Localization of aspartate-like immunoreactivity in the retina of the turtle (Pseudemys scripta).

Aspartate has been reported to be a putative excitatory neurotransmitter in the retina, but little detailed information is available concerning its anatomical distribution. We used an antiserum directed against an aspartate-albumin conjugate to analyze the anatomy, dendritic stratification, and regional distribution of cell types with aspartate-like immunoreactivity in the turtle retina. The results showed dramatic differences in immunoreactivity in the peripheral versus the central retina. Strong aspartate-like immunoreactivity was shown in the peripheral retina, with many well-labeled processes in the inner plexiform layer. Many bipolar, horizontal, amacrine, and ganglion cells, some photoreceptors, and some unidentified cells were strongly immunoreactive in the peripheral retina. In contrast, although the central retina showed well-labeled horizontal cells, there was only light labeling in the inner plexiform layer with weakly immunoreactive amacrine and ganglion cells and no labeled bipolar cells. There were several strongly immunoreactive efferent nerve fibers which left the optic nerve head and arborized extensively in the retina. At the electron microscopic level, electron-dense reaction product was associated with synaptic vesicles at bipolar and amacrine cell synapses in the inner plexiform layer. These results suggest that aspartate may be involved in many diverse synaptic interactions in both the outer plexiform layer and the inner plexiform layer of the turtle retina.     

Fluorescence and electron microscopical observations on the amine-accumulating neurons of the cebus monkey retina

The organization of the Cebus monkey regina was analysed after the intraocular injection of 5,6-dihydroxytryptamine. This amine was taken up not only by the previously known dopaminergic neurons, but also by a set of indoleamine-accumulating neurons, whose processes are confined to the inner plexiform layer. The synaptic contacts of the dopaminergic neurons were analysed in the electron microscope after the processes of the indoleamine-accumulating neurons were destroyed by the intravitreal injection of the neurotoxic indoleamine, 5,7-dihydroxytryptamine. The subsequent injection of 5,6-dihydroxytryptamine induces certain changes in the dopaminergic neurons which accumulate the substance: electron-dense cores appear in the synaptic vesicles, and increased electron-density of mitochodrial and cellular membranes is often observed. The dopaminergic neurons were found to be presynaptic to amacrine cell perikarya and processes in the inner plexiform layer. In the outer plexiform layer they were presynaptic to both bipolar and horizontal cells, but they did not contact photoreceptors. The dopaminergic neurons received synapses only in the inner plexiform layer, from amacrine cell processes. It is inferred that in Cebus most dopaminergic neurons belong to a special class of retinal neuron, the interplexiform cells, which appear to transmit information centrifugally within the retina, from the inner to the outer plexiform layers. There are considerable similarities between the synaptology of the dopaminergic interplexiform neurons in the Cebus monkey and the goldfish retina, and the function of interplexiform neurons may therefore be similar in these two species.     

Tyrosine hydroxylase-immunoreactive elements in the distal retina of Bufo marinus: a light and electron microscopic study.

Tyrosine hydroxylase-immunoreactive elements in the distal retina of Bufo marinus were investigated using light and electron microscopic immunocytochemistry. At the light microscopic level, immunoreactive somas were seen in the proximal part of the inner nuclear layer, and immunoreactive processes projected both to the inner and outer plexiform layers. In some instances stained axon-like processes traveled from the inner plexiform layer, across the inner nuclear layer to the distal retina. Immunolabeled elements formed basket-like structures around the photoreceptor inner segments. At the ultrastructural level immunostained fibers were observed in close contact with the necks, lateral walls, bases and the outer surfaces of rod outer segments. Synaptic specializations were neither observed at rod contacts nor at other possible contact sites such as bipolar dendrites and horizontal cell somata and processes in the outer plexiform layer. In contrast, synaptic specializations between immunolabeled profiles and amacrine, bipolar and ganglion cells were regularly present in the inner plexiform layer. These findings suggest that a population of dopaminergic interplexiform cells is present in the Bufo retina and sends axon-like processes towards the distal retina. It is assumed that dopamine is probably released non-synaptically from the immunolabeled terminals in the distal retina influencing rods directly, by which the quality of photopic vision is enhanced in the anuran retina.     

Light microscopy of GTP-binding protein (Go) immunoreactivity within the retina of different vertebrates

To examine species differences in the distribution pattern of guanosine triphosphate (GTP)-binding protein (Go) within the vertebrate retina, paraffin-embedded retinae from a number of vertebrate species, including the goldfish, frog, turtle, chicken, monkey, and human, were immunohistochemically stained with affinity-purified antibody against the alpha-subunit of Go. Go-immunoreactive products were found to be located in the neuropil, but not in the cell bodies of neurons, in the retina of all these species. However, some species differences were observed. In the frog, monkey and human, the inner plexiform layer (IPL) was homogeneously stained with this antibody, but in the goldfish, turtle and chicken, the IPL was heterogeneously stained. In the frog, chicken, turtle and human, the outer plexiform layer (OPL) was densely stained with this antibody, but in the goldfish and monkey, the OPL was rather faintly immunoreactive to the antibody. In the goldfish, monkey and human, the outer nuclear layer (ONL) was not immunoreactive to the Go-antibody, whereas in the frog, turtle and chicken, the ONL was immunoreactive to it. The implications of these species differences in Go localization in the vertebrate retina are discussed.     

Postnatal development of dopamine D1 receptor immunoreactivity in the rat retina.

Dopamine, an important neuromodulator in the retina, controls the balance of rod cone photoreceptor activity and influences the activity of several interneurons. The postnatal development of dopaminergic neurons, visualized immunocytochemically, was compared to the development of dopamine D1 receptor immunoreactivity. Expression of D1 receptors was monitored throughout the postnatal development of the rat retina using a subtype-specific monoclonal antibody. D1 receptors are expressed in the inner plexiform layer beginning at birth. Labeling of the inner plexiform layer changed from a diffuse pattern, staining the entire layer, to the typical adult punctate staining, that was organized in layered bands and occurred in the second postnatal week. The staining did not co-localize with dopaminergic cells; instead, it colocalized with cells in the inner nuclear layer or the ganglion cell layer. Within these cells, D1 receptors were most heavily expressed in processes stratifying in the inner plexiform layer. Staining in the outer plexiform layer and in horizontal cells was found beginning in the second postnatal week. Clustering of the D1 receptor within plexiform layers, a process typical for the well-described function of dopamine modulation in the adult, occurred late in postnatal development. A possible function of D1 receptors in neuronal development is discussed.     

Cholinergic and GABAergic neurons occur in both the distal and proximal turtle retina.

Turtle retinas were processed immunocytochemically and histochemically to detect the presence of choline acetyltransferase (ChAT), acetylcholinesterase (AChE), and glutamate decarboxylase (GAD). We observed cholinergic and gamma-aminobutyric acid (GABA)ergic neurons in the proximal retina, as expected, and in the distal retina as well. ChAT immunoreactivity in the distal retina was observed within the axons and pedicles of numerous cone photoreceptors, suggesting that a population of turtle cone photoreceptors uses ACh as a neurotransmitter. Type L2 horizontal cells were immunoreactive for GAD, and their dendrites invaginated into cone pedicles. AChE histochemistry revealed processes within the outer plexiform layer which formed a loosely organized lattice. In the proximal retina, labeling for ChAT and GAD was similar to that reported by previous investigators. Processes from ChAT-labeled amacrine cells in the inner nuclear layer formed a stratum within the distal inner plexiform layer (IPL) (at 16-21% relative IPL depth), and processes from ChAT-labeled amacrines in the ganglion cell layer formed a proximal ChAT stratum (at 55-58% relative IPL depth). In addition, six AChE-labeled bands and five GAD-labeled bands were observed within the IPL of stained retinas. Therefore, we determined that the two broadest AChE-labeled bands and the two broadest GAD-labeled bands overlapped the two labeled ChAT strata. The evidence for cholinergic and GABAergic processes in both the inner plexiform layer and the outer plexiform layer, combined with electrophysiological evidence from other investigators, raises the possibility that distal retinal neurons may be involved in the encoding of directional information.     

Identification of the interplexiform cell in the dace retina by dye-injection method

The responses of interplexiform cells in the dace retina were recorded intracellularly and identified morphologically. The response pattern closely resembles the response of bipolar cells and amacrine cells (on- and off-types). Morphologically, the perikaryon of most of these cells lies in the amacrine cell layer, and is usually large in size. Distal and proximal processes arise from the soma directly. The distal process ramifies and extends widely in the outer plexiform layer, but does not make contact with receptor terminals. The proximal process branches and extends widely in the inner plexiform layer. The majority of these cells resembles the dopaminergic interplexiform cells, but a few resemble the glycinergic interplrxiform cells in the goldfish retina.     

Neuronal markers in rat retinal grafts

Rat E15 retina was grafted to the retina of adult rat hosts. After varying survival times (1 week-6 months), grafts were stained by immunohistochemistry for neurofilament 160 kDa (NF), HPC-1 (an amacrine cell marker), choline acetyltransferase (ChAT), tyrosine hydroxylase (TH), glutamic acid decarboxylase (GAD) and somatostatin-28 (SS-28). The first differentiating graft amacrine cells (cholinergic and dopaminergic) could be seen 1 week after transplantation (corresponding to postnatal day 1 = P1). The inner plexiform layer of the graft started to differentiate at 2 weeks (corresponding to P8) seen by HPC-1 and GAD staining. ChAT, TH and SS-28 immunostaining revealed an abnormal lamination pattern in the graft inner plexiform layer. Also by 2 weeks, the outer plexiform layers of the graft contained NF-immunoreactive horizontal cells. No NF-stained retinal ganglion cells could be observed in the graft. Five and 7 weeks after grafting, the transplants had obtained the same staining intensity with different markers as the host retina.     

Neuropeptide Y-immunoreactive amacrine cells in the retina of the turtle Pseudemys scripta elegans

Antiserum directed against neuropeptide Y selectively labeled certain amacrine cells in the turtle retina. The cell types, sizes, dendritic stratification, regional distribution, and degrees of immunolabeling were examined. The results indicated that three morphologically distinct cell types were labeled: types A, B, and C. Computer rotation of digitized data from camera lucida drawings was used to study dendritic stratification. The type A somata were large (11.5 micron in diameter), well-stained, and located in the third tier of the inner nuclear layer. Type A somata gave rise to well-stained processes which arborized within the inner plexiform layer in strata 1 and 3 and at the border between strata 4 and 5. Processes in stratum 1 were sparse and delicate with small boutons. Processes in stratum 3 were numerous and often coarse, with many large and small boutons. At the border between strata 4 and 5 the processes were frequently numerous but slender, with many small boutons. Occasional immunolabeled processes were found in the ganglion cell layer. The somata of the type B cells were smaller (9.0 micron in diameter) and gave rise to single labeled processes which descended into the inner plexiform layer and divided quickly into two secondary processes. These secondary processes gave rise to lightly labeled dendritic fields which arborized primarily in strata 2 and 4. The type C cells were usually observed at the periphery of the retina and had large somata (12.0 micron in diameter) with simple, but very elongated, dendritic arborizations in strata 1, 4, and 5. Observations also showed that type A and B cells were often found in close proximity to each other and suggested that dendrites of these cells made contact with each other. The labeled neurons were distributed relatively evenly throughout the retina except for the visual streak where they were sparse. This study indicates that neuropeptide Y-like immunoreactivity is found in more than one anatomically distinct class of amacrine cells in the turtle retina.     

Dopaminergic periglomerular cells in the turtle olfactory bulb

With the indirect immunofluorescence technique using antisera to three catecholamine synthesizing enzymes, labeled periglomerular cells as well as their intraglomerular processes were observed in the turtle olfactory bulb. These cells could also be recognized in the EPL and the glomerular layer. Unlabeled periglomerular cells were also seen. Thick labeled processes (presumably dendrites) entered the glomerular neuropil, and there formed a dense network, with numerous terminal varicosities. These results support the existence of a unique, homologous dopaminergic subdivision of the periglomerular interneurons throughout classes of vertebrates. In addition, a second type of weakly tyrosine hydroxylase immunoreactive neurons was observed in the outer part of the granule layer. Dopamine beta-hydroxylase positive fibers were seen in the granule, mitral and external plexiform layers.     

Dopaminergic neurons in the human retina

The utilization of dopamine in the adult human retina was examined by using high-affinity uptake, localization, synthesis, and release as neurotransmitter-specific physiological probes. Autoradiographic and histochemical studies have shown that dopamine-accumulating and dopamine-containing cells of the human retina belong to a population of neurons whose somata are located in the proximal regional of the inner nuclear layer. Some of these are amacrine cells which are pre- and postsynaptic to other amacrine cells exclusively in the inner plexiform layer. However, evidence is presented which indicates the existence of interplexiform dopaminergic neurons which send processes to both plexiform layers of the retina. These neurons contain a high concentration of dopamine, take up 3H-dopamine by a hig-affinity mechanism, and release endogenous or accumulated dopamine by a Ca2+-dependent mechanism upon depolarization with high extracellular K+. An endogeneous level of about 20 pmoles dopamine per mg protein was measured in freshly isolated retina using high-pressure liquid chromatography with electrochemical detection. These results demonstrate that mechanisms for dopaminergic neurotransmission are present in the human retina.     

Morphology and distribution of catecholaminergic amacrine cells in the cone-dominated tree shrew retina.

The tree shrew (Tupaia belangeri) has a cone-dominated retina with a rod proportion of only 5%. This is in contrast to the usual mammalian pattern of rod-dominated retinae. Rod bipolar cells are present at relatively low densities in the tree shrew retina, suggesting that a reduced, but normal, rod pathway might be preserved. The present study investigated another common constituent of the rod pathway, the dopaminergic amacrine cells, and analysed their morphology and distribution by light and electron microscopy. Catecholaminergic (presumed dopaminergic) amacrine cells were labelled with an antibody against tyrosine hydroxylase (TH). Intense TH-immunoreactivity was found in perikarya and dendrites of a uniform amacrine cell population. TH-immunoreactive amacrine cell density varies across the retina from 10 cells/mm2 in the periphery to 40 cells/mm2 in more central regions (mean cell density about 25 cells/mm2). The relatively large cell bodies are located exclusively in the innermost part of the inner nuclear layer. The dendrites form a dense plexus at the border between the inner plexiform layer and the inner nuclear layer. The finer dendritic processes contain many varicosities and form characteristic dendritic "rings" like those seen in other mammals. TH-immunoreactive processes also run between cell bodies in the vitread inner nuclear layer; a few extend into the sclerad inner nuclear layer and occasionally reach the outer plexiform layer (possible interplexiform cells). A few TH-immunoreactive processes are seen in the middle of the inner plexiform layer. Electron microscopy of TH-immunoreactive processes revealed conventional synapses onto somata and processes of unlabelled amacrine cells.     

Interplexiform cells of the goldfish retina

Dopaminergic and glycinergic interplexiform cells (IPCs) in the goldfish retina were impregnated by using two new Golgi protocols. The two cell types have markedly different morphological characteristics: Dopaminergic IPCs have primary dendrites that descend into and stratify in the inner plexiform layer, where they give rise to processes that project to the outer plexiform layer. Conversely, glycinergic IPCs have primary dendrites that ascend to the outer plexiform layer and from this dendritic arbor, many processes then project into the inner plexiform layer. The apparent coverage of dopaminergic IPCs is almost four times that of glycinergic IPCs. Even so, the coverage of each glycinergic IPC in the outer plexiform layer allows it to provide an accurate copy of the S-space to the inner plexiform layer. Considering the known GABAergic and glycinergic synaptologies in the inner plexiform layer, the glycinergic IPC must form a major element in the retinal circuitry of the goldfish.     

Serotonin and dopamine in the retina of a lizard

The objective of this paper is to report the presence and localization of serotonin and dopamine in the retina of the lizard Uta stansburiana. High performance liquid chromatography and electrochemical detection were used to identify and quantitate the two amines. Both compounds are present as endogenous molecules in this retina and are found in concentrations similar to those reported in other non-mammalian retinas. The same methods were employed to confirm, in the isolated retina, the synthesis of serotonin from precursor, tryptophan. Immunocytochemical methods were used to localize, in the neural retina, serotonin and the rate-limiting enzyme of dopamine synthesis, tyrosine hydroxylase. Serotonin immunoreactivity was observed in bistratified amacrine cells (ca. 7 micron dia.) with processes ramifying in sublayers 1, 4, and 5 of the inner plexiform layer. Immunoreactivity to tyrosine hydroxylase was observed in a different population of bistratified amacrine cells (ca. 11 micron dia.) that had processes ramifying in sublayers 1 and 5 (and perhaps 3) of the inner plexiform layer. The enzymes for further metabolism of dopamine were not found in the retina of this lizard by immunocytochemical methods. The results of this research suggest that only single classes of serotoninergic and dopaminergic neurons are present in the retina of U. stansburiana. This retina might, therefore be an appropriate place in which to investigate the functioning of these amines in visual information processing.     

Distribution and spatial geometry of dopamine interplexiform cells in the rat retina: I. Developing retina

The morphology and distribution of dopaminergic interplexiform cells were analyzed in 9-day-old rat retinas processed as wholemounts for tyrosine hydroxylase immunohistochemistry. The mean number of dopaminergic interplexiform cells was estimated as about half of the total number of dopaminergic neurons in the immature retina, with a higher density in the temporal retina. Four interplexiform cells were individually analyzed and reconstructed with a computer system. Their arborizations could be divided into three different regions based on both their morphological features and their position within the retinal layers: (1) an internal arborization, spreading at the margin between the inner nuclear layer and the inner plexiform layer, composed of long, thick, somatofugal dendrites branching at acute angles, (2) an external arborization in the middle of the inner nuclear layer, formed by short, thin, varicose, recurved, axon-like processes branching at right angles, (3) and one or more scleral process(es), originating either from the cell body or from the internal arborization, running toward the outermost cell row of the INL, some of which reached the outer plexiform layer. Finally, analysis of the arborization network by computer simulations based on the 4 digitalized cells was compared with a nearest-neighbour analysis of cell body distribution. It showed that cell bodies are almost randomly distributed--at least in the inferior retina--but that an adjustment of dendritic growth and orientation probably occurs to ensure a homogeneous coverage of the retina with a constant degree of overlap in the adult. This report represents the first three-dimensional computer reconstruction of chemically identified neurons in the retina.     

Shapes and distributions of the catecholamine-accumulating neurons in the rabbit retina

Rabbit retinas were fixed with mixed aldehydes and examined for the fluorescence of catecholamines. Labeled cell bodies were present in the layer of the amacrine cells. A band of fluorescent processes was present in layer 1 of the inner plexiform layer. Weaker labeling was present in two deeper strata, one near the middle of the inner plexiform layer (presumably layer 3) and one at the juction of layers 4 and 5. Immunohistochemistry showed tyrosine hydroxylase (TH) to be present in the same cells and the same strata of the inner plexiform layer as the endogenous catecholamines. Exposing the retina to exogenous dopamine or norepinephrine resulted in stronger labeling in the middle and deep levels of the inner plexiform layer. At the same time a second population of amacrine cell bodies became visible. Catecholamine fluorescence contained in the amacrine cell bodies was used as a guide to their injection with Lucifer yellow CH. The filled dendritic arbors revealed two main types of cells. The type 1 cells are monostratified at the most distal level of the inner plexiform layer. They have relatively uncomplicated, radially branching dendritic trees. They are the cells densely stained by immunohistochemistry with antibodies against. TH. The type 2 cells are tristratified, with minor branching in layer 1 of the inner plexiform layer and major branching in the two deeper sublayers. The descending dendrites follow a complicated course, and it is not uncommon for intermediate dendrites to cross between strata more than once. The relationship of the cells to their dendritic plexuses was further studied in retinas in which the aldehyde-induced fluorescence of catecholamines was photoconverted to a diaminobenzidine product. The type 1 cells were found to dominate the plexus of dendrites in layer 1 of the inner plexiform layer. The catecholaminergic plexuses in the middle and deep levels of the inner plexiform layer are formedf by dendrites of the type 2 cells. The position of every type 1 cell was mapped in retinas stained with antibodies directed against TH. In one retina we counted 5,613 type 1 cells, distributed evenly across the retina. In another retina, all of the catecholamine-accumulating cells were counted. There were 9,058 with a distribution that peaks in the visual streak. The type 1 cells appear to be the dopaminergic cells previously studied by others and thought to regulate the flow of information from rod bipolar cells to ganglion cells. The low density and wide spread of type 2 cells suggests that they, too, perform a generalized control function, presumably a novel one that dictates their intricate, tristratified shape.     

The dopamine D2 receptor subfamily in rat retina: ultrastructural immunogold and in situ hybridization studies

Dopamine, a major neurotransmitter in the vertebrate retina, is released from interplexiform cells and a restricted subset of amacrine cells. Dopamine effects vary between different retinal cell types, most likely due to differences in cell-specific receptor subtype expression. Identification of cells expressing receptors of the D2-subfamily (D2R, D3R, D4R) on a light microscopical level has rendered equivocal results, and no information is as yet available concerning the subcellular distribution of receptor protein. In the present study, D2R and D2/3R subtype-specific antisera, and D2R-, D3R- and D4R-specific oligonucleotide probes were used for ultrastructural and in situ hybridization analyses of the receptor subtype distribution in the rat retina. Light and electron microscopy showed that in addition to the known localization of intense D2R-immunoreactivity in all dopaminergic cells immunoreactive for tyrosine hydroxylase (TH), homogeneous, less intense D2R-immunoreactivity was also seen throughout the inner plexiform layer (IPL). Ultrastructurally, many additional amacrine cell processes devoid of TH-immunoreactivity at all levels of the inner plexiform layer were immunoreactive. D2R-immunoreactivity was found mainly on intracellular vesicles, and immunoreactivity associated with the plasma membrane was always extrasynaptic. No D2R-immunoreactivity was found in amacrine cell somata postsynaptic to the so-called dopaminergic 'ring endings'. Many D2R-mRNA reactive cells were observed throughout the inner nuclear layer. Morphologically, labelled cells resemble amacrines and bipolars but not horizontal cells. Reactivity with splice variant-specific oligonucleotide probes suggested that the D2LR variant is the predominant if not the only D2R isoform in the rat retina. D2R-mRNA reactivity was not observed in other retinal layers, in particular not in photoreceptor inner segments, which displayed D4R-mRNA reactivity. D3R-mRNA reactivity was not detected. The results indicate that D2-like responses are mediated through the D2R subtype, by an autoreceptor mechanism in dopaminergic cells, and by volume transmission in non-dopaminergic cells of the inner retina. D2-like responses in photoreceptors probably represent D4R activation.     

Distribution and spatial geometry of dopamine interplexiform cells in the retina. II. External arborizations in the adult rat and monkey

The morphology and distribution of dopaminergic interplexiform cells in adult rat and monkey retinas were analyzed to determine any correlation with the function of dopamine in the outer retinal layers. The retinas were processed as whole mounts for tyrosine hydroxylase immunohistochemistry. There was a network formed by the sclerally directed processes of interplexiform cells in the inner nuclear, outer plexiform, and outer nuclear layers running throughout the retina. Their density was higher in the superior retina than in the inferior retina of the rat and was especially high in the superior temporal quadrant. The external network in this quadrant was significantly less dense in the monkey than in the rat, as are the interplexiform cells. The somata of interplexiform and other dopaminergic cells were about the same size in both rats and monkeys. Computer-assisted reconstruction of external arborizations of individual cells showed that external processes lay very close to horizontal and photoreceptor cells and also to blood capillaries. Because they were long, thin, and highly varicose; branched at right angles; and often arose from an axon hillock, the external processes were identified as axons. Therefore, we define the dopaminergic interplexiform cells as multiaxonal neurons, with at least one outwardly directed axon that reaches the outer plexiform layer. The function of the network of external processes from the interplexiform dopaminergic cells is discussed in terms of modulating the release of dopamine to external layers.     

Distribution of immunoreactivity to protein kinase C in the turtle retina

Immunocytochemical staining procedures using the HRP-complexed antibody to protein kinase C (PKC) have been carried out on the turtle retina. Wholemounts and frozen sections of retina have been studied by light microscopy to evaluate PKC immunoreactivity after stimulation of the retina with light and neurotransmitters known to be active in the vertebrate retina. The most dramatically stained sites are cone synaptic pedicles and bipolar cells under all conditions. Ganglion cells stain weakly under certain conditions. Applying the antibody to a 'control' retina under dark adapted conditions results in uniform background staining of both hyperpolarizing and depolarizing bipolar pathways, while stimulating the retina with K+ under dim light conditions results in discretely stained bipolar cells and a prominent band of staining in stratum 4 of the inner plexiform layer. Stronger stimulation of bipolar cells with their terminals contributing to strata 3 and 4 and the continuous dominant band in stratum 4 can be elicited with incubation of the retina in neurotransmitter agonists, GABA and dopamine. Incubation with dopamine, in particular, brings out the putative dopaminergic amacrine cell. The only condition in which a strong band in stratum 2 can be demonstrated is under stimulation with a flashing bar of spot of light. Thus K+ and neurotransmitter stimulation elicit PKC staining in neurons contributing to the ON or depolarizing sublamina of the IPL, while intermittent flashing light stimulus is required to elicit PKC staining in the OFF or hyperpolarizing sublamina of the IPL.     

Retinal dopaminergic neurons (A17) expressing neuromedin K receptor (NK(3)): a double immunocytochemical study in the rat

By using a double immunofluorescence method we examined the distribution of dopaminergic neurons (A17) expressing neuromedin K receptor (NKR, NK(3)) in the rat retina. The distribution of NKR-like immunoreactive (-LI) neurons partially overlapped that of tyrosine hydroxylase (TH)-LI neurons in the inner retina of section and flat-mount preparation. Neurons showing both TH- and NKR-like immunoreactivities were found in the retina (A17): 100% of these TH-LI neurons displayed NKR-like immunoreactivity, and they constituted about 3.5% of total NKR-LI neurons. The majority of double-labeled neurons with TH- and NKR-like immunoreactivities were distributed in the proximal inner nuclear layer and the upper part of inner plexiform layer of the retina, and characterized with appearance of amacrine cells. The present study has provided morphological evidence for direct physiological modulation of dopaminergic neurons by tachykinins through NKR in the rat retina (A17).