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Biomedical subjects

J K Stevens

Publications and source records attributed to J K Stevens.

33 records · Page 2Linked to original sources

The effects of polycations on vascular permeability in the rat. A proposed role for charge sites.

This study investigated whether charge sites in the walls of the microvasculature may play a role in maintaining the impermeability of the nonrenal capillaries to albumin. All experiments were performed in nephrectomized rats, studied in the awake state. The intravenous injection of protamine sulfate (4 mg/100 g body wt dissolved in 0.9% saline) was followed by a mean increase of 29.1% in hematocrit and a decrease of 28.4% in plasma albumin concentration over a 10-min period, indicating a significant 50-60% loss of albumin from the vascular space; a finding confirmed by studies using exogenous 125I-labeled albumin. Changes persisted for the remaining 80 min of observation, and could be reproduced by the injection of two other polycations, hexadimethrine and poly-l-lysine. These effects were not prevented by the antihistamine diphenhydramine hydrochloride. In contrast to 125I-labeled albumin, 14C-labeled neutral dextran of comparable size was not confined to the vascular space; its apparent volume of distribution progressively increased during the 90 min of observation. Intravenous injection of protamine sulfate was followed by a significantly smaller loss of 14C-dextran (36.5%) than albumin (59.1%) from the vascular space (P less than 0.01). Protamine sulfate could not be demonstrated to result in any changes in the physicochemical characteristics of albumin. These observations suggest that the negative charge sites present in nonglomerular capillary walls have functions similar to equivalent sites present in the glomerular capillaries. Thus, charge sites could contribute to the low permeability of the microvasculature to negatively charged macromolecules such as albumin. This may be an important mechanism for retaining albumin in the vascular space and preventing edema formation in health.

Animals↗

Microcircuitry of bipolar cells in cat retina.

We have studied 15 bipolar neurons from a small patch (14 X 120 micron) of adult cat retina located within the area centralis. From electron micrographs of 189 serial ultrathin sections, the axon of each bipolar cell was substantially reconstructed with its synaptic inputs and outputs by means of a computer-controlled reconstruction system. Based on differences in stratification, cytology, and synaptic connections, we identified eight different cell types among the group of 15 neurons: one type of rod bipolar and seven types of cone bipolar neurons. These types correspond to those identified by the Golgi method and by intracellular recording. Those bipolar cell types for which we reconstructed three or four examples were extremely regular in form, size, and cytology, and also in the quantitative details of their synaptic connections. They appeared quite as specific in these respects as invertebrate "identified" neurons. The synaptic patterns observed for each type of bipolar neuron were complex but may be summarized as follows: the rod bipolar axon ended in sublamina b of the inner plexiform layer and provided major input to the AII amacrine cell. The axons of three types of cone bipolar cells also terminated in sublamina b and provided contacts to dendrites of on-beta and other ganglion cells. All three types, but especially the Cb1, received gap junction contacts from the AII amacrine cell. Axons of four types of cone bipolar cells terminated in sublamina a of the inner plexiform layer and contacted dendrites of off-beta and other ganglion cells. One of these cone bipolar cell types, CBa1, made reciprocal chemical contacts with the lobular appendage of the AII amacrine cell. These results show that the pattern of cone bipolar cell input to beta (X) and probably alpha (Y) ganglion cells is substantially more complex than had been suspected. At least two types of cone bipolar contribute to each type of ganglion cell where only a single type had been anticipated. In addition, many of the cone bipolar cell pathways in the inner plexiform layer are available to the rod system, since at least four types of cone bipolar receive electrical or chemical inputs from the AII amacrine cell. This may help to explain why, in a retina where rods far outnumber the cones, there should be so many types of cone bipolar cells.

Animals↗

Rings of cross-striated fibrils within the cat cone pedicle: a computer-assisted serial EM analysis.

Using serial electron micrographs and a computer reconstruction system the authors have examined the three-dimensional cytoarchitecture of the cat cone pedicle. These reconstructions reveal that within each pedicle is a closed 4-6 micron diameter ring of cross-striated fibrils similar to the nonring structures described in photoreceptor inner and outer segments by others in guinea pig, rat, chick, monkey, and humans. Our cone pedicle rings encompass the invaginating synaptic contacts of the cone bipolars and horizontal cells and have periodic 70 nm striations surrounding bundles of fibrils about 100 nm in diameter. The authors suggest that these striated rings may be active contractile elements and could be responsible for shape changes in cone pedicles during dark adaptation.

Animals↗

Serial reconstruction of microtubular arrays within dendrites of the cat retinal ganglion cell: the cytoskeleton of a vertebrate dendrite.

Serial reconstruction at the EM level of cat retinal ganglion cell dendrites reveals that: (1) the microtubular array is discontinuous, (2) microtubular endings are associated with smooth endoplasmic reticulum (SER), mitochondria, and plasma membrane, (3) individual microtubules always maintain a minimum distance from other microtubules (87 nm), SER (43 nm) and plasma membrane (69 nm), and (5) individual microtubules can 'wander' independent of adjacent microtubules throughout the dendritic volume. These observations, taken with some recent biochemical and immunohistochemical data by other workers, suggest that the microtubules are surrounded by a coat of high molecular weight, microtubular-associated proteins (HMW MAPs), which effectively creates a 90 nm tube around a central microtubular core. Our results suggest that bundles of these 'MAP-tubes' may serve as a major component of the dendritic cytoskeleton in the cat ganglion cells.

Animals↗

Oculoparalytic illusion: visual-field dependent spatial mislocalizations by humans partially paralyzed with curare.

In darkness, observers partially paralyzed with curare make large (greater thn 20 degrees) gaze- and dosage-dependent errors in visually localizing eye-level-horizontal and median planes, in matching the location of a sound to a light, and in pointing at a light. In illuminated, structured visual localization and pointing are accurate but errors in auditory-to-visual matches remain. Defects in extraretinal eye position information are responsible for all errors. The influence of extraretinal eye position information on visual localization is suppressed by a structured visual field but is crucial both in darkness and for intersensory localization if visual capture is prevented.

Auditory Perception↗

Microtubular disarray in cortical dendrites and neurobehavioral failure. II. Computer reconstruction of perturbed microtubular arrays.

A previous report details morphological alterations in dendritic structure of cortical neurons in severe neurobehavioral retardation of unknown etiology. Using computer graphic techniques, the present study describes perturbations in the 3-dimensional character of the microtubular array, which correspond to degenerative change in dendritic geometry. In large proximal processes, two types of array have been reconstructed. Segmented microtubules may form a continuous helical swirl which underlies a bulge in the dendritic cylinder. Alternatively, small groups of microtubules, while maintaining orderly internal organization, may be disoriented with respect to the long axis of the process. In varicose regions of the dendrite the microtubular array is discontinuous. Microtubules course side by side through constructed regions, only to splay out and terminate within expanded regions. These pathological alterations in the microtubular array contrast sharply with the cortical dendritic microtubular array reconstructed from the normal adult mouse. Perturbation in those parameters which determine packing of microtubules within the dendritic process is also documented. In the pathological condition, microtubules lose the ability to exclude one another from close approach. The role of cross-linking molecules in maintaining the integrity of the microtubular array, and the role of microtubules in maintaining the geometry of the dendrite, are considered.

Cerebral Cortex↗

The dendritic varicosity: a mechanism for electrically isolating the dendrites of cat retinal amacrine cells?

Amacrine dendritic varicosities from cat retina were reconstructed using serial electron micrographs. Each varicosity contained a synaptic input and a synaptic output, suggesting that they may function as isolated local circuits. A passive steady state electrical model demonstrated that for a given conductance change the varicose dendrite maximizes the local membrane potential and minimizes th distance membrane potential change as compared to other possible dendritic shapes. We, therefore, suggest that the function of the varicosities on amacrine cell dendrites might be to electrically isolate these local input-output circuits.

Animals↗

Toward a functional architecture of the retina: serial reconstruction of adjacent ganglion cells.

Twenty adjacent ganglion cells in cat retina were partially reconstructed from electron micrographs of serial thin sections. Cells were classified by size and by dendritic branching patterns as alpha, beta, or gamma cells. The alpha and beta cells were further subdivided by differences in the laminar distribution of their dendrites in the inner plexiform layer. The distribution of synaptic contacts on the cells was distinctive for each of the five major classes. Contacts on the alpha and beta cells were mainly on the dendrites in the sublamina in which a cell's major dendritic arborization was contained.

Animals↗

A systematic approach to reconstructing microcircuitry by electron microscopy of serial sections.

To observe certain quantitative features of neuronal geometry and microcircuitry, it is necessary to reconstruct neurons from electron micrographs of serial, ultra-thin sections. We describe here an approach to preparing, photographing, and analyzing moderately long series (100-500 sections). A series is prepared using an assembly line approach: one operator cuts while a second mounts ribbons of sections using various mechanical aids. Photographs are taken in the electron microscope at low magnification and high accelerating voltage. Sequential negatives are aligned using an image combiner and copied, using quasi-coherent illumination, onto 35 mm film. The resulting "movie' is mounted on a precision film transport mounted on an X-Y stage controlled by stepping motors. The movie is viewed through a high resolution video system while a video storage device and switching system permit rapid alternation between frames for comparisons. The profiles of a process in successive frames are "microaligned' by small adjustments of the transport's X-Y position. The absolute X-Y biological coordinates for each frame and the correction necessary to bring it into alignment are stored in a Z80 microprocessor as a process vector. When the movie is re-examined with the stepping motors under control of the computer, the microaligned process shows almost no frame-to-frame jitter. The process vector may be used to generate a "branch schematic' of the neuron. The microaligned profiles can also be digitized and displayed as a reconstruction using a PDP 11/34 computer. Uses of the approach are presented with examples from the cat retina and visual cortex.

Animals↗

Spatiotemporal organization of cat lateral geniculate receptive fields.

Spatial and temporal properties of LGN receptive fields were studied by flashing a small bar of light across the field in 28 discrete steps. The flashes at each of the spatial positions were used to produce 28 PST histograms. These histograms were in turn displayed as a plane, with space on the chi axis, time on the psi axis, and probability of firing on the zota axis. These response planes demonstrate that the terms on, off, center, and surround do not adequately describe when the simplest LGN receptive field. We, therefore, introduce a new terminology describing the four major spatiotemporal components of LGN fields. The primary excitatory (PE) domain corresponds to the strongest excitatory response, the secondary excitatory (SE) domain corresponds to the second-strongest excitatory domain, the primary inhibitory (PI) domain corresponds to the strongest inhibitory domain and, finally, the secondary inhibitory (SI) domain corresponds to the second-strongest inhibitory domain. Based on the arrangement of these four domains, it is possible to divide LGN fields into four major categories: 1) homogeneous-on, on-center receptive fields which have a spatially homogeneous distribution of domains; 2) homogeneous-off, off-center receptive fields which have a spatially homogeneous distribution of domains; 3) heterogeneous-on, on-center receptive fields which have a spatially heterogeneous distribution of domains; and 4) heterogeneous-off, off-center receptive fields which have a spatially heterogeneous distribution of domains; 3) heterogeneous-on, on-center receptive fields which have a spatially heterogeneous distribution of domains; and 4) heterogeneous-off, off-center receptive fields which have a spatially heterogeneous distribution of domains. Using grating, it can be demonstrated that our heterogeneous/homogeneous fields correspond to X/Y fields, respectively. These data lead us to suggest that retinal PE domains generage LGN PE and SI domains, while retinal SE domains generate LGN SE and SI domains.

Animals↗

Interactions between cat lateral geniculate neurons.

Action potentials of 31 pairs of cat LGN neurons were recorded on single electrodes. Cross-correlograms, response planes, and a new function, the logical response plane, were calculated. The cross-correlograms between these pairs revealed four interactive classes: 1) a class with a flat cross-correlogram, 2) a class with a peak in the center of the cross-correlogram seen both during spontaneous activity and during driven activity, 3) a class with an inhibitory dip in the center of the cross-correlogram seen only when the cell pairs were driven by a stimulus and predicted by the shift predictor, 4) finally, a class with an inhibitory dip in the center of the cross-correlogram seen both during driven activity and spontaneous activity and not predicted by the shift predictor. Response plane pairs calculated for the unit pairs with flat cross-correlograms showed no predictable relationship. The pairs with a dip in the center of the cross-correlograms always had response planes that were antagonistic (i.e., approximate negative images of each other). When one cell was excited, the second cell was inhibited, and vice versa. More detailed analysis, using the logical response plane, demonstrated that the majority of the antagonistic response planes had either excitatory or inhibitory overlap. That is, the cells were simultaneously excited and/or inhibited at specific spatiotemporal loci on the response plane. These data are consistent with a feed-forward inhibitory circuit in LGN. Furthermore, the data suggest that retinal centers (PE domains) produce LGN excitatory centers (PE domains) and inhibitory centers (PI domains). In turn, retinal excitatory surrounds (SE domains) produce LGN excitatory surrounds (SE domains) and inhibitory surrounds (SI domains).

Action Potentials↗

Cytoskeletal organization following cannabinoid treatment in undifferentiated and differentiated PC12 cells.

Confocal microscopy in association with three-dimensional reconstruction was used to examine the changes in the microtubules and microfilaments following cannabinoid treatment of PC12 cells. Microtubules and microfilaments were disrupted in a dose-dependent manner following treatment with 10-30 microM delta 9-tetrahydrocannabinol (THC). A disruption of microtubules and microfilaments was observed following treatment with 30 microM cannabidiol and cannabinol. The amount of microtubules and microfilaments was reduced in a dose-dependent manner following treatment with 10 and 20 microM THC. Cannabidiol and cannabinol reduced the amount of microtubules and microfilaments; however, the reduction was less than that observed with THC treatment. Following the addition of nerve growth factor, differentiated PC12 cells were generally more sensitive to cannabinoid treatments than undifferentiated cells. The possible mechanisms that may account for the changes in microtubules and microfilaments following cannabinoid treatment are discussed.

Actin Cytoskeleton↗