysosomal nature. Electron micrographs of serial sections followed by reconstructions and freeze-fracturing demonstrated their three-dimensional structure and relation to transfer tubules. In the early stages, during the development of fetal vascularization, the maternal epithelium was dominated by inclusions of lipids and glycogen, which can easily be transferred to and utilized by the embryo. The development of an extensive lysosomal system, consisting of irregular electron-dense bodies, membrane whorls and transfer tubules, indicates that cellular digestion is an important factor in the transfer and subsequent fetal utilization of maternal nutriments in the last two-thirds of gestation, and that this development is apparently related to anatomical and physiological changes during gestation.

W10BSmL, a mutant of Euglena gracilis var. bacillaris lacking plastids.

Organized proplastid structures are absent from dark-grown and light-grown cells of Euglena gracilis Klebs var. bacillaris Cori mutant W10BSmL, based on electron micrographs of serial sections of entire cells. Fluorescence due to normal plastid DNA is undetectable in these cells after treatment with the DNA fluorochrome 4'6-diamidino-2-phenylindole (DAPI). Serial sections through a newly described compartmentalized osmiophilic structure in Euglena cells are presented.