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Biomedical subjects

D I Vaney

Publications and source records attributed to D I Vaney.

At least 37 records · Page 2Linked to original sources

Rod-signal interneurons in the rabbit retina: 1. Rod bipolar cells.

The cellular morphology and topographic distribution of the rod bipolar cells in the rabbit retina have been investigated by selective labelling with protein kinase C-immunohistochemistry (Negishi et al., Neurosci, Lett. 94:247-252, 1988) and by Lucifer Yellow injection of microscopically identified cells in a superfused retinal preparation. The distribution of the rod bipolar cells parallels that of their input neurons, the rod photoreceptors, in that the rod bipolars reach maximum densities of 5,000-7,000 cells/mm2 on the inferior and superior flanks of the visual streak, dropping to slightly lower densities at the peak visual streak. The centre-to-periphery density gradient of the rod bipolars is about 2.5:1, and the density ratio of rods to rod bipolars shows little variation across the retina, ranging from 43:1 in superior retina to 58:1 in inferior retina. The dendritic field area of the rod bipolar cells increases from 600 microns2 on the visual streak to 1,200 microns2 in the far-superior retina, with each point on the retina overlapped by 2.5-3.5 dendritic fields. The axonal field area of the rod bipolar cells increases from about 100 microns2 at the peak visual streak to about 250 microns2 at the retina edge, and the axonal field coverage ranges from 0.55 in the visual streak to about 0.8 in peripheral retina. Although there appear to be gaps in the local array of rod bipolar somata, these areas are covered by the axonal arbours of neighbouring rod bipolar cells.

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Rod-signal interneurons in the rabbit retina: 2. AII amacrine cells.

AII amacrine cells, which are the third-order neurons in the rod pathway, can be differentially labelled in rabbit retina by injecting Nuclear Yellow into the posterior chamber. Under ultraviolet excitation, the labelled retina appears strongly metachromatic, with the AII nuclei fluorescing silvery-yellow and the nuclei of other amacrine cells fluorescing blue. Labelled AII cells were injected with Lucifer Yellow under direct microscopic control in a superfused retinal preparation, and the dye was later photoconverted to an opaque reaction product. Rabbit AII amacrines, which number about 525,000 cells, reach a maximum density of 2,500-3,000 cells/mm2 on the peak visual streak, dropping to 400-500 cells/mm2 at the superior margin. These narrow-field amacrines have a bistratified dendritic morphology, with distinctive "lobular appendages" in sublamina a of the inner plexiform layer and wider ranging "arboreal dendrites" in sublamina b. Although the lobular field area increases 10-fold from the visual streak to the far periphery, the lobular field coverage is almost uniform across the retina, averaging 1.0 in inferior retina and 0.8 in superior retina. The dendritic field area of the arboreal dendrites also increases with eccentricity from the visual streak, but there are pronounced differences between inferior and superior retina. The arboreal fields are 2 to 3 times larger than the lobular fields throughout the inferior retina but up to 15 times larger in the superior retina. The arboreal field overlap is only 1.8 at the peak visual streak, increasing slightly to about 2.4 over most of the inferior retina; the overlap increases sharply in the superior retina, however, reaching values of 10 or more in the far periphery. Both the lobular and arboreal fields of AII cells are spaced more regularly than the somata, thus covering apparent gaps in the somatic array. An analysis of the potential convergence and divergence between rod bipolar cells and AII amacrine cells in the rabbit retina indicates that the neuronal architecture of the rod circuit is not organized in a uniform module that is simply scaled-up from central to peripheral retina. Moreover, peripheral fields in the superior and inferior retina that have equivalent densities of interneurons show markedly different rod bipolar----AII amacrine convergence ratios, with the result that many more rod photoreceptors converge on an AII amacrine cell in the superior retina than in the inferior retina.(ABSTRACT TRUNCATED AT 400 WORDS)

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Many diverse types of retinal neurons show tracer coupling when injected with biocytin or Neurobiotin.

This study demonstrates that the junctional connections between rod-signal interneurons in mammalian retina can be visualized by tracer coupling, following intracellular injection of the biotinylated compounds, biocytin and Neurobiotin. In addition, many other types of retinal neurons -including B-type horizontal cells and several types of retinal ganglion cells-show specific patterns of tracer coupling, usually to cells of the same neuronal type but occasionally to cells of other neuronal classes. These findings suggest that electronic transmission occurs commonly throughout the retina and, consequently, diverse types of retinal neurons may form functional networks of coupled cells.

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The retinae of Prototherian mammals possess neuronal types that are characteristic of non-mammalian retinae.

This study has shown that the retinae of Prototherian (egg-laying) mammals possess two neuronal types that are present in non-mammalian retinae, but absent or morphologically different in the retinae of Eutherian (placental) mammals. First, endogenous serotonin-like immunoreactivity has been localized in a population of presumptive amacrine cells in the platypus retina, the first such report in a mammalian retina. Second, the protein kinase C-immunoreactive (PKC-IR) bipolar cells in the echidna retina appear similar to the PKC-IR bipolars in the chicken retina, in that their dendrites give rise to a Landolt's club and their axons are multistratified. By contrast, the PKC-IR rod bipolar cells in the rabbit and in the brushtail possum, a Metatherian (marsupial) mammal, have no Landolt's clubs and their axons form terminal lobes in the innermost stratum of the inner plexiform layer.

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The morphology and topographic distribution of substance-P-like immunoreactive amacrine cells in the cat retina.

In cat retinal wholemounts, substance-P-like immunoreactivity (SP-IR) was localized in a distinct population of amacrines whose cell bodies were normally placed in the ganglion cell layer. Although displaced amacrines accounted for 80-95% of the SP-IR amacrines in peripheral retina, this proportion decreased considerably within the area centralis, accounting for 50-80% of the labelled cells at maximum density. The SP-IR cells in both the inner nuclear and ganglion cell layers gave rise to well-defined varicose dendrites of uniform appearance that stratified around 60% depth (S3/S4) of the inner plexiform layer. In addition, sparse fine dendrites in stratum 1 (S1) could sometimes be traced to inner nuclear cells and occasionally to displaced amacrines. The combined SP-IR cell density ranged from less than 50 cells mm-2 in the far periphery to more than 500 cells mm-2 in the area centralis; the maximum density showed little individual variation despite wide differences in the proportion of displaced cells. The 39,000 SP-IR amacrines in a mapped retina had a triangular topographic distribution, with intermediate isodensity lines extending vertically in superior retina and horizontally along both arms of the visual streak. Colocalization experiments established that all SP-IR cells in cat retina showed GABA-like immunoreactivity, and that the SP-IR amacrines were quite distinct from the cholinergic amacrines identified by choline acetyltransferase immunohistochemistry.

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Neurofibrillar long-range amacrine cells in mammalian retinae.

A distinct population of wide-field, unistratified amacrine cells are shown to be selectively stained by using neurofibrillar methods in rabbit and cat retinae. Their cell bodies may be located in the inner nuclear, inner plexiform or ganglion cell layers and they branch predominantly in stratum 2 of the inner plexiform layer. Characteristically, each cell has two or more long-range distal processes which extend for 2-3 mm beyond a more symmetrical, proximal dendritic field of 0.6-0.8 mm diameter. Although the neurofibrillar long-range amacrines account for less than 1 amacrine in 500, they achieve effective coverage of the retina by both the proximal and distal dendrites.

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GABA-like immunoreactivity in NADPH-diaphorase amacrine cells of the rabbit retina.

NADPH-diaphorase histochemistry selectively stains discrete populations of retinal interneurons in diverse mammals, including two amacrine types in the rabbit retina. In this study, we have demonstrated that most of these neurons show GABA-like immunoreactivity by combining indirect immunofluorescence and diaphorase histochemistry on frozen retinal sections. The NADPH-diaphorase amacrines account for only a small proportion of the GABA-positive cells in rabbit retina, thus reinforcing the emerging consensus that GABAergic amacrines are remarkably diverse in their morphology and function.

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GABA-like immunoreactivity in cholinergic amacrine cells of the rabbit retina.

In the ganglion cell layer of the rabbit retina, the inhibitory transmitter gamma-aminobutyric acid (GABA) and its analogues are accumulated by neurons that appear to match in size and number the population of displaced amacrine cells that synthesize the excitatory transmitter acetylcholine. In this double-label study, we have established directly that the cholinergic amacrine cells, selectively stained with diamidino-phenylindole, are strongly immunoreactive with GABA antisera. The coexistence of two classical transmitters, one excitatory and the other inhibitory, in this defined neuronal population, suggests that stimulation of the cholinergic amacrines may give rise to complex responses in their target neurons.

Acetylcholine↗

Morphological identification of serotonin-accumulating neurons in the living retina.

Neurons that accumulate the transmitter serotonin have been identified in the living retina by being labeled with 5,7-dihydroxytryptamine (5,7-HT), an autofluorescent serotonin analog. Iontophoretic injection of Lucifer yellow into the labeled cells under microscopic control revealed that the serotonin-accumulating neurons in rabbit retina constitute two morphological types of amacrine cells, termed S1 and S2, whose distal dendrites are stratified at the inner margin of the inner plexiform layer. The dendritic overlap of the S1 type is extraordinarily large: each point on the retina is covered by the fields of 550 to 900 S1 amacrines, and 6 to 8 meters of their dendrites are packed into each square millimeter of retina. Such a pervasive neuropil may provide an effective substrate for diffuse transmitter release, as proposed for serotonergic fibers elsewhere in the central nervous system.

5,7-Dihydroxytryptamine↗

The morphology and topographic distribution of AII amacrine cells in the cat retina.

When cat retina is incubated in vitro with the fluorescent dye, 4',6-diamidino-2-phenyl-indole (DAPI), a uniform population of neurons is brightly labelled at the inner border of the inner nuclear layer. The dendritic morphology of the DAPI-labelled cells was defined by iontophoretic injection of Lucifer yellow under direct microscopic control: all the filled cells had the narrow-field bistratified morphology that is distinctive of the AII amacrine cells previously described from Golgi-stained retinae. Although the AII amacrines are principal interneurons in the rod-signal pathway, their density distribution does not follow the topography of the rod receptors, but peaks in the central area like the cone receptors and the ganglion cells. There are some 512 000 AII amacrines in the cat retina and their density ranges from 500 cells per square millimetre at the superior margin to 5300 cells per square millimetre in the centre (retinal area is 450 mm2). The isodensity contours are kite-shaped, particularly at intermediate densities, with a horizontal elongation towards nasal retina. The cell body size and the dendritic dimensions of AII amacrines increase with decreasing cell density. The lobular dendrites in sublamina a of the inner plexiform layer span a restricted field of 16-45 microns diameter, while the arboreal dendrites in sublamina b form a varicose tree of 18-95 microns diameter. The dendritic field coverage of the lobular appendages is close to 1.0 (+/- 0.2) at all eccentricities whereas the coverage of the arboreal dendrites doubles within the first 1.5 mm and then remains constant at 3.8 (+/- 0.7) throughout the periphery.

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'Coronate' amacrine cells in the rabbit retina have the 'starburst' dendritic morphology.

Acetylcholine-synthesizing cells in the rabbit retina are symmetrically distributed about the inner plexiform layer: one population of cholinergic amacrines has cell bodies in the inner nuclear layer and an equivalent population of displaced amacrines has cell bodies in the ganglion cell layer. It has been suggested that the morphological correlates of the acetylcholine-synthesizing cells are either coronate amacrine cells or starburst amacrine cells. Coronate cells have a characteristic nuclear morphology and can be selectively labelled by neurofibrillar methods or with the fluorescent dye4',6-diamidino-2-phenyl-indole (DAPI). Starburst cells have a characteristic dendritic morphology but have only been described from Golgi-stained retinae. This paper bridges the gap between the previous studies. DAPI-labelled coronate cells were impaled with a micropipette under microscopic control and filled with Lucifer yellow by iontophoresis. The results show that the coronate amacrines in the ganglion cell layer are type b starburst cells, and that those DAPI-labelled neurones in the inner nuclear layer with a coronate-like nuclear morphology are type a starburst cells. At a given eccentricity the dendritic field diameter of type a starburst cells is about 1.13 times larger than that of type b starburst cells. The dendritic field coverage of coronate (type b starburst) cells increases linearly with decreasing coronate cell density and ranges from 25 on the peak visual streak to 70+ in the superior periphery.

Acetylcholine↗

Single unit receptive fields in rabbit primary binocular cortex.

The receptive fields of 125 single units recorded from the binocular region of rabbit primary visual cortex have been analysed. The population of 43% radially symmetric, 23% directional, and 23% orientation selective units is similar to that of rabbit monocular visual cortex. The relative scarcity of orientation selective units and the absence of orientation columns differentiates rabbit from cat primary visual cortex. However, the majority of binocular units had similar receptive fields in each eye and very unconventional receptive fields were not encountered. Tested binocular units demonstrated summation upon simultaneous stimulation of both receptive fields. In conjunction with findings reported elsewhere, these results suggest that rabbit and cat possess a similar provision for binocular vision in spite of some differences in their cortical organisation.

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Matching populations of amacrine cells in the inner nuclear and ganglion cell layers of the rabbit retina.

In rabbit retina, neurofibrillar methods stain two populations of amacrines whose cell before are located on either side of the inner plexiform layer: one in the ganglion cell layer and the other at the inner margin of the inner nuclear layer. The stained amacrines in the ganglion cell layer have the distinctive cytology of "coronate" amacrines described from Nissl-stained retina (Vaney, '80a) and account from about 85% of the displaced amacrines in the rabbit retina. The coronate amacrines have a streak topography similar to that of the ganglion cells; they comprise about 32% of the neurons in the ganglion cells layer although their proportion increases with eccentricity from the visual streak. The cytology of the neurofibrillar stained amacrines in the inner nuclear layer resembles that of the displaced amacrines and their densities are almost equal. The cell bodies of the stained amacrines are smaller than those of the displaced amacrines but larger than most others in the inner nuclear layer; they account for some 2% of the neurons in the amacrine sublayer. Although the cell bodies of both populations are distributed rather evenly, the mosaic of the inner nuclear layer cells is more regular than that of the ganglion layer cells. We propose that the two populations of amacrines stained by neurofibrillar methods correspond to the acetylcholine synthesizing cells labelled by Masland and Mills ('79).

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Rabbit retinal ganglion cells. Receptive field classification and axonal conduction properties.

The classification of concentrically organized receptive fields of rabbit retinal ganglion cells was extended along similar lines to that in the cat by distinguishing brisk and sluggish classes and then sustained and transient types of each. Quantitative measures of responsiveness to stationary and to moving stimuli revealed characteristic features which distinguished these classes. Brisk-transient and brisk-sustained classes are not as distinct from each other as in the cat: centre size distributions overlapped almost completely and there was also substantial overlap of axonal conduction properties whether expressed in terms of latency or conduction velocity between two central stimulus sites. Representatives of every class of rabbit ganglion cells sent axons to the superior colliculus.

Action Potentials↗

A quantitative comparison between the ganglion cell populations and axonal outflows of the visual streak and periphery of the rabbit retina.

A vertical density profile of the ganglion cells 2 mm temporal of the optic nerve head in the rabbit retina has been produced by counting somata in the cresyl-violet-stained, ganglion cell layer of a flat-mounted retina. Somata classified as ganglion cells were characterized by obvious Nissl staining in an extensive cytoplasm and typically had diameters greater than 9 micrometer. The accuracy of the profile, and thus of the classification criteria, has been substantiated by electron micrographic determination of the numbers of ganglion cell axons arising within local regions of known area on the same retina. This study indicates that Vaney and Hughes' estimate ('76) of 547,100 presumed ganglion cells in the rabbit retina should be changed to 373,500 ganglion cells. The latter value is within the statistical error of their optic nerve count of 394,000 fibers. The mean diameter of ganglion cells 6 mm from the visual streak in the inferior periphery (density: 550 cells/mm2) was 28% greater than that of cells on the peak of the streak (density: 5,400 cells/mm2), although the form of the ganglion cell diameter distribution did not change markedly with eccentricity. The increase in the mean size of ganglion cells in the periphery appeared to be approximately matched by an increase in the size of their axons. Larger axons became myelinated farther from the edge of the myelinated band than did smaller axons. Within the ganglion cell layer there was another population of cells which were quite distinct from the obvious neuroglia: Their nuclei were similar to those of the larger ganglion cells and many appeared to have Nissl granules within their limited cytoplasm. About half of this heterogeneous population was classified as "coronate cells," which were characterized by the partial nuclear encapsulation of their eccentric cytoplasm.

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