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R R Mize

Publications and source records attributed to R R Mize.

At least 55 records · Page 3Linked to original sources
PubMed1989

Enkephalin-like immunoreactivity in the cat superior colliculus: distribution, ultrastructure, and colocalization with GABA.

The distribution of enkephalin (ENK) immunoreactivity has been examined in the cat superior colliculus (SC) by means of light and electron microscope immunocytochemistry. The antisera were directed against leucine enkephalin but also recognized methionine enkephalin. Colocalization of ENK with gamma aminobutyric acid (GABA) was studied with a two-chromagen double-labeling technique. Enkephalin antiserum labeling was highly specific. Dense neuropil labeling was found only in a thin band 75-100 microns wide within the upper superficial gray layer of SC. Negligible neuropil labeling was seen deeper, except for patches of label within the intermediate gray layer. Intensely labeled neurons also had a specific distribution. Forty-seven percent were located within the upper 200 microns of SC, 40% within the deep superficial gray layer, 11% in the optic layer, and only 2% below that layer. Almost all ENK-labeled cells were small (mean area of 117 microns2). Some of these had horizontal fusiform cell bodies and horizontally oriented dendrites. Others had small round somata and thin, obliquely oriented dendrites. In double-labeling experiments, 18% of anti-ENK-labeled cells were also immunoreactive for GABA. Four distinct types of ENK-labeled profile were identified with the electron microscope. Presynaptic dendrites (PSD) with loose accumulations of synaptic vesicles were densely labeled with the antiserum. Conventional dendrites were also labeled. Both types of labeled profile received input from unlabeled synaptic terminals, including those from the retina that contained pale mitochondria and round synaptic vesicles and formed asymmetric synaptic contacts. Retinal terminals were never labeled with the antisera. However, some axon terminals with round synaptic vesicles, dark mitochondria, and symmetric synaptic densities were labeled by the antisera, as were some thinly myelinated axons. These results show that there is a small population of enkephalinergic neurons in the cat SC, some of which also contain GABA. Because not all cells with identical morphologies were double labeled, it appears that neurons of like morphology are chemically heterogeneous.

Animals↗
PubMed1989

Origin, distribution, and morphology of serotonergic afferents to the cat superior colliculus: a light and electron microscope immunocytochemistry study.

We have studied the serotonergic (5-HT) projection to the cat superior colliculus (SC) using serotonin antibody immunocytochemistry and retrograde transport of peroxidase-conjugated wheatgerm agglutinin (WGA-HRP). In 3 experiments, the two labels were combined in order to double label cells with both anti-5-HT and WGA-HRP. In the remaining experiments, the two labels were examined separately. Serotonin-like immunoreactive fibers were found throughout all layers of SC, but were most densely distributed within the zonal and upper superficial gray layers. Most 5-HT fibers were thin and had characteristic varicosities and terminal swellings. At the EM level, immunoreactive terminals and varicosities were found to contain small agranular vesicles and occasionally large granular vesicles (LGVs). Conventional synaptic densities were only rarely observed. Injections of WGA-HRP into SC resulted in labeling of neurons throughout the dorsal raphe nucleus and surrounding ventrolateral periaqueductal gray. Only a few cells were found in the raphe medianus and raphe pontis and none within the raphe magnus or other medullary raphe nuclei. Cells in the dorsal raphe giving rise to the SC projection varied in shape, size, and morphology and must represent more than one cell type. The morphology of these cells was indistinguishable from that of cells in the dorsal raphe which were double labeled by anti-5-HT and WGA-HRP. We conclude that the 5-HT innervation of the superior colliculus varies in density in different laminae, arises from several different cell types, and originates primarily from the dorsal raphe nucleus with minor projections from raphe medianus and raphe pontis.

Animals↗
PubMed1989

The neurons of the substantia nigra and zona incerta which project to the cat superior colliculus are GABA immunoreactive: a double-label study using GABA immunocytochemistry and lectin retrograde transport.

The neurotransmitter cytochemistry of neurons in the substantia nigra and zona incerta which project to the cat superior colliculus was examined. Neurons in both structures were double-labeled with an antibody to the transmitter GABA and a retrograde tracer, [3H]n-acetylated wheat germ agglutinin, injected into the superior colliculus. All cells in the zona incerta and substantia nigra which projected to the superior colliculus were labeled by the GABA antiserum. Most other neurons within the zona incerta and virtually all within the substantia nigra pars reticulata and pars lateralis were also labeled by the GABA antibody. By contrast, neurons in the substantia nigra pars compacta were not labeled by either the GABA antibody or wheat germ agglutinin. Nigrotectal cells in the substantia nigra were of medium to large size and most had stellate-shaped cell bodies. Zona incerta cells projecting to the superior colliculus were also of medium to large size, but most had horizontal fusiform cell bodies. This study demonstrates two new findings: (1) that all nigrotectal neurons in cat are immunoreactive to a GABA antibody and probably contain the neurotransmitter GABA; and (2) that these GABA immunoreactive neurons in cat are found not only in the substantia nigra pars reticulata but also within the pars lateralis. Zona incerta cells projecting to the superior colliculus have a different morphology but are also apparently GABAergic. These data provide an anatomical substrate for the known inhibitory action of the nigrotectal pathway on superior colliculus neurons.

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PubMed1989

Cat-301 antibody selectively labels neurons in the Y-innervated laminae of the cat superior colliculus.

Cat-301 is a monoclonal antibody which recognizes a cell surface associated antigen of selected neurons in the central nervous system (CNS). In the visual system, cat-301 selectively labels Y-like cells in several visual structures, including portions of the lateral geniculate nucleus complex and visual cortex. The cat superior colliculus (SC) also receives Y input and contains cells driven by Y input which are selectively distributed in the deep superficial gray and deeper laminae. If cat-301 is selective to the Y-cell system in SC, labeled cells should be restricted to those laminae. To test this hypothesis, we have examined quantitatively the laminar distribution, percentage, size, and morphology of cells in SC labeled by the cat-301 antibody. Cat-301 labeled a variety of cells in the cat SC. Labeled cells were found within the deep portion of the superficial gray layer (6.6%), optic layer (27.6%), intermediate gray layer (26.9%), and the deep gray and white layers (38.5%). By contrast, only 2 of 667 labeled cells (0.3%) were found within that part of the upper superficial gray layer innervated exclusively by W input and thought to contain only W-driven cells. When considered as a percentage of the total cell population, cat-301 labeled cells represented less than 3% of cells in the superficial gray layer and approximately 15% in the deeper layers. Neurons labeled by cat-301 were all of medium to large size (mean average diameter = 33.3 microns; range = 15-84 microns) and included vertical fusiform and stellate cells in the upper layers and the very large neurons found in the intermediate gray and deeper layers. These results provide further evidence that the cat-301 antibody selectively recognizes the Y channel of the cat visual system.

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PubMed1988

Immunocytochemical localization of gamma-aminobutyric acid (GABA) in the cat superior colliculus.

This paper reports the pattern of labeling in the cat superior colliculus produced by an antiserum raised against BSA-conjugated gamma aminobutyric acid (GABA) and visualized by light and electron microscope immunocytochemistry. Neuropil labeling was densest within the zonal and superficial gray layers but was also found in the deep layers. Neurons labeled by the GABA antibody were also most dense within the zonal and superficial gray layers, although many labeled neurons were also found in the deeper layers. The ratio of labeled to unlabeled cells varied from an average of 45% in the superficial subdivision and the intermediate gray layer to less than 30% in the deeper laminae. Almost all intensely labeled cells were small (mean area = 127 micron 2) and had varied morphologies. Several types of labeled cell were observed with the electron microscope. One type had a horizontal, fusiform cell body and a deeply invaginated nucleus. Another type had a small round or ovoid cell body with cytoplasm clumped at one end. Labeled cells with other morphologies were also occasionally seen. No labeled glial cells were found. Two types of vesicle-containing dendrite were stained by the GABA antibody. One type had loose accumulations of small synaptic vesicles and often received input from retinal terminals. Another type had spines also containing small synaptic vesicles. Labeled dendrites without synaptic vesicles were also seen frequently. Putative axon terminals labeled by the GABA antibody had densely packed synaptic vesicles and formed symmetric synaptic contacts. Labeled myelinated axons were also commonly found. These results confirm those using uptake of tritiated GABA (Mize et al.: J. Comp. Neurol. 202:385-396, '81, J. Comp. Neurol, 206:180-192, '82) in that two of the same classes of GABA neuron, horizontal I and granule I cells, were identified in the superficial laminae. However, the GABA antiserum used in this study also revealed a third class of GABA neuron with vesicle-containing spines. The antiserum also labeled a significant number of putative GABAergic neurons located in the deep subdivision of the cat superior colliculus which were not previously recognized by using transmitter autoradiography.

Animals↗
PubMed1988

Quantitative immunocytochemistry using an image analyzer. I. Hardware evaluation, image processing, and data analysis.

In this review we describe how video-based image analysis systems are used to measure immunocytochemically labeled tissue. The general principles underlying hardware and software procedures are emphasized. First, the characteristics of image analyzers are described, including the densitometric measure, spatial resolution, gray scale resolution, dynamic range, and acquisition and processing speed. The errors produced by these instruments are described and methods for correcting or reducing the errors are discussed. Methods for evaluating image analyzers are also presented, including spatial resolution, photometric transfer function, short- and long-term temporal variability, and measurement error. The procedures used to measure immunocytochemically labeled cells and fibers are then described. Immunoreactive profiles are imaged and enhanced using an edge sharpening operator and then extracted using segmentation, a procedure which captures all labeled profiles above a threshold gray level. Binary operators, including erosion and dilation, are applied to separate objects and to remove artifacts. The software then automatically measures the geometry and optical density of the extracted profiles. The procedures are rapid and efficient methods for measuring simultaneously the position, geometry, and labeling intensity of immunocytochemically labeled tissue, including cells, fibers, and whole fields. A companion paper describes non-biological standards we have developed to estimate antigen concentration from the optical density produced by antibody labeling (Nabors et al., 1988).

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PubMed1988

Quantitative immunocytochemistry using an image analyzer. II. Concentration standards for transmitter immunocytochemistry.

Image analyzers can measure both the optical density and geometry of immunocytochemically labeled cells and fibers, as reviewed in a companion paper (Mize et al., 1988). In this paper, we report a procedure which allows us to estimate the concentration of a neurotransmitter based upon the optical density of antibody labeling produced by immunocytochemistry. To accomplish this, we developed a standard which binds conjugated neurotransmitters. Several artificial media for the standard were compared, including agar, gelatin, and agar-gelatin. A 3% agar matrix was found to be most suitable because it cut well and was nearly transparent. The agar sections were activated with cyanogen bromide/acetonitrile to promote coupling to the antigen. To test the standard, we used gamma-aminobutyric acid (GABA) conjugated to bovine serum albumin (BSA) as the antigen. The antibody was directed against this conjugate. Activated agar sections were incubated in serial dilutions of the tritium-labeled GABA/BSA conjugate. The radioactivity of some of these sections was measured to estimate the amount of coupled antigen. The remaining sections were incubated in the GABA antibody and processed for immunocytochemistry. The optical density of these sections was measured with an image analyzer. A linear relationship was found between GABA concentration and optical density over a range of at least 0.01 to 1 nmol/mg of agar. These results show that the concentration of bound GABA can be estimated from the optical density of sections labeled by antibody immunocytochemistry. The applicability of this technique to fixed brain tissue is discussed.

Image Processing, Computer-Assisted↗
PubMed1988

Laminar organization and ultrastructure of GABA-immunoreactive neurons and processes in the dorsal lateral geniculate nucleus of the tree shrew (Tupaia belangeri).

The distribution and ultrastructure of neurons and neuropil labeled by an antiserum to gamma-aminobutyric acid (GABA) were examined in the lateral geniculate nucleus (LGN) of the tree shrew (Tupaia belangeri). The LGN of this species segregates center type and cell class into three distinct pairs of laminae: a medial pair (laminae 1 and 2) containing ON-center cells, a more lateral pair (4, 5) containing OFF-center cells, and 2 laminae (3, 6) containing W-like cells. The relationship between this laminar segregation and the distribution of GABA immunoreactivity was investigated in the present study. GABA-immunoreactive neurons and neuropil were present in all six of the laminae. However, both the density of labeled cells (adjusted for neuronal density across laminae) and the density of labeled neuropil showed a medial-to-lateral gradient. The adjusted density of labeled cells was higher laterally than medially, and the density of labeled neuropil was significantly greater in the more lateral OFF-center laminae and W-like laminae than in the medial two ON-center laminae. Thus, inhibitory, GABAergic influences may modulate to different degrees the visual signals in the ON, OFF, and W pathways. Labeled cells had a mean cross-sectional area (107 microns 2) approximately one-half that of unlabeled cells (216 microns 2). They constitute 16-34% of the neurons in the LGN. At the electron microscope level, three different kinds of labeled profile were observed. Vesicle containing profiles like the F2 profiles of cat were postsynaptic to retinal terminals and presynaptic to conventional dendrites. F1 axon terminals with dense clusters of vesicles were also labeled as were some myelinated axons. Another labeled profile, which we suggest should be called an F3 process, was a large dendrite of irregular caliber with punctate groups of vesicles near the synapse. Our results suggest that GABAergic circuitry is an important part of the functional organization in the LGN of the tree shrew.

Animals↗
PubMed1987

The innervation density of serotonergic (5-HT) fibers varies in different subdivisions of the cat lateral geniculate nucleus complex.

The innervation density of serotonin (5-HT)-immunoreactive fibers, identified using an antibody to 5-HT, was found to differ in the 4 subdivisions of the cat lateral geniculate nucleus complex (LGN). The mean density (fiber length per unit area) of anti-5-HT-stained fibers was highest in the ventral LGN (0.062 micron per micron 2), moderate in the medial interlaminar nucleus (MIN) and the parvicellular C laminae of the dorsal LGN (0.039-0.040 per micron 2), and lowest in the A and magnocellular C laminae of the dorsal LGN (0.020 per micron 2). The fiber density in MIN was particularly dense along the medial edge of the nucleus, a region called the geniculate wing. The heaviest serotonin innervation is thus found in geniculate structures receiving input from W-type retinal ganglion cells and lightest in structures receiving X and Y input.

Animals↗
PubMed1986

Quantitative comparison of retinal synapses in the dorsal and ventral (parvicellular) C laminae of the cat dorsal lateral geniculate nucleus.

Three physiological classes of retinal ganglion cell project to the cat dorsal lateral geniculate nucleus (DLGN). The dorsal laminae A, A1, and magnocellular C receive X and Y retinal input, whereas the ventral parvicellular laminae C1 and C2 receive predominantly W input. We have compared quantitatively the retinal synaptic terminals of the dorsal and ventral laminae to determine whether there are morphological differences in the terminals that correspond to their different response properties. Anterogradely labeled retinal synaptic terminals in all laminae contained pale mitochondria and large, round synaptic vesicles. However, retinal terminals with pale mitochondria varied in size and synaptic organization in different laminae. The terminals in the A laminae were, on average, quite large and made numerous contacts with conventional dendritic profiles and with profiles that themselves contained synaptic vesicles (F2 profiles). The terminals in lamina C that contained pale mitochondria had a smaller overall mean area. Terminals with pale mitochondria in C1 and C2 were almost all small and synapsed with F2 profiles less frequently than did terminals in the A laminae or in lamina C. These results provide quantitative evidence that visual areas receiving W-type retinal input contain smaller retinal terminals and have a different synaptic organization from that of laminae receiving X and Y input.

Animals↗
PubMed1984

Retinal synapses of the cat medial interlaminar nucleus and ventral lateral geniculate nucleus differ in size and synaptic organization.

The retinal terminals of the medial interlaminar nucleus (MIN) and ventral lateral geniculate nucleus ( VLG ) have been examined quantitatively to determine if there are morphological differences in their synaptic ultrastructure which reflect their distinctive physiologies . The cross-sectional area and density (number per unit area) of synaptic contact zones with conventional and presynaptic dendrites (F2 profiles) were measured for each retinal terminal. The densities of F2 presynaptic dendrites and F1 flattened vesicle axon terminals were also measured. Retinal terminals in MIN were often large (mean size = 2.7 micron2 area) and had a high density of synaptic contacts (0.14 per micron surface area) with conventional dendrites, presynaptic dendrites, and dendritic spines. A high density of F2 presynaptic dendrites (0.08 per micron2 area) was found in MIN. F1 axon terminals were also found frequently (0.04 per micron2). MIN retinal terminals were often organized in glomeruli like those of the dorsal lateral geniculate nucleus. The retinal terminals in VLG were almost always small (mean size = 0.94 micron2 area), although they also had a high density of synaptic contacts (0.17 per micron surface area). They frequently synapsed on small dendrites and dendritic spines and less frequently on large dendrites. Unlike MIN, retinal terminals in VLG rarely contacted F2 presynaptic dendrites which were much less frequent in VLG (0.01 per micron2 area). Like MIN, VLG contained numerous F1 axon terminals (0.06 per micron2 area). No typical retinal glomeruli were found in VLG . These results show that MIN, which contains many Y cells, has a population of large retinal terminals and many F2 presynaptic dendrites. VLG , which apparently has only W cells, contains only small retinal terminals and has fewer F2 presynaptic dendrites. Both have a high density of F1 flat vesicle axon terminals.

Animals↗
PubMed1983

Variations in the retinal synapses of the cat superior colliculus revealed using quantitative electron microscope autoradiography.

This study has examined the retinal synapses of the cat superior colliculus using electron microscope autoradiography and morphometric techniques. The depth of each retinal synapse was measured using a computer-based EM plotter. The area, perimeter, and synapse contact density of selected synapses were calculated using a computer-based digitizer. Pale mitochondria were found to be an accurate cytological marker of retinal input to the colliculus. Fifty-eight percent of pale mitochondria terminals were labeled in the colliculus contralateral to eye injections. Ten percent of pale mitochondria terminals were labeled in the ipsilateral colliculus. A few labeled terminals contained dark mitochondria. The labeled retinal terminals in the contralateral colliculus were concentrated in a 60 microns wide dense band at the top of the superficial gray layer. They were also found within the deep superficial gray and upper optic layers. This distribution corresponded exactly to a larger population of pale mitochondria terminals. The cross-sectional area and synaptic contact density of selected pale mitochondria terminals varied with depth. Within the upper superficial gray, the terminals were small (mean area = 1.26 microns squared) and had high contact densities (mean = 0.25 per microns). These small terminals were also found deeper within the colliculus. Below the upper subdivision of the superficial gray, some labeled terminals were much larger and had lower contact densities. These results suggest there may be two subpopulations of retinal terminal in the cat superior colliculus: (1) small terminals with scalloped contours and complex synaptic relationships which may correspond to W-type input; and (2) larger terminals with simpler synaptic relationships which are distributed deeper and may correspond to Y-type input.

Animals↗
PubMed1983

A simple microcomputer-based three-dimensional serial section reconstruction system (MICROS).

We have developed a computer system which enters and aligns serial sections and displays the completed reconstructions at different rotations in space. The system uses commercially available hardware, including a Hewlett-Packard 9845T microcomputer and an H-P 9874A digitizer. The software for the system is written in the BASIC language. The system consists of two programs, one for section digitization, the other for rotation and display of the reconstructions. Sections are digitized directly from micrographs or back-projected slides. The outlines of cells or other structures are traced from these media using a hand-held cursor on the digitizer. The positions of elements (inputs) which contact the structure and fiducials are also digitized. The sections are aligned by simultaneously displaying two consecutive sections on the graphics CRT screen. The sections are coarsely superimposed by centering around screen center using a centering algorithm. They are precisely aligned by rotating and translating the images with a reference cursor. Special functions for inserting and deleting sections and rapid section scanning are available for editing. The aligned sections are stored using a linked-list file structure on either floppy disks or tape cartridges. The rotation program replots the completed reconstructions on the graphics CRT or digital plotter. The program will reproduce the reconstructions at any scale and at any rotation in the x-, y- or z-planes. A hidden line algorithm removes hidden lines to give a 3-dimensional (3-D) perspective to the reconstructions. The positions of inputs and fiducials are represented by symbols. We use the system to reconstruct cells and neural processes. The 3-D reconstructions allow us to: (a) examine the spatial distribution and density of synaptic contacts on neurons; (b) study complex neuronal shapes; (c) examine the vectors of neural processes. The computer reconstruction system, which is moderately priced, should also prove useful for reconstructing many other types of biological profile.

Animals↗
PubMed1983

A computer electron microscope plotter for mapping spatial distributions in biological tissues.

We have designed a computer-based electron microscope plotting system which maps the locations of organelles in tissue specimens and analyzes their distribution. The system includes: (1) two optical incremental shaft encoders which translate stage drive rotation to electrical pulses; (2) a Display/Control unit used to convert encoder pulses to binary code for computer input; (3) two 16-bit parallel interfaces for transferring data to the computer; (4) a Hewlett-Packard 9845T microcomputer, used to control data input and to store, graph, and analyze the plots. The software for the plotter is written in enhanced BASIC. The plotter system is driven by 4 programs called Trace, Plot, Analyze, and Density. The Trace program "draws" an outline of the edges of the tissue. The Plot program maps the positions of profiles within the tissue. The Analyze program compares trace and plot data and calculates the depth and medial-lateral distance of each plotted profile from the surfaces of the tissue. The Density program sorts and counts profile types, measures surface areas, and calculates profile densities. Commercial statistical software is used to analyze the data. Our laboratory uses the system to map the spatial distribution of synapses and neurons in the central nervous system. The plotting system will also be of value in other areas of neurobiology research.

Animals↗
PubMed1983

A microcomputer system for measuring neuron properties from digitized images.

A microcomputer system has been developed for the quantitative analysis of neurons, synapses, and other biological profiles. The outlines of profiles are traced from micrographs using a digitizer. From these traces, the system calculates cross-sectional area, perimeter (length), average diameter, and form factor. Data values for the number of profile contacts, the number of elements within a profile, the number of elements associated with a profile, profile type, and profile depth are also entered. The program calculates contact and element densities from these values. The data are printed-out on a thermal printer and stored on magnetic tape or floppy disk. The software is written entirely in BASIC. The system uses commercially available hardware, including a Hewlett-Packard 9845T microcomputer and H-P 9874A digitizer. The programs can be adapted to a wide variety of other microcomputer systems.

Animals↗
PubMed1982

Two types of GABA-accumulating neurons in the superficial gray layer of the cat superior colliculus.

Two types of neuron in the upper superficial gray layer of the cat superior colliculus accumulated exogenous 3H-gamma-aminobutyric acid intensely. The first type was a horizontal cell with a fusiform cell body, horizontal dendrites, a low synaptic density, but a high percentage of cortical synaptic contacts. This cell had presynaptic dendrites. The second type was a granule cell (type A) with a small round cell body, thin and obliquely oriented dendrites, a moderate synaptic density, and few cortical synaptic contacts. These two types differed in size, shape, dendritic morphology, and patterns of synaptic input. They likely participate in different inhibitory mechanisms. Four types of unlabeled neurons were also identified. Type B granule cells were found only within the upper subdivision of the superficial gray layer. They had moderate-sized cell bodies, a high synaptic density, and numerous somatic spines. A third type of granule cell (type C) was found only in the deep subdivision of the superficial gray. This type had a low synaptic density and spines that contained synaptic vesicles. Vertical fusiform and stellate forms were also found. We conclude that at least six types of neurons populate the upper superficial gray layer of the cat superior colliculus.

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