PubMed Health⌕ Search

Biomedical subjects

R H Masland

Publications and source records attributed to R H Masland.

At least 19 recordsLinked to original sources
PubMed1995

The organization of the inner nuclear layer of the rabbit retina.

The initial goal of this study was to establish an accounting of the major classes of cells present in the inner nuclear layer (INL) of the rabbit's retina. Series of 80-100 radial sections 1 micron thick were cut from retinal blocks dissected at intervals along the vertical meridian. They were photographed at high magnification in the light microscope. By visualizing the initial segments of processes leaving the somata, we could identify each cell as a bipolar, amacrine, horizontal, or Müller cell. The identifications made by light microscopy were confirmed by electron microscopy of alternating ultrathin sections. On average, bipolar cells made up 41% of the total INL cells, amacrine cells 32%, horizontal cells 1.5%, and Müller cells 24%. These fractions varied relatively little across the retina or among different animals. We next immunolabeled the rod bipolar cells of whole-mounted retinas with antibodies against protein kinase C, using FITC as the visualizing agent. The same retinas were counterstained with a DNA-binding probe that fluoresces at longer wavelengths. Serial optical horizontal sections of the double-labeled wholemounts were made by confocal microscopy. On average, rod bipolars accounted for 10% of the total INL cells. By subtraction, the cone bipolars made up 31% of the total cells. We conclude that cone bipolars substantially outnumber rod bipolars, even in a retina in which rods outnumber cones by more than 20:1. Using the base of reference created here, a similar analysis can be carried out for other subclasses of retinal neuron. Because the analysis does not depend on absolute cell densities or corrections for shrinkage, data acquired by different histochemical techniques may be combined.

Animals↗
PubMed1994

Receptive fields and dendritic structure of directionally selective retinal ganglion cells.

We studied the relationship between the receptive fields of directionally selective retinal ganglion cells and the dendritic arbors of the same cells. The cells were recorded from extracellularly under visual control and then injected with Lucifer yellow. The arbor of Lucifer-filled dendrites could then be directly compared with the properties of the receptive field. A large population of on-off directionally selective cells was injected and drawn. The directionally selective ganglion cells had bistratified receptive fields similar to those previously described by others in the central retina. In the periphery, the dendritic fields became larger, rounder, and sparser than centrally. The diameters of the dendrites were measured in living or lightly fixed retinas; they were found to be somewhat larger than previously estimated by electron microscopy. The local structure of the dendritic arbor bore no obvious relation to the directional properties of the cell. The receptive fields of most cells were centered symmetrically around their dendritic fields. For about 10% of the cells, however, the receptive field was displaced. The displacement was always toward the preferred direction, relative to the dendritic field. The meaning of these shifts is not clear. In both cases, the diameter of the receptive field exceeded the diameter of the dendritic field only slightly; in our sample, the diameters of the receptive fields averaged 6% larger than the dendritic fields. This means that the neurons afferent to the directionally selective ganglion cells must either have narrow dendritic fields or, if they are wide spreading, have dendrites that do not conduct effectively along their length. It also means that the observed spread of neurobiotin between DS ganglion cells (Vaney, 1991) must be due to a very few gap junctions, or to some mechanism other than a gap junction.

Animals↗
PubMed1993

Responses of the starburst amacrine cells to moving stimuli.

1. Rabbit retinas were isolated from the eye and incubated in the presence of 3H-choline. Samples of retina taken from a defined midperipheral eccentricity were spread over the domed end of a fiberoptic bundle that formed the floor of a superfusion chamber. The rate of release of labeled acetylcholine by the starburst amacrine cells was studied. 2. When the retina was stimulated by moving gratings, the cells vigorously increased their secretion of acetylcholine. Responses were observed when the bars were as small as 60 microns in width. Systematically varying the spatial and temporal frequency of stimulation revealed that temporal frequency was the dominant variable: the cells responded best to stimuli of 1-4 Hz, whether those stimuli were flashing lights, fine gratings moving slowly, or coarse gratings moving rapidly. 3. With temporal frequency constant, the cells' responses decreased as the spatial frequency of the grating increased. The decreased response to fine gratings is most likely due, at least in part, to lateral interactions that become stronger as the light and dark bars become more closely spaced. These could occur in either the outer or inner retina. 4. The velocity tuning curve for the starburst cells' release of acetylcholine matched fairly well the velocity tuning of ON-OFF directionally selective cells in the rabbit. It did not correspond at all well with the tuning curve for the ON directionally selective cells. If the ON cells receive input from the starburst cells, that input appears to be quite indirect.(ABSTRACT TRUNCATED AT 400 WORDS)

Acceleration↗
PubMed1993

Selective accumulation of diamidino yellow and chromomycin A3 by retinal glial cells.

We applied the fluorescent DNA stains diamidino yellow (DY) and chromomycin A3 to rat and rabbit retinas in vivo and in vitro. They accumulated in the nuclei of a subpopulation of cells of the inner nuclear layer. The number and distribution of the fluorochrome-accumulating cells were similar to those of the Müller glia, and double-labeling experiments showed that the cells accumulating DY or chromomycin A3 contained oriented filaments of vimentin. The fluorochromes also accumulated in the sparse astrocytes and oligodendrocytes located among the myelinated fibers of the rabbit central retina. Specific accumulation in retinal glia occurred only when the fluorochromes were applied to living retinas. If the plasma membranes were disrupted by fixation or exposure to detergent, most retinal cells were stained. This indicates that the locus of specificity is the entry of the molecules into the cells. When applied to living retinas, other DNA stains selectively accumulate in subclasses of retinal neurons. Why DNA-binding molecules should selectively cross the membranes of either retinal neurons or retinal glia remains an unsolved problem.

Amidines↗
PubMed1993

Developmental variation in the structure of the retina.

Events traditionally called "developmental errors" are known to occur in both vertebrate and invertebrate nervous systems. This study was concerned with the frequency and mode of generation of such events in the mammalian retina. We studied three anomalous structures observed in the rabbit's retina after staining of the cell populations that accumulate indoleamines: type 3 cells, stray processes in the optic fiber layer, and displaced cells. They were counted in rabbit retinas prepared as whole-mounts, and in most cases topological maps were made. For comparison, the conventional indoleamine-accumulating amacrine cells and the tyrosine hydroxylase (TH)-positive cells, which are members of the mammalian retina's recognized complement of amacrine cells, were also counted. A further comparison was made with the number and distribution of TH-positive amacrine cells in highly inbred mice. The ordinary amacrine cells did not vary much in number from animal to animal. Especially in inbred mice, the reproducibility was striking: the extreme variation in number of TH amacrine cells between any two of the 14 retinas studied was 22%, and the mean difference between two eyes of individual mice was 2.5 +/- 1.7%. The three anomalous structures were rare and variable. Their numbers varied more than fourfold from animal to animal. However, their numbers in two eyes from the same animal varied by an overall average of only 14 +/- 10%. The anomalous structures were present in all rabbits, and their morphology was the same in all cases: they are under precise control by the developmental program. The anomalous cells share many phenotypic features with the regular amacrine cells of the indoleamine-accumulating class.(ABSTRACT TRUNCATED AT 250 WORDS)

Amines↗
PubMed1992

Direct visualization of the dendritic and receptive fields of directionally selective retinal ganglion cells.

Optical methods were used to locate the cell bodies of directionally selective ganglion cells in isolated rabbit retinas. These neurons detect the direction in which images move across the retinal surface and transmit that information to the brain. The receptive field of each identified cell was determined, after which the cell was injected with Lucifer yellow. An image of the receptive field border was then projected onto the fluorescent image of the dendrites, allowing precise comparison between them. The size of the receptive field matched closely the size of the dendritic arbor of that cell. This result restricts the types of convergence that can be postulated in modeling the mechanism of retinal directional selectivity.

Amidines↗
PubMed1992

Co-release of acetylcholine and GABA by the starburst amacrine cells.

Rabbit retinas were isolated from the eye and maintained in vitro. When they were incubated for 60 min in the presence of 3H-GABA, subsequent autoradiography showed radioactivity to be present primarily in amacrine cells. Under these conditions, most of the radioactivity contained in the retinas remained in the chemical form of GABA. Autoradiography and immunohistochemistry of alternate sections showed the amacrine cells that accumulate 3H-GABA to be the same cells that contain endogenous GABA immunoreactivity. These include the starburst cells, the indoleamine-accumulating cells, and other, as yet unidentified amacrine cells. The localization confirms previous immunohistochemical findings. When retinas containing 3H-GABA were expressed to elevated concentrations of K+, their content of 3H-GABA decreased. Autoradiography showed a reduced 3H-GABA content in all of the cells that contained 3H-GABA. Since those include the starburst cells, previously shown to be cholinergic, the finding demonstrates that the starburst cells release both ACh and GABA. Retinas simultaneously labeled with 14C-GABA and 3H-ACh were superfused, and the release of radioactive compounds from the retina was studied. Depolarization by elevated K+ caused an increased recovery of both ACh and GABA in the superfusate, but the predominant mechanisms of their release appeared to be different. The stimulated release of ACh was entirely Ca2+ dependent, while the release of radioactivity originating from GABA was much less so. A concentration-dependent counterflux (homoexchange) of intracellular GABA was demonstrated by raising the extracellular concentration of GABA (or nipecotic acid). These results suggest that a large outward flux of GABA occurs via the GABA transporter, probably by the potential-sensitive mechanism studied by Schwartz (1982, 1987). Stimulation of double-labeled retinas by flashing light or moving bars always increased the release of ACh, and the release was entirely dependent on the presence of extracellular Ca2+. Stimulation with light never caused a detectable release of GABA. This was unexpected, since the two neurotransmitters are present in the same amacrine cells: stimulation adequate to release one neurotransmitter should release both.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholine↗
PubMed1989

Connections of indoleamine-accumulating cells in the rabbit retina.

To study the connections of the neurons of the rabbit retina that accumulate indoleamines, we injected 5,7-dihydroxytryptamine into the vitreous body. It accumulated within a subset of amacrine cells and could be visualized there by aldehyde-induced fluorescence. The fluorescent labeling was photo-converted to an insoluble, osmiophilic product by irradiation in the presence of diaminobenzidine, and the tissue was examined by electron microscopy. Preservation of the structure of the tissue after photoconversion was satisfactory and the dendrites of the indoleamine-accumulating cells could easily be identified. They form a dense plexus near the junction of the inner plexiform and ganglion cell layers, where they exhibit large synaptic endings that occupy a substantial fraction of the surface of rod bipolar terminals. The dendrites of the indoleamine-accumulating cells receive input from rod bipolars at dyad synapses, where the other postsynaptic partner is a dendrite of a narrow-field, bistratified amacrine cell; in addition, they receive amacrine cell input throughout the inner plexiform layer. The only outputs we observed are reciprocal synapses onto the rod bipolar endings. Thus, these amacrine cells appear to exert an important effect on the transmission of scotopic information through the retina.

5,7-Dihydroxytryptamine↗
PubMed1989

Shape and distribution of an unusual retinal neuron.

Rabbit retinas were exposed to exogenous indoleamines and fixed with mixed aldehydes. The indoleamines were accumulated by two types of amacrine cell and by an unusual cell (type 3) that branches widely in both plexiform layers. The type 3 cells were studied after immunohistochemistry, photooxidation of the fluorescent label, or injection with Lucifer Yellow. Their cell bodies are located at the scleral margin of the inner nuclear layer. The cells' arbors in the outer plexiform layer range from 800 to 1,500 microns in diameter. A descending process crosses the inner nuclear layer and branches in layer 5 of the inner plexiform layer. The arbor in the inner retina can exceed 500 microns in diameter. The distribution of type 3 cells was mapped in a series of retinal whole mounts. The number of type 3 cells ranged from 58 to 270 in different retinas. In two retinas from a single animal, however, it was virtually identical. Type 3 cells are concentrated in the region ventral to the visual streak, so that large areas of the retina are not covered by any type 3 cell. Because of their incomplete retinal coverage and variable number from animal to animal, the type 3 cells appear to be developmental anomalies. Paradoxically, their generation must be precisely controlled because of the numerical symmetry between an individual animal's two eyes.

Aldehydes↗
PubMed1989

Indoleamine accumulation by retinal neurons exposed to blood.

Exposing rabbit retinas for one minute to an incubation medium containing 10 microliters of blood diluted in 20 ml of medium was sufficient to produce serotonin-like immunoreactivity in some of the retinal indoleamine-accumulating neurons. Retinas from rabbits that had been perfused before the eyes were removed had no detectable immunoreactivity. Our results support the conjecture that the serotonin sometimes detected in the retina originates in the blood. Why the cells have a carrier for a molecule that they do not normally contain remains unclear.

Amines↗
PubMed1989

Co-release of acetylcholine and gamma-aminobutyric acid by a retinal neuron.

Rabbit retinas were vitally stained with 4',6-diamidino-2-phenylindole (DAPI), a fluorescent compound that selectively accumulates within the cholinergic amacrine cells. The retinas were then incubated in vitro in the presence of radioactive gamma-aminobutyric acid (GABA) and autoradiographed. The cells that accumulated DAPI were found to accumulate GABA, confirming immunohistochemical evidence that the cholinergic amacrine cells contain GABA. Incubation of retinas in the presence of elevated concentrations of K+ caused them to release acetylcholine and GABA, and autoradiography showed depletion of radioactive GABA from the cholinergic amacrine cells. This indicates that the cholinergic amacrine cells can secrete acetylcholine and GABA. Retinas were double-labeled with [14C]GABA and [3H]acetylcholine, allowing simultaneous measurement of their release. The release of [14C]GABA was found to be independent of extracellular Ca2+. Radioactive GABA synthesized endogenously from [14C]glutamate behaved the same way as radioactive GABA accumulated from the medium. In the same experiments the simultaneously measured release of [3H]acetylcholine was strongly Ca2+-dependent, indicating that the releases of acetylcholine and GABA are controlled by different mechanisms. Synaptic vesicles immunologically isolated from double-labeled retinas contained much [3H]acetylcholine and little or no [14C]GABA. These results suggest that the cholinergic amacrine cells release acetylcholine primarily by vesicle exocytosis and release GABA primarily by means of a carrier.

Acetylcholine↗
PubMed1988

Amacrine cells.

Explore the source record for details and available documents.

Animals↗
PubMed1988

Photoconversion of some fluorescent markers to a diaminobenzidine product.

Retinal whole mounts, brain sections, and astrocyte cultures were labeled with various fluorescent markers. Tissues or cells were then irradiated by light in the presence of diaminobenzidine. Irradiation initiated a reaction in which specific fluorescent labeling was replaced by an insoluble diaminobenzidine product. The diaminobenzidine product is more stable than the original fluorescent labeling and can be processed for electron microscopy. In some cases, the reaction product reveals cellular detail that cannot be resolved in the fluorescent labeling. The 10 fluorescent markers tested have widely differing structures, span a broad range of wavelengths, and label several different cellular elements. The photoconversion reaction was successful with all markers and tissues tested.

Animals↗
PubMed1987

The resting release of acetylcholine by a retinal neuron.

The cholinergic amacrine cells of the rabbit retina secrete acetylcholine by two mechanisms. One is activated by stimulation of the retina by light or depolarization of the amacrine cells by K+ ions. It requires the presence of extracellular Ca2+. The second is independent of extracellular Ca2+ and is unaffected by large depolarizations of the cells. It bears some similarity to the acetylcholine 'leakage' described at the neuromuscular junction. Although the Ca2+-independent mechanism accounts for about two thirds of the total acetylcholine release in the dark, the amount of acetylcholine released in this way is small compared with the release of acetylcholine triggered by stimulation of the retina with light. Its biological significance is unclear.

Acetylcholine↗
PubMed1986

A system of indoleamine-accumulating neurons in the rabbit retina.

The indoleamine-accumulating neurons of the rabbit retina were labeled by intraocular injection of 5,7-dihydroxytryptamine (5,7-DHT). The retinas were fixed with 2.5% paraformaldehyde and 0.2% glutaraldehyde and inspected by fluorescence microscopy. Five kinds of cell accumulated the indoleamine. They were labeled to essentially the same brightness and remained so despite variations in the concentration at which 5,7-DHT had been applied or the duration of its application. Experiments in which 5,7-DHT was applied to retinas incubated in vitro gave identical results. To see the whole shape of the cells, we visually guided micropipettes to the fluorescent cell bodies and injected the cells with Lucifer yellow CH. To study the cells as a population, we used a new method in which the fluorescence of 5,7-DHT is photochemically converted to an insoluble diaminobenzidine product. The dendrites of all of the indoleamine-accumulating cells were then simultaneously visible. Used together, these techniques revealed an interrelated system of indoleamine-accumulating neurons. All of the cells contribute processes to a dendritic plexus that lies at the inner margin of the inner plexiform layer. The plexus is roughly 4 micron thick. It is pierced by the stalks of the Müller cells and is occasionally interrupted by ganglion cell bodies, where they extend above the average margin of the ganglion cell layer. Otherwise it fills much of the space at the junction of the plexiform and ganglion cell layers. The type 1 and type 2 cells are amacrine cells with cell bodies at the inner margin of the inner nuclear layer. They have 5-8 radially branching primary dendrites which extend horizontally across the inner plexiform layer before descending to join the dendritic plexus. They differ from each other in cell body shape, dendritic morphology, and the course of their dendrites within the inner plexiform layer. Each has a "displaced" counterpart, with a morphology similar to the type 1 or type 2 cell but with a cell body located in the ganglion cell layer. The displaced cells are separate functional elements because, in contrast to the type 1 and type 2 cells, they have no dendrites (and hence can have no synaptic connections) in the outer part of the inner plexiform layer. The fifth kind of cell (type 3) appears not to have been described before. Its cell body is located at the outer margin of the inner nuclear layer.(ABSTRACT TRUNCATED AT 400 WORDS)

5,7-Dihydroxytryptamine↗
PubMed1985

Local order among the dendrites of an amacrine cell population.

The cholinergic amacrine cells of the rabbit retina branch within a narrow stratum of the retina's inner synaptic layer, and their dendritic fields overlap as much as 70-fold. Because each cell's dendrites have many branches, the overlap must create a dense meshwork of cholinergic dendrites. To learn how the overlapping dendrites are positioned with respect to each other, we filled the dendrites of groups of neighboring cells with Lucifer Yellow CH. The cholinergic amacrine cells were selectively stained by intraocular injection of the fluorescent molecule 4,6-diamidino-2-phenylindole. The retinas were then fixed with 2% paraformaldehyde and 0.01% glutaraldehyde. The stained cells were penetrated under visual control by Lucifer Yellow-filled micropipettes. A systematic arrangement of the dendrites was observed. When a pair of cells was injected, their dendrites were often seen to lie alongside each other. In the terminal dendritic region, there are virtually no dendrites that do not end in apposition to a dendrite of a neighboring cholinergic amacrine cell. When small clusters of nearby cells were injected, an ordered microstructure appeared. The dendrites of the cells join together to form curving bundles, which enclose spaces that rarely contain any cholinergic dendrites: the appearance of the dendritic mosaic is that of a lattice with a repeating unit roughly 10 microns in diameter. The significance of this ordering is not certain, but it is possible that the repeating structural unit participates in a modular functional arrangement.

Animals↗