AII Amacrine Cells Research Articles

Pharmacological modulation of the rod pathway in the cat retina.

1. In the intact cat eye, the responses of ganglion cells to light stimulation were recorded extracellularly and the actions of iontophoretically applied 2-amino-4-phosphonobutyrate (APB), a potent agonist at ON-bipolars, and of strychnine, a glycine antagonist, were investigated. 2. Under light-adapted conditions, the activity of ON-center ganglion cells is decreased by APB but is increased by strychnine. APB and strychnine act independently of one another. 3. The activity of light-adapted OFF-center ganglion cells is increased by APB and by strychnine. The light response remains clearly modulated. Strychnine blocks the action of simultaneously applied APB. The results are in agreement with the action of a push-pull mechanism, according to which ON-cone-bipolars provide a glycinergic input into OFF-center ganglion cells. 4. Under dark-adapted conditions, APB blocks the light responses of both ON-center and OFF-center ganglion cells. The discharge rate of ON-center ganglion cells is completely suppressed; OFF-center ganglion cells show a high maintained discharge. 5. Strychnine blocks the scotopic light response of OFF-center ganglion cells and blocks the action of simultaneously applied APB. The light response of ON-center ganglion cells is hardly affected by strychnine. 6. The effects of strychnine on OFF-center ganglion cells are in agreement with the hypothesis that the glycinergic AII amacrine cells modulate the activity of the scotopic OFF-channel. 7. Intravitreally applied APB abolished the scotopic b-wave of the electroretinogram at concentrations of 100 microM. 8. Our data suggest that as in rabbit (10) the rod bipolars in cat retina are depolarizing (ON) bipolar cells.

A17: a broad-field amacrine cell in the rod system of the cat retina.

A17 amacrine cells of the cat retina have been penetrated with horseradish peroxidase (HRP)-filled microelectrodes and their light responses recorded. These cells depolarize in sustained fashion to steps of light. Viewed in retinal wholemounts, HRP-injected cells have a spokelike radiating splay of very fine dendrites (0.1 micron diam) passing diffusely through all strata of the inner plexiform layer (IPL) to run primarily in strata 4 and 5. There are as many as 1,000 large, regularly spaced beads borne on the 500- to 1,200-micron diameter dendritic field. Cell body sizes range from 9 to 13 micron. In the electron microscope, the dendritic beads in sublamina b of the IPL are seen to synapse reciprocally with rod bipolar axon terminals. Dendritic beads in sublamina a rarely make synapses, but between the beads in this layer, input from at least three distinctive amacrine profiles occurs. Though diffuse at the light microscopic level, A17 thus appears to be structurally bistratified, with amacrine input in sublamina a and bipolar input in sublamina b. It is likely that A17 can be identified with AI. A17 signals are driven almost exclusively by rods. The spectral sensitivity peaks at 507 nm, identical with that of pigment epithelial cells. Light adaptation abolishes all but a small hyperpolarizing component of the signal. The overall intensity-response range is similar to that of AII amacrine cells. When receptive fields of A17 cells are mapped with slit stimuli, a broad, single-component curve is measured approximately covering the dendritic field. The receptive field is well described by a linear electrical model with a mean space constant of 259 +/- 97 micron (SD). On the other hand, responses to centered slit stimuli of varying width yielded space constants of only 38 +/- 29 micron. A17 amacrines are thus broad-field components of the cat's rod system but with very little capacity for spatial integration. Receptive-field measurements are not supportive of the notion of isolated dendritic regions.

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.

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.

Rod input to amacrine cells in dace retina

Intracellular recordings in dace retina demonstrate a type of amacrine cell whose responses to flashes of light were characteristically a transient epolarization at the onset of light, followed by a slow decay toward the dark level, similar to rod-dominated, AII amacrine cells described in cat retina. Electron microscopy of HRP-filled dace AII-like amacrine cells suggested that they receive significant rod inputs through rod-dominated bipolar cells.

Synaptic patterns and response properties of bipolar and ganglion cells in the cat retina

After intracellular recording, bipolar cells of the cat retina have been stained with HRP and their contacts in the outer and inner plexiform layers examined by electron microscopy. Rod bipolars and cone bipolar cb6 make invaginating, ribbon related contacts with photoreceptors, hyperpolarize in response to light, and have axons terminating in layer b of the IPL. The axon terminal of cb2 ends in layer a of the IPL and its basal contacts with cones mediate hyperpolarizing light-responses. Cone bipolar cb5 is a center-depolarizing type with an axon ending in layer b but its cone contacts are at semi-invaginating basal junctions. Except for the amacrine-contacting rod bipolar cell, all cone bipolar types synapse with both amacrine and ganglion cells in the inner plexiform layer. In addition cb5 contacts AII amacrine cells with large gap junctions, and is physiologically rod dominated.

Dopaminergic amacrine cells in the cat retina

Autoradiographic studies of cat retina showed an accumulation of [ 3H]dopamine in a subpopulation of amacrine cells whose process ramify in the outermost stratum of the inner plexiform layer. Dendrites of these cells are characterized by numerous varicosities measuring up to 2 μm in diameter which are connected by fine intervaricose segments. The dopaminergic amacrine cells are presynaptic to other labeled cells and to unlabeled amacrine populations but not to bipolar or ganglion cells. [ 3H]Dopamine-labeled processes provide extensive synaptic input to the somata and lobular appendages of type AII amacrine cells. This relationship suggests that dopaminergic amacrine cells may play an important role in the regulation of rod pathways in the cat retina.

AII amacrine cells quicken time course of rod signals in the cat retina.

1. In a perfused eyecup preparation, AII amacrine cells of the cat retina were penetrated with glass microelectrodes and their electrical responses to photic stimuli recorded. 2. Intracellular injections of the stains Procion, lucifer, or horseradish peroxidase revealed dendritic tree diameters of 30-80 micrometers (48 +/- 16 micrometers, mean +/0 SD) and cell body diameters from 7 to 12 micrometers (9 +/- 3 micrometers) for these cells. The dendrites were broadly stratified throughout the inner plexiform layer (IPL) but possessed large, terminal varicosities in the IPL and inner nuclear lyer (INL) proximal to the cell body. 3. The waveform of these cells in response to photic stimulation suggested division into four components: a) an initial rapid depolarization of the cell membrane followed by a slower decay toward the dark level; b) suppression of the dark noise of the cell; c) with dim or moderately intense stimuli, an off-hyperpolarization; d) in some cases a hyperpolarizing surround response. 4. The receptive fields of AII cells have been characterized using spatial stimuli consisting of long narrow slits. Curves have been fitted to spatial data using two space constants, one for the center mechanism and an opposing one for the surround. For the central mechanism, space constants ranged from 20 to 80 micrometers (46 +/- 22 micrometers), while for the surround they ranged from 60 to 130 micrometers (85 +/- 28 micrometers). The mean half-width of the center mechanism, calculated from the mean space constant, was about 0.25 degrees of visual angle (64 micrometers). The receptive-field properties of AII amacrine cells resemble those of center-depolarizing bipolar cells of other species. 5. Spectral studies of AII amacrine cells reveal that they are rod driven at all criterion voltage levels. Furthermore, adaptation of the rods by rod-saturating backgrounds eliminates 95% of the response amplitude of the AII amacrine cells. Under these conditions the tiny response component remaining is driven by the cat's long wavelength (556-nm peak) cones. 6. AII amacrine cells depolarize to rod stimulation more rapidly than other rod-dominated cells, such as rod bipolar cells, which hyperpolarize. For stimuli corresponding to about 10% of rod saturation, the latency to half-maximum amplitude is about 65 ms for AII cells, 40 ms faster than rod-dominated hyperpolarizing units. The leading edge of the response waveform for AII cells is also much more restricted in time. With the above stimulus it requires about 20 ms to increase from 25 to 75% of its peak, a period almost 4 times shorter than required by rod-dominated S-potential responses. With saturating stimuli the AII response requires only 5 ms to increase from 25 ot 75% of its peak. 7. Although prominent in the rod system, AII amacrine cells do not appear to be able to detect single quantum events. Threshold signals require the bleaching of about 200 rhodopsin molecules within a receptive field containing some 1,300 rods...