"Small-tufted" ganglion cells and two visual systems for the detection of object motion in rabbit retina.

Small-tufted (ST) ganglion cells of rabbit retina are divided into eight types based upon morphology, branching pattern, level of dendritic stratification, and quantitative dimensional analysis. Only one of these types has been previously characterized in Golgi preparations, and four may be discerned in the work of others. Given their small dendritic-field size, and assuming uniform mosaics of each across the retina, ST cells comprise about 45% of all rabbit ganglion cells, and are therefore of major functional significance. Four ST cells occur as two paramorphic (a/b) pairs, and thus belong to class III, as previously defined. Four branch in sublaminae a and b of the inner plexiform layer (IPL) and therefore belong to class IV. ST cells have small cell bodies 10-15 microm in diameter, small axons 0.7-1.3 microm in diameter, and small dendritic-field diameters, 40-110 microm in mid-visual streak. The dendrites of ST cells are highly branched, and bear spines and appendages of varying length, but vary from type to type. Class III.2 cells and class III.3 cells are partly bistratified. Class IV small-tufted cells differ characteristically in multiple features of dendritic branching and stratification. Class III small-tufted cells apparently have concentric (ON-center and OFF-center) receptive fields and may have "sluggish-transient" (class III.2) and "sluggish-sustained" (class III.3) physiology. Class IV cells include the "local-edge-detector" (LED) (class IVst1), and are all expected to give ON-OFF responses to small, centered, slowly moving visual stimuli. Based upon systematic variation in dendritic-field size across the retina, ST cells may be divided into two groups. In this "universal prey" species, they may belong to two systems of motion detection, typified by ON-OFF directionally selective and LED ganglion cells, respectively, specialized for detection of rapid motion at the horizon for land-based predators, and slow motion for airborne predators.

A structural basis for omnidirectional connections between starburst amacrine cells and directionally selective ganglion cells in rabbit retina, with associated bipolar cells.

Directionally selective (DS) ganglion cells of rabbit retina are of two principal types. ON DS ganglion cells prefer low velocity in one of three directions of movement and project axons to the accessory optic system (AOS), whereas ON-OFF DS ganglion cells prefer higher velocity in one of four directions and project to tectum and thalamus. Each has a distinct, recognizable dendritic morphology, based upon the correlation of form, physiology, and central projections. In previous Golgi studies, ON and ON-OFF DS cells were found to be partly co-stratified, and ON-OFF DS cells were found to co-stratify with starburst amacrine (SA) cells, the cholinergic amacrine cells of the retina, which also contain elevated levels of GABA. SA cells are radially symmetrical, have synaptic boutons in a distal annular zone of its dendritic tree, are presynaptic primarily to ganglion cell dendrites, co-stratify with ON-OFF DS ganglion cells, and contain the neurotransmitters shown pharmacologically to be involved in DS responses. For these reasons, SA cells are thought to play a role in the DS mechanism. Several models of this mechanism have utilized SA cell dendritic geometry in a centrifugal, radial format to impose directional inputs on DS ganglion cells. The opportunity to examine Golgi preparations containing ON DS ganglion cells that exhibit dendritic field overlap with both starburst amacrine cells and ON-OFF DS ganglion cells has resulted in several new findings. Co-stratification of ON DS ganglion cells and SA cells was demonstrated directly. Secondly, the boutons of single starburst amacrine cells make close contact in different lamellae of the starburst substratum in sublamina b of the inner plexiform layer (IPL) with three adjacent ON-OFF DS ganglion cells, which because of their considerable dendritic-field overlap must prefer different directions of motion. Thirdly, nearby presynaptic boutons of single SA cells make close contact with both ON and ON-OFF directionally selective ganglion cells. Single SA cells thus traverse all the lamellae of the starburst/cholinergic substratum. Fourthly, no directional bias is shown by vectors connecting the origins of dendritic sectors and distal synaptic boutons of starburst amacrine cells in those sectors that are in close contact with the dendrites of single ON or ON-OFF directionally selective ganglion cells. Fifthly, at least two distinct types of cone bipolar cell, nb1 and nb2, participate in the neural circuitry of directional selectivity for ON and ON-OFF DS ganglion cells, and nb1 cells co-stratify with ON DS cells. As a consequence of the second, third, and fourth points, starburst amacrine cells appear to be indiscriminate in their connections with DS ganglion cells, and therefore are unlikely to be the primary conduits for directionally selective information to retinal ganglion cells. This result is consistent with pharmacological studies showing that cholinergic antagonists do not block directional selectivity and a study showing that laser-ablation of SA cells does not reduce the directional selectivity of overlapping ON-OFF DS ganglion cells.

The zeta cell: a new ganglion cell type in cat retina.

We define a new morphological type of ganglion cell in cat retina by using intracellular staining in vitro. The zeta cell has a small soma, slender axon, and compact, tufted, unistratified dendritic arbor. Dendritic fields were intermediate in size among cat ganglion cells, typically twice the diameter of beta cell fields. They were smallest in the nasal visual streak (<280 microm diameter), especially near the area centralis (60-150 microm diameter), and largest in the nonstreak periphery (maximum diameter 570 microm). Fields sizes were symmetric about the nasotemporal raphe except near the visual streak, where nasal fields were smaller than temporal ones. Zeta-cell dendrites ramified near the boundary between sublaminae a and b (OFF and ON sublayers) of the inner plexiform layer, occupying the narrow gap separating the dendrites of ON and OFF alpha cells. There was no evidence for separate ON and OFF types of zeta cell. Retrograde labeling studies revealed that both nasally and temporally located zeta cells project to the contralateral superior colliculus, whereas few project to the ipsilateral colliculus or to any subdivision of the dorsal lateral geniculate nucleus. The zeta cell's morphology and projection patterns suggest that it corresponds to the ON-OFF phasic W-cell (also known as the local edge detector) of physiological studies. Zeta cells have particularly small dendritic fields in the visual streak, presumably because they are disproportionately represented in the streak in comparison with other ganglion cell types. These conditions are consistent with optimal spatial resolution along the retinal projection of the visual horizon rather than principally at the center of gaze. Strong commonalities with similar ganglion cell types in ferret, rabbit, and monkey suggest that "zeta-like" cells may be a universal feature of the mammalian retina.

Regional topography of rod and immunocytochemically characterized "blue" and "green" cone photoreceptors in rabbit retina.

Evidence from several sources indicates that the photoreceptors of rabbit retina include rods, green cones and blue cones, and that blue-green color opponency occurs in select retinal ganglion cells. One of us (Famiglietti) has identified wide-field cone bipolar cells as probable blue-cone-selective bipolars, and type C horizontal cells as possibly connected to blue cones. We wished to extend the analysis of blue cone pathways in rabbit retina and to characterize the topographic distribution of blue and green cones. Two monoclonal antibodies raised against chicken visual pigments are reported to label medium- and long-wavelength cones (COS-1) and short-wavelength cones (OS-2) in all mammalian retinas studied thus far (Szél and colleagues). Using selective labeling with these two antibodies and a nonselective method in nasal and temporal halves of the same retinas, we have found that densities of photoreceptors vary systematically, depending upon the size of the eye and age of the animal. In 'standard' New Zealand rabbits of 2-3 kg (2-3 months old), rods reached a peak density of about 300,000/mm2 just dorsal to the visual streak, while cones exhibit peak density at mid-visual streak of about 18,000/mm2. Published measurements of visual acuity in rabbit are less than predicted by this calculation. The ratio of cones to rods is significantly higher in ventral retina, where the density of cones declines to a plateau of 10,000-12,000/mm2, when compared to dorsal retina, where cones are uniformly distributed at a density of about 7000/mm2. The density of OS-2 labeled (presumably "blue") cones is uniformly low, 1000-1500/mm2, in a wide expanse that includes dorsal retina, the visual streak, and much of ventral retina, except for a region of higher density along the vertical midline. We confirm that there is a far ventral horizontal region near the perimeter that is populated exclusively by a high density (about 13,000/mm2) of OS-2-positive cones (Juliusson and colleagues). This region does not extend to the ventral retinal margin, however, where cone density drops precipitously. Transitional zones between COS-1 and OS-2 labeling, in a region of relatively high and uniform cone density, where sums of COS-1 and OS-2 labeling are higher than expected and in which weakly and strongly labeled cones are intermixed, raise questions about the identities of the visual pigment epitopes, the possibility of double labeling, and therefore the possibility of dual expression of pigments in single cones. The "inverted-T-shaped" topography of higher density OS-2 labeling raises doubts about the significance of a ventral concentration of blue cones for visual function in rabbit retina.

New metrics for analysis of dendritic branching patterns demonstrating similarities and differences in ON and ON-OFF directionally selective retinal ganglion cells.

The morphology and dendritic branching patterns of retinal ganglion cells have been studied in Golgi-impregnated, whole-mount preparations of rabbit retina. Among a large number of morphological types identified, two have been found that correspond to the morphology of ON and ON-OFF directionally selective (DS) ganglion cells identified in other studies. These two kinds of DS ganglion cell are compared with each other, as well as with examples of class I, class II, and class III cells, defined here with reference to our previous studies. Cell body, dendritic field size and branching pattern are analyzed in this paper and levels of dendritic stratification are examined in the following paper. ON DS ganglion cells are about 10% larger in soma size and about 5 times the dendritic field area of ON-OFF DS ganglion cells, when compared at the same retinal location. These two morphological types of ganglion cell can be said to define the upper and lower bounds of an intermediate range of cell body and dendritic field sizes within the whole population of ganglion cells. Nevertheless, in previous physiological studies receptive field sizes of the two types were shown to be similar. This discrepancy between morphological and physiological evidence is considered in the Discussion in terms of a model of the excitatory receptive field of ON-OFF DS ganglion cells incorporating starburst amacrine cells. A new set of metrics is introduced here for the quantitative analysis and characterization of the branching pattern of neuronal arborizations. This method compares the lengths of terminal and preterminal dendritic branches (treated separately), as a function of the distances of their origins from the soma, viewed graphically in a two-dimensional scatter plot. These values are derived from computer-aided 3D logging of the dendritic trees, and distance from the soma is measured as the shortest distance tracked along the dendritic branches. From these metrics of the "branch length distributions," scale-independent branching statistics are derived. These make use of mean branch lengths and distances, slopes of lines fitted to the distributions, and elliptical indices of scatter in the distributions. By these measures, ON and ON-OFF DS ganglion cells have similar branching patterns, which they share to varying degrees with functionally unrelated class III.1 ganglion cells. The scale of the branching patterns of ON and ON-OFF DS cells and their degree of uniformity are different, however.(ABSTRACT TRUNCATED AT 400 WORDS)

Dendritic co-stratification of ON and ON-OFF directionally selective ganglion cells with starburst amacrine cells in rabbit retina.

The morphology, dendritic branching patterns, and dendritic stratification of retinal ganglion cells have been studied in Golgi-impregnated, whole-mount preparations of rabbit retina. Among a large number of morphological types identified, two have been found that correspond to the morphology of ON and ON-OFF directionally selective (DS) ganglion cells identified in other studies. These cells have been characterized in the preceding paper in terms of their cell body size, dendritic field size, and branching pattern. In this paper, the two kinds of DS ganglion cell are compared in terms of their levels of dendritic stratification. They are compared with each other and also with examples of class III.1 cells, defined in the preceding paper with reference to our previous studies. Studies employing computer-aided, 3D reconstruction of dendritic trees, as well as analysis of a pair of ON DS and ON-OFF DS ganglion cells with overlapping dendritic trees show that the two types of DS ganglion cell partly co-stratify in the middle of sublamina b (stratum 4). The report that some ON DS ganglion cells extend a few dendrites into sublamina a is confirmed. The study of pairs of ON-OFF DS ganglion cells and starburst amacrine cells with overlapping dendritic trees reveals a precise co-stratification of these two cell types, and many points of close apposition of starburst boutons with ON-OFF DS ganglion cell dendrites in both sublaminae of the inner plexiform layer (IPL). This is confirmed by high-resolution light microscopy and by electron microscopy. It is possible to conclude, therefore, that ON DS are also partly co-stratified with type b starburst (cholinergic) amacrine cells, and are apparently also partly co-stratified with type a starburst amacrine cells, when occasional dendrites rise to that level. The co-stratification of the two kinds of DS ganglion cell is consistent with the sharing of some inputs in common, including some cone bipolar cell inputs. The co-stratification of both with starburst amacrine cells agrees with the physiological demonstration of the powerful pharmacological effects upon ON and ON-OFF DS ganglion cells reported for cholinergic agonists. The major difference in the dendritic stratification of bistratified ON-OFF DS ganglion cells and generally unistratified ON DS ganglion cells is consistent with the bisublaminar organization of ON and OFF pathways in the IPL. The problem of occasional branches of ON DS cells in sublamina a is discussed in terms of a threshold for OFF responses.(ABSTRACT TRUNCATED AT 400 WORDS)

Polyaxonal amacrine cells of rabbit retina: morphology and stratification of PA1 cells.

Polyaxonal amacrine cells are a new class of amacrine cell bearing one to six branching, axon-like processes, closely resembling the axons of Golgi type II cells found elsewhere in the central nervous system. Of the four types of polyaxonal amacrine cell that we have recognized in rabbit retina, three have been described previously in brief communications, and one is the subject of this paper. Type 1 polyaxonal (PA1) amacrine cells have larger cell bodies than most amacrine cells in Golgi preparations, averaging about 13 microns in diameter. These are typically positioned interstitially in the middle of the inner plexiform layer (IPL), although some are also found in the amacrine and ganglion cell layers. Axons and dendrites are broadly stratified in the middle of the IPL, in the vicinity of the a/b sublaminar border. Sparsely branching dendrites have a conventional appearance, branching at a narrow angle, and giving rise to smaller daughter branches, which taper gradually toward their termination. An unusual feature of the dendrites is the zig-zag course of some terminal branches. Clusters of small, pedunculated spines are common on proximal dendrites, and spines are virtually absent on axons. Axons emerge from proximal dendrites within 50 microns of the soma, and more rarely from the soma, in a tapering initial segment, commonly interrupted by one or two large swellings. Subsequent branching is at a wide angle, and the fine caliber is maintained in the transition from parent to daughter branches. The uniform thickness of the axonal branches is interrupted at intervals by boutons en passant. Although the extent of the dendritic tree is large, exceeding 500 microns in radial extent from the cell body, for cells a few millimeters distant from the visual streak, the axonal tree is much larger, and its radial extent is measured in millimeters. PA1 amacrine cells are believed to be polarized in their functional organization, with a primarily recipient dendritic tree and a primarily transmissive axonal tree. PA1 amacrine cells co-stratify with nab cone bipolar cells and with certain small tufted amacrine and ganglion cells at the a/b sublaminar border. The co-stratification of both axons and dendrites at the a/b sublaminar border of the IPL suggests that PA1 amacrine cells are important modulators of neural activity in the middle of the IPL, affecting both ON and OFF responses, and perhaps ON-OFF cells selectively.

Polyaxonal amacrine cells of rabbit retina: size and distribution of PA1 cells.

Type 1 polyaxonal (PA1) amacrine cells have been identified previously in rabbit retina, and their morphological characteristics have been described in detail in the preceding paper. Like other polyaxonal amacrine cells they bear distinct dendritic and axonal branching systems, the latter of which originates in two to six thin, branching axons which emerge from or near to the cell body. Unlike other types of polyaxonal amacrine cells, however, their branching is stratified at the a/b sublaminar border and their cell bodies are most often displaced interstitially in the inner plexiform layer (IPL). This report emphasizes quantitative features of the population of PA1 cells, documented in Golgi-impregnated and Nissl-stained retinas, and provides further evidence in Nissl preparations for the amacrine-cell nature of polyaxonal amacrine cells. The cell bodies of Golgi-impregnated PA1 amacrine cells are relatively large: 12-15 microns in equivalent diameter over the range extending from the visual streak 6 mm into ventral retina. Over the same range, dendritic trees are 400-800 microns in equivalent diameter, but they are much smaller than the axonal arborizations, which extend up to and perhaps beyond 2 mm from the cell body. Interstitial cell bodies appropriate to PA1 cells have been identified in Nissl-stained, whole-mounted rabbit retinas. In the plane of the retina, these are comparable in area to smaller medium-size ganglion cells, but their very pale Nissl staining, high nuclear/cytoplasmic ratio, and absence of nucleolar staining are all characteristics of amacrine cells. Interstitial displacement of presumed PA1 cells is rare in the visual streak, and the frequency of interstitial cells reaches a peak between 1 and 2 mm ventral to the streak. Counts in Nissl-stained retinas and estimates from nearest neighbor analyses in these and in Golgi-impregnated retinas indicate a density of PA1 cells in the range of 15-16 cells/mm2 at about 2 mm ventral to the streak, when an estimated 25% shrinkage of the material is taken into account. Dendritic field overlap, based upon this estimate, is calculated to be about fourfold, while a lower bound to estimates of the overlap of axonal arborizations is nearly an order of magnitude higher. Many similarities are noted in a qualitative and quantitative comparison of PA1 amacrine cells in rabbit and monkey retinas. In assessing the contribution of the structural organization of PA1 amacrine cells to their possible functional role(s), it is notable that their appearance conforms not to amacrine cells as commonly viewed, but to a more conventional model of neuronal dynamic polarization.(ABSTRACT TRUNCATED AT 400 WORDS)

Polyaxonal amacrine cells of rabbit retina: PA2, PA3, and PA4 cells. Light and electron microscopic studies with a functional interpretation.

Polyaxonal (PA) amacrine cells are a new class of amacrine cell bearing one to six branching, axon-like processes that emerge from the cell body or dendritic trees within 50 microns of the cell body. These slender processes of uniform caliber branch at right angles and in many respects closely resemble the axons of Golgi type II cells found elsewhere in the brain. Of the four types of polyaxonal amacrine cell that we have recognized in rabbit retina, two have been described previously in brief communications. One of these, the PA1 amacrine cell with its interstitially displaced cell body, located in the inner plexiform layer (IPL), has been analyzed extensively in two preceding reports. This paper concerns PA2, PA3, and PA4 amacrine cells. Type 2 polyaxonal (PA2) amacrine cells, identified in Golgi preparations of whole-mounted rabbit retinas, have smaller cell bodies (9-14 microns) than the other three types and these are always displaced to the ganglion cell layer (GCL) or the inner border of the inner plexiform layer (IPL). The dendritic fields of PA2 cells are also smaller than those of other PA amacrine cells, and most of their sparse dendritic branching is narrowly stratified at the border of strata (S) 4 and 5. Some members of this more heterogeneous amacrine cell "type" are bistratified, however, and more highly branched with terminal branches rising to end in S1. PA2 amacrine cells bear a scattering of small dendritic spines and may also exhibit complex dendritic appendages arising at the ends of terminal branches in proximal regions of the dendritic tree. PA2 cells emit one to three axons from the proximal dendritic tree, and about half of the cells bear a single axon. Type 3 polyaxonal (PA3) amacrine cells resemble PA1 cells in the large size of their cells bodies (11-16 microns) and dendritic fields, but differ from the latter in placement of cell bodies, which is in the GCL, and dendritic and axonal stratification, which is multistratified, ranging from S4 to S1, with a concentration in S3 or S4 and a variable contribution to S1. PA3 cells differ from PA1 cells in several other respects, including dendritic branching which occurs at higher frequency and is biased toward temporal retina, and in characteristic bristling dendritic spines, clustered in the intermediate regions of the dendritic tree, that are longer, more variable in appearance and more tightly clustered than the small, uniform spines of PA1 cells that are clustered on proximal dendrites.(ABSTRACT TRUNCATED AT 400 WORDS)

Synaptic organization of starburst amacrine cells in rabbit retina: analysis of serial thin sections by electron microscopy and graphic reconstruction.

The synaptic organization of starburst amacrine cells was studied by electron microscopy of individual or overlapping pairs of Golgi-impregnated cells. Both type a and type b cells were analyzed, the former with normally placed somata and dendritic branching in sublamina a, and the latter with somata displaced to the ganglion cell layer and branching in sublamina b. Starburst amacrine cells were thin-sectioned horizontally, tangential to the retinal surface, and electron micrographs of each section in a series were taken en montage. Cell bodies and dendritic trees were reconstructed graphically from sets of photographic montages representing the serial sections. Synaptic inputs from cone bipolar cells and amacrine cells are distributed sparsely and irregularly all along the dendritic tree. Sites of termination include the synaptic boutons of starburst amacrine cells, which lie at the perimeter of the dendritic tree in the "distal dendritic zone." In central retina, bipolar cell input is associated with very small dendritic spines near the cell body in the "proximal dendritic zone." The proximal dendrites of type a and type b cells generally lie in planes or "strata" of the inner plexiform layer (IPL), near the margins of the IPL. The boutons and varicosities of starburst amacrine cells, distributed int he distal dendritic zone, lie in the "starburst substrata," which occupy a narrow middle region in each of the two sublaminae, a and b, in rabbit retina. As a consequence of differences in stratification, proximal and distal dendritic zones are potentially subject to different types of input. Type b starburst amacrines do not receive inputs from rod bipolar terminals, which lie mainly in the inner marginal zone of the IPL (stratum 5), but type a cells receive some input from the lobular presynaptic appendages of rod amacrine cells in sublamina a, at the border of strata 1 and 2. There is good correspondence between boutons or varicosities and synaptic outputs of starburst amacrine cells, but not all boutons gave ultrastructural evidence of presynaptic junctions. The boutons and varicosities may be both pre- and postsynaptic. They are postsynaptic to cone bipolar cell and amacrine cell terminals, and presynaptic primarily to ganglion cell dendrites. In two pairs of type b starburst amacrine cells with overlapping dendritic fields, close apposition of synaptic boutons was observed, raising the possibility of synaptic contact between them. The density of the Golgi-impregnation and other technical factors prevented definite resolution of this question. No unimpregnated profiles, obviously amacrine in origin, were found postsynaptic to the impregnated starburst boutons.(ABSTRACT TRUNCATED AT 400 WORDS)

A new type of wide-field horizontal cell, presumably linked to blue cones, in rabbit retina.

Horizontal cells of vertebrate retina play an important role in the formation of visual receptive field surrounds of bipolar cells, and hence in the centre-surround receptive field organization of retinal ganglion cells. In some retinas, horizontal cells also play a major functional role in the first stage of colour-coding of visual stimuli. We have identified a new type of horizontal cell, called 'type C', in rabbit retina, which unlike type A and type B horizontal cells, contacts only a small fraction of cone photoreceptors, possibly blue cones. The multiple, sparsely branched axons of type C cells are well-positioned to contact bipolar dendrites in a feedforward manner. In summary, we propose (1) that the presence in rabbit retina of 3 types of cone horizontal cell, A, B, and C, may represent a more common pattern in mammalian retinae, shared with many non-mammalian retinas which contain colour-coded neurons, (2) that type C cells connect principally to blue cones, and (3) that type C cells are more common in retinas, such as those of squirrels, and to a lesser extent rabbits, in which blue cones play a major role in the colour coding of visual signals.

A distinct type of displaced ganglion cell in a mammalian retina.

The morphology, dendritic stratification and laminal position of the soma of retinal ganglion cells were analyzed in Golgi preparations and in other rabbit retinas containing cells backfilled from the superior colliculus. Only one type, among 40 Golgi-impregnated types identified, always had its cell body displaced to the amacrine cell sublayer of the inner nuclear layer. The displaced ganglion cell of rabbit retina has a small cell body, very wide dendritic field with sometimes unbranched dendrites extending up to a millimeter from the cell body. The dendritic tree is narrowly stratified just under the amacrine cell bodies in stratum 1, and therefore does not co-stratify with starburst (cholinergic) amacrine cells, but rather with dopaminergic amacrine cells. Its correlate among ganglion cells backfilled from tectum is apparently a very sparse population of small-bodied cells mixed with a variable population of misplaced ganglion cells of varying size and type. The authentic displaced ganglion cell of rabbit retina, unlike the large displaced ganglion cell of birds, is apparently not a directionally selective ganglion cell, and its functional role in vision is presently unknown.

Starburst amacrine cells in cat retina are associated with bistratified, presumed directionally selective, ganglion cells.

Starburst amacrine cells of cat retina are similar in form, though more delicate and less profusely branched, when compared to the starburst/cholinergic amacrine cells of rabbit retina, as identified in Golgi preparations. In both species, type a cells branch in the middle of sublamina a of the inner plexiform layer (IPL), but type b (displaced) starburst amacrine cells of cat branch near the a/b sublaminar border (stratum 3) of the IPL, not in the middle of sublamina b (stratum 4), as do those of rabbit. Nevertheless, in each species, this starburst substratum in sublamina b coincides with the sublamina b-level branching of a bistratified ganglion cell, which in rabbit retina shows directionally selective responses. It is proposed that starburst amacrine cells of cat retina are cholinergic and, as in rabbit retina, make selective connections with on-off directionally selective ganglion cells.

Immunocytochemical staining of cholinergic amacrine cells in rabbit retina.

Cholinergic neurons of rabbit retina were labelled with an antibody against choline acetyltransferase, the synthesizing enzyme for acetylcholine. Two populations of cells are immunoreactive. Type a cell bodies lie in the inner nuclear layer (INL), their dendrites branching narrowly in sublamina a of the inner plexiform layer (IPL), while type b cell bodies lie in the ganglion cell layer (GCL) with dendrites branching in sublamina b of the IPL. The irregular networks of clustered immunoreactive dendrites are similar, but not identical, in the two sublaminae. Type b cells are more numerous than type a cells in central retina. No axons were stained. It appears that the immunoreactive neurons are normally placed and displaced starburst/cholinergic amacrine cells.

Starburst amacrine cells: morphological constancy and systematic variation in the anisotropic field of rabbit retinal neurons.

Starburst amacrine cells of rabbit retina have been characterized previously in terms of their highly distinctive and regular dendritic geometry. They have been identified as probable cholinergic neurons of the retina and have been shown to direct output solely to ganglion cells. The objectives of this paper are to chart the variation of starburst amacrine cells across the retina, to register the morphological features which are held constant for individual cells, and to examine factors which may remain invariant for the population with change in retinal position. Starburst amacrine cells occur as two completely segregated mirror-symmetrical populations, type a and type b cells, separately serving OFF and ON pathways, respectively. They are treated here as two distinct subpopulations with very similar features. A characteristic morphological feature of both types, related to branching pattern and best seen in flat view, is the location of boutons in the distal annular zone. This is the effective zone of synaptic output, which is constant at 50 to 60% of dendritic field area, regardless of the cell's retinal location. Both type a and type b cells exhibit systematic increase in cell body size and dendritic field diameter, and systematic decrease in frequency of branching and of synaptic boutons with perpendicular distance from the visual streak. These rates of increase or decrease fall off considerably at distances greater than about 1.5 mm dorsal and ventral to the visual streak, but at this distance, the dendritic field diameters of cells in dorsal retina are about 65% larger than the diameters of cells in ventral retina. When type a and type b cells are closely compared, they are seen to differ in several respects. Branching patterns of type a and type b cells differ slightly, the latter being more highly branched, and the normalized branching frequency histograms, characteristic for each type, remain constant with changing retinal position. At the same retinal location type a cells always have larger dendritic field diameters than type b cells. This difference is significant in ventral retina, out to a distance of at least 4.5 mm from the streak. The maximum percentage difference in size occurs not at mid-visual streak, but about 1.5 mm ventral to the streak. The population statistics of dendritic field overlap and areal dendritic coverage have been calculated using published data on cell densities. It is concluded that overlap is extraordinarily high (k greater than 25), more than 10 times that calculated for retinal ganglion cells.(ABSTRACT TRUNCATED AT 400 WORDS)

'Starburst' amacrine cells and cholinergic neurons: mirror-symmetric on and off amacrine cells of rabbit retina.

Golgi-impregnated 'starburst' amacrine cells share significant morphological features with cholinergic neurons in rabbit retina. They are mirror-symmetrical about the a/b (OFF/ON) sublaminar border of the inner plexiform layer. Type a starburst amacrines have cell bodies in the amacrine cell layer and dendrites in sublamina a, while type b cells have their cell bodies in the ganglion cell layer and dendrites in sublamina b of the inner plexiform layer (IPL). The two levels of narrow dendritic stratification are precisely those demonstrated by Masland and Mills for cholinergic amacrine cells. The morphological evidence indicates that the duality of ON and OFF pathways is served separately by type b (displaced) and type a starburst amacrine cells, respectively.

On and off pathways through amacrine cells in mammalian retina: the synaptic connections of "starburst" amacrine cells.

The neural architecture of on and off pathways in mammalian retina is described, including the development of ideas leading to an understanding of the bisublaminar organization of the inner plexiform layer of the retina which supports these two pathways. The complexities of bipolar cell contributions are contrasted with the relative simplicity of ganglion cell organization with regard to bisublaminar architecture, and a key role is described for internuncial amacrine cells as specific targets for bipolar cells. Two very different kinds of amacrine cell are considered and compared, both of which mediate bipolar input to ganglion cells. These are the rod (type II) amacrine cell, and the more recently discovered "starburst" amacrine cell, which is apparently cholinergic in function. As different as the wide-field starburst amacrine cells are from the narrow-field rod amacrine cells, they share important features. Both are interposed between bipolar and ganglion cells, and both have segregated regions of presynaptic boutons. They differ, however, in that rod amacrines may perform more specific functions related to receptive field center organization, while the functional role of starburst amacrines may be unrelated to receptive field properties of ganglion cells. The mirror-symmetry of type a and type b (off and on) starburst amacrine cells is described together with their synaptic circuitry. In contrast to the rod amacrine cell the output of starburst amacrines is exclusively to ganglion cells. Others have proposed a dual function for acetylcholine (ACh) in the retina. A unifying hypothesis is briefly sketched here which relates the pharmacology of ACh and the dendritic stratification of starburst amacrine cells to the form and function of ganglion cells. It is proposed that the amount of generalized synaptic excitation received from ACh/starburst amacrine cells by a particular type of ganglion cell is largely a function of co-stratification of the ganglion cell's dendrites with the distal boutons of starburst amacrine cells.

GABAergic amacrine cells in rat retina: immunocytochemical identification and synaptic connectivity.

GABAergic neurons have been identified in light and electron microscopic preparations of rat retina by an immunocytochemical localization of the GABA-synthesizing enzyme, glutamic acid decarboxylase (GAD). GAD-positive neuronal somata are found only in the inner and middle parts of the inner nuclear layer, and GAD-positive neuronal terminals are observed exclusively within the inner plexiform layer (IPL) and the outermost part of the ganglion cell layer. Dense aggregations of GAD-positive terminals alternate with less dense zones to form a lamination of the IPL. GAD-positive terminals contain pleomorphic synaptic vesicles and are the presynaptic elements of conventional synapses onto bipolar and amacrine cell processes, as well as onto the somata and dendrites of ganglion cells. In addition, GAD-positive terminals are postsynaptic to unstained bipolar terminals and are components of synaptic dyads where they occasionally appear to form reciprocal synapses with the bipolar terminals, and serial with unstained amacrine processes. Probable synaptic contacts between adjacent GAD-positive terminals also have been observed. Most of the synaptic input to GAD-positive terminals comes from bipolar cells, while the small remaining input mainly comes from other GAD-positive terminals. The synaptic output to GAD-positive terminals is greatest to bipolar cells, followed in decreasing order by GAD-negative amacrine cells, ganglion cells, and other GAD-positive cells. The total synaptic output of GAD-positive cells appears to be more than twice as great as the total input of these cells. The location of GAD-positive somata, the distribution of GAD-positive terminals, and the synaptic relationships formed by these terminals all indicate that amacrine cells are the only GABAergic neurons in rat retina. Our observations also indicate that not all amacrines are GABAergic and suggest that GABAergic neurons may be limited to a narrow field subclass of amacrine cell. The findings concerning the synaptic connections of GABAergic amacrines suggest that such cells are the first link in several divergent pathways from bipolar to ganglion cells and that they probably serve more than one function since they feed synaptic activity forward directly upon ganglion cells as well as back upon bipolar cells.

Golgi-impregnated amacrine cells and GABAergic retinal neurons: a comparison of dendritic, immunocytochemical and histochemical stratification in the inner plexiform layer of rat retina.

This paper concerns the banding pattern produced in the inner plexiform layer of rat retina by glutamic acid decarboxylase (GAD) immunocytochemistry. It presents a comparison of this pattern with the dendritic stratification of neurons that are reasonable candidates for GABAergic amacrine cells in Golgi preparations, and also with the banding patterns produced by other histochemical techniques. First, the spacing of five dense GAD-positive bands and four intervening less dense bands in central retina is quantitatively described. Second, examples of a particular, morphologically homogenous group of Golgi-impregnated amacrine cells are examined in the details of their structure, especially with regard to their dendritic stratification. Computer reconstructions of the dendritic trees of some of these narrow-field, multistratified amacrines are compared with the GAD-positive banding pattern. This group of amacrines is judged to represent many of the GABAergic neurons in rat retina, accounting for the form and distribution of GAD-positive synaptic terminals by their dendritic morphology and stratification. Third, a general schema for the laminar subdivision (stratification) of the inner plexiform layer in rat retina is derived from a comparison of the results of several histochemical procedures. Finally, similarities and differences in the distribution of GAD-positive amacrine cell dendrites are noted among mammals and the functional implications of their broad distribution are discussed. A conspicuous difference is cited between mammals and certain nonmammalian vertebrates in which GAD-positive dendrites are restricted to sublamina beta (ON-center cells) of the inner plexiform layer.