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.     

Electron microscopic analysis of the rod pathway of the rat retina

Two immunocytochemical markers were used to label the rod pathway of the rat retina. Rod bipolar cells were stained with antibodies against protein kinase C and AII-amacrine cells with antibodies against parvalbumin. The synaptic circuitry of rod bipolars in the inner plexiform layer (IPL) was studied. Rod bipolar cells make approximately 15 ribbon synapses (dyads) in the IPL. Both postsynaptic members of the dyads are amacrine cells; one is usually the process of an AII-amacrine cell and the other one frequently provides a reciprocal synapse. No direct output from rod bipolar cells into ganglion cells was found. AII-amacrine cells make chemical output synapses with cone bipolar cells and ganglion cells in sublamina a of the IPL. They make gap junctions with cone bipolar cells and other AII-amacrine cells in sublamina b of the IPL. The rod pathway of the rat retina is practically identical to that of the cat and of the rabbit retina. It is very likely that this circuitry is a general feature of mammalian retinal organization. © Wiley-Liss, Inc.     

Axonal neurofilament-H immunolabeling in the rabbit retina

A monoclonal antibody directed at the multiphosphorylated epitope of axonal neurofilament-H (NF-H) was used to label axon-like fibers in the rabbit retina. NF-H-immunopositive fibers were found in the outer plexiform layer (OPL), inner plexiform layer (IPL), and optic fiber layer (OFL). The morphological characteristics of the labeled processes identified those in the OPL as horizontal cell axons and axon terminals and fibers in the OFL as axons of ganglion cells. The NF-H-positive profiles in the OPL formed a subset of horizontal cell processes labeled for calbindin. In the IPL, NF-H-immunoreactive profiles lay at all levels but were detected most often in the middle strata, 2-4. Occasionally, we observed NF-H-immuoreactive processes emerging from the IPL and entering either the GCL or the inner nuclear layer (INL). The labeled fibers in the IPL were typically very thin, less than 1 microm in diameter, and could often be followed for over 1 mm as they ran laterally across the retina. Cell bodies were never labeled by the immunoserum. To identify the NF-H-immunopositive fibers in the IPL, standard immunocytochemical double-labeling techniques were applied, using antibodies directed against several neurotransmitters or modulators thought to be expressed by axon-bearing amacrine cells. The NF-H-positive processes in the IPL were found to correspond to those labeled for tyrosine hydroxylase, somatostatin, substance P, and NADPH diaphorase activity. However, the NF-H labels did not colocalize with those against the vasoactive intestinal peptide-associated protein PHM27. Our results indicate that putative axons in the retina possess the multiphosphorylated NF-H protein found within classic axons in the central nervous system. These results thus support the idea that certain subtypes of amacrine and horizontal cells maintain true axons in the mammalian retina.     

Morphology of bipolar cells labeled by DAPI in the rabbit retina.

The morphology, distribution, and coverage of certain cone bipolar cell types were investigated in rabbit retina. Brief in vitro incubation of isolated rabbit retina in the fluorescent dye 4,6-diamino-2-phenylindole labeled only a few cell types in the inner nuclear layer. Intracellular injection of Lucifer Yellow into these types showed them to be horizontal cells and cone bipolar cells. All stained bipolar cells ramified in sublamina a of the inner plexiform layer (IPL) and formed three classes. Two types ranged from 20 to 60 microns in diameter in both plexiform layers; the other large bipolar cell was 40-70 microns in diameter in the outer plexiform layer (OPL) and up to 150 microns in diameter in the IPL. The brightest type was narrowly stratified in the outer portion of sublamina a. Its density increased from about 500 cells/mm2 in the periphery to about 2,500 cells/mm2 in the visual streak. Staining of neighboring cells of this type showed that processes in the IPL rarely crossed, but often converged at a common site so as to impart a "honeycomb" appearance to a single sublayer of retina. The other small bipolar cell was similar in density and coverage, but stratified diffusely throughout sublamina a. The large bipolar cell stratified narrowly in the distal portion of sublamina a and was more sparsely distributed. Whether determined by staining adjacent cells or by density vs. area calculations, coverage in the OPL approached 1 for each type, as did coverage in the IPL for the two types with narrow fields.     

‘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.     

Dependence of cytochrome oxidase activity in the rat lateral geniculate nucleus on retinal innervation

Patterns of cytochrome oxidase (CO) activity were examined histochemically in the dorsal lateral geniculate nucleus (LGNd) and retina of pigmented rats. CO staining was not uniform and was distributed in a pattern similar to that of retinal afferents. Portions of the LGNd receiving an exclusively crossed projection were moderately reactive whereas regions receiving an uncrossed or overlapping crossed and uncrossed projection were darkly reactive. The dependence of oxidative metabolic activity in the LGNd on retinal innervation was verified in animals with unilateral enucleation. In adults, chronic monocular enucleation led to a decrease in CO staining in portions of the LGNd deprived of retinal input; in animals enucleated at birth, normal patterns of CO reactivity failed to develop and both LGNds had a more uniform pattern of moderate CO staining. Most neurons in the ganglion cell layer of the retina were moderately reactive for CO. However, there were approximately 3,000 darkly reactive cells, most of which appear to be ganglion cells. The darkly reactive cells were more numerous in the peripheral temporal retina. The laminar pattern of CO staining in the retina was similar to that described previously for carnivores and primates. The most reactive laminae were the inner and outer plexiform layers and the photoreceptor inner segments. Within the inner plexiform layer, sublamina a was more darkly stained than sublamina b. These results suggest that the physiological properties of crossed and uncrossed visual pathways in rats are functionally dissimilar at the level of both the retina and the LGNd.     

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.     

Regional specialization of the rat retina: catecholamine-containing amacrine cell characterization and distribution

The distribution of catecholaminergic amacrine cells has been investigated in rats by means of immunohistochemical labelling of wholemounted retinas. Two groups of catecholamine-containing cells could be distinguished on the basis of their catecholamine and biosynthetic enzyme content. Both groups could be stained with an anti-tyrosine hydroxylase (TH) antiserum. The first group was composed of large, strongly TH-immunoreactive stellate amacrine cells, located principally in the innermost row of the inner nuclear layer (INL) and sending processes to the outermost sublamina of the inner plexiform layer (IPL). Some were displaced in the IPL or in the ganglion cell layer (GCL). This first group of cells can be regarded as dopaminergic since they were also stained by an anti-dopamine (DA) antiserum. The second group was composed of small, weakly TH-positive cell bodies, located slightly more sclerad within the INL. Their processes were usually not labelled with anti-TH. Identical cells could be better visualized with an anti-phenylethanolamine-N-methyltransferase (PNMT) antiserum. Their processes were observed in the middle sublamina of the IPL. A great number of these cells were displaced in the GCL. They could be regarded as epinephrine cells. Concerning the density and distribution throughout the retina a striking difference was observed between the superior and inferior halves of the retina, whereas a lower difference was observed between the nasal and temporal regions. Almost all the PNMT-immunoreactive cells were located throughout the upper retina, whereas the DA-cells were especially concentrated in the upper temporal quadrant. The distribution of the DA cells parallels that of the ganglion cells whose density is also maximal in the upper temporal retina.     

NADPH-diaphorase neurons in the retina of the hamster

NADPH-diaphorase-positive neurons have been demonstrated in the inner nuclear layer and ganglion cell layer of the retina of different mammalian species, but so far no experiments have been conducted to identify whether these cells are amacrine cells and/or retinal ganglion cells. We attempted to solve this problem by studying the NADPH-diaphorase-positive neurons in the hamster retina. From the NADPH-diaphorase histochemical reaction, two distinct types of neurons in the hamster retina were identified. They were named ND(g) and ND(i) cells. The ND(g) cells were cells with larger somata, ranging from 10 to 21 μm in diameter with a mean of 15.58 μm (S.D.= 2.59). They were found in the ganglion cell layer only. The ND(i) cells were smaller, with the somata ranging from 7 to 11 μm and having the mean diameter of 8.77 μm (S.D. = 1.24). Most of the ND(i) cells were found in the inner nuclear layer, and only very few could be observed in the inner plexiform layer. On average, there were 8,033 ND(g) and 5,051 ND(i) cells in the ganglion cell layer and inner nuclear layer, respectively. Two experiments were performed to clarify whether any of the NADPH-diaphorase neurons were retinal ganglion cells. Following unilateral optic nerve section, which leads to the retrograde degeneration of retinal ganglion cells, the numbers of both ND(g) and ND(i) cells did not change significantly for up to 4 months. In addition, when retinal ganglion cells were prelabeled retrogradely (horseradish peroxidase of flurescent microspheres) and retinas were then stained for NADPH diaphorase, no double-labeled neurons were detected. These results indicated that the NADPH-diaphorase neurons in the hamster retina were the amacrine cells in the inner nuclear layer and displaced amacrine cells in the ganglion cell layer. Dendrites of the ND(g) and ND(i) cells were found to stratify in sublaminae 1, 3, and 5 of the inner plexiform layer, with a prominent staining in the sublamina 5. The possible importance of this arrangement in the rod pathway is also discussed. © 1994 Wiley-Liss, Inc.     

Localization of GABA, glycine, glutamate and tyrosine hydroxylase in the human retina.

A light microscope study using postembedding immunocytochemistry techniques to demonstrate the common neurotransmitter candidates gamma-aminobutyric acid (GABA), glycine, glutamate, and tyrosine hydroxylase for dopamine has been done on human retina. By using an antiserum to GABA, we found GABA-immunoreactivity (GABA-IR) to be primarily in amacrine cells lying in the inner nuclear layer (INL) or displaced to the ganglion cell layer (GCL). A few stained cells in the INL, which are probably interplexiform cells, were observed to project thin processes towards the outer plexiform layer (OPL). There were heavily stained bands of immunoreactivity in strata 1, 3 and 5 of the inner plexiform layer (IPL). An occasional ganglion cell was also GABA-IR. By using an antiserum to glycine, stained cells were observed at all levels of the INL. Most of these were amacrines, but a few bipolar cells were also glycine-IR. Displaced amacrine cells and large-bodied cells, which are probably ganglion cells, stained in the GCL. The bipolar cells that stained appeared to include both diffuse and midget varieties. The AII amacrine cell of the rod pathway was clearly stained in our material but at a lower intensity than two other amacrine cell types tentatively identified as A8 and A3 or A4. Again, there was stratified staining in the IPL, with strata 2 and 4 being most immunoreactive. An antiserum to glutamate revealed that most of the neurons of the vertical pathways in the human retina were glutamate-IR. Rod and cone photoreceptor synaptic endings labeled as did the majority of bipolar and ganglion cells. The rod photoreceptor stained more heavily than the cone photoreceptor in our material. While both midget and diffuse cone bipolar cell types were clearly glutamate-IR, rod bipolars were not noticeably stained. The most strongly staining glutamate-IR processes of the IPL lay in the outer half, in sublamina a. The antiserum to tyrosine hydroxylase (TOH) revealed two different amacrine cell types. Strongly immunoreactive cells (TOH1) had their cell bodies in the INL and their dendrites ramified in a dense plexus in stratum 1 of the IPL. Fine processes arising from their cell bodies or from the stratum 1 plexus passed through the INL to reach the OPL but did not produce long-ranging ramifications therein. The less immunoreactive amacrines (TOH2) lay in the INL, the center of the IPL or the GCL and emitted thick dendrites that were monostratified in stratum 3 of the IPL.     

Lack of early pattern stimulation prevents normal development of the alpha (Y) retinal ganglion cell population in the cat

Binocular deprivation of pattern vision (BD) early in life permanently impairs global motion perception. With the SMI-32 antibody against neurofilament protein (NFP) as a marker of the motion-sensitive Y-cell pathway (Van der Gucht et al. [2001] Cereb. Cortex 17:2805-2819), we analyzed the impact of early BD on the retinal circuitry in adult, perceptually characterized cats (Burnat et al. [2005] Neuroreport 16:751-754). In controls, large retinal ganglion cells exhibited a strong NFP signal in the soma and in the proximal parts of the dendritic arbors. The NFP-immunoreactive dendrites typically branched into sublamina a of the inner plexiform layer (IPL), i.e., the OFF inner plexiform sublamina. In the retina of adult BD cats, however, most of the NFP-immunoreactive ganglion cell dendrites branched throughout the entire IPL. The NFP-immunoreactive cell bodies were less regularly distributed, often appeared in pairs, and had a significantly larger diameter compared with NFP-expressing cells in control retinas. These remarkable differences in the immunoreactivity pattern were typically observed in temporal retina. In conclusion, we show that the anatomical organization typical of premature Y-type retinal ganglion cells persists into adulthood even if normal visual experience follows for years upon an initial 6-month period of BD. Binocular pattern deprivation possibly induces a lifelong OFF functional domination, normally apparent only during development, putting early high-quality vision forward as a premise for proper ON-OFF pathway segregation. These new observations for pattern-deprived animals provide an anatomical basis for the well-described motion perception deficits in congenital cataract patients.     

The histochemical localization of cytochrome oxidase in the retina and lateral geniculate nucleus of the ferret, cat, and monkey, with particular reference to retinal mosaics and ON/OFF-center visual channels

The histochemical localization of cytochrome oxidase within the normal retina and lateral geniculate nucleus (LGN) of cats, ferrets, and monkeys revealed that distinct layers, types of cells, and portions of neurons are more intensely stained than others. The dark staining of photoreceptor inner segments and cone pedicles and the light staining of photoreceptor outer segments, somata, and rod spherules demonstrates that different segments of the same cell may have disparate but distinct levels of oxidative enzyme activity. In tangential sections of retina, regular mosaic arrays were evident for each of several darkly reactive retinal components, such as cone inner segments, cone pedicles, and horizontal cells. In the cat and ferret, regular mosaic arrays were also formed by metabolically distinguishable populations of ganglion cells. Ia and IIa ganglion cells (OFF-; Nelson, R., E. V. Famiglietti, Jr., and H. Kolb (1978) J. Neurophysiol. 41: 472-483) were more darkly reactive than the other classes. The darker staining of sublamina a of the inner plexiform layer (OFF-; Famiglietti, E. V., Jr., and H. Kolb (1976) Science 194: 193-195) in the cat and ferret retina, as well as sublamina A' and A1' of the ferret LGN (OFF-; Stryker, M.P., and K.R. Zahs (1983a) J. Neurosci. 3: 1943-1951) suggest that, under typical rearing conditions, the OFF-channels may be metabolically more active than the ON-channels in these species. In Macaca and Saimiri, darker staining was observed in sublamina b of the inner plexiform layer (ON-; Famiglietti, E.V., Jr., and H. Kolb (1976) Science 194: 193-195) and laminae 1, 2, and 6 of the LGN, implying that, under similar rearing conditions, a different pattern is observed. The dark staining of many large retinal ganglion cells, as well as most of the larger LGN neurons (presumed Y/Y-like), in all species studied is evidence that the Y/Y-like pathway is also highly active.     

NADPH diaphorase cells in the mammalian inner retina

The distribution of the enzyme NADPH diaphorase was examined histochemically in the retina of the rat, rabbit, cat, owl monkey, squirrel monkey, rhesus macaque, and human being. In all species tested the enzyme was concentrated in cells 10-12 microns in diameter at the vitread margin of the inner nuclear layer. The population was sparse (less than 2,000 cells/rat retina). In several species additional minor populations were observed. While a clear function for NADPH diaphorase has yet to be described, an abundance of the enzyme characterizes a similar subpopulation of retinal cells in a wide variety of mammals.     

Type 1 nitrergic (ND1) cells of the rabbit retina: comparison with other axon-bearing amacrine cells

NADPH diaphorase (NADPHd) histochemistry labels two types of nitrergic amacrine cells in the rabbit retina. Both the large ND1 cells and the small ND2 cells stratify in the middle of the inner plexiform layer, and their overlapping processes produce a dense plexus, which makes it difficult to trace the morphology of single cells. The complete morphology of the ND1 amacrine cells has been revealed by injecting Neurobiotin into large round somata in the inner nuclear layer, which resulted in the labelling of amacrine cells whose proximal morphology and stratification matched those of the ND1 cells stained by NADPHd histochemistry. The Neurobiotin-injected ND1 cells showed strong homologous tracer coupling to surrounding ND1 cells, and double-labelling experiments confirmed that these coupled cells showed NADPHd reactivity. The ND1 amacrine cells branch in stratum 3 of the inner plexiform layer, where they produce a sparsely branched dendritic tree of 400-600 microm diameter in ventral peripheral retina. In addition, each cell gives rise to several fine beaded processes, which arise either from a side branch of the dendritic tree or from the tapering of a distal dendrite. These axon-like processes branch successively within the vicinity of the dendritic field before extending, with little or no further branching, for 3-5 mm from the soma in ventral peripheral retina. Consequently, these cells may span one-third of the visual field of each eye, and their spatial extent appears to be greater than that of most other types of axon-bearing amacrine cells injected with Neurobiotin in this study. The morphology and tracer-coupling pattern of the ND1 cells are compared with those of confirmed type 1 catecholaminergic cells, a presumptive type 2 catecholaminergic cell, the type 1 polyaxonal cells, the long-range amacrine cells, a novel type of axon-bearing cell that also branches in stratum 3, and a type of displaced amacrine cell that may correspond to the type 2 polyaxonal cell.     

Ectopic retinal ON bipolar cell synapses in the OFF inner plexiform layer: Contacts with dopaminergic amacrine cells and melanopsin ganglion cells

A key principle of retinal organization is that distinct ON and OFF channels are relayed by separate populations of bipolar cells to different sublaminae of the inner plexiform layer (IPL). ON bipolar cell axons have been thought to synapse exclusively in the inner IPL (the ON sublamina) onto dendrites of ON‐type amacrine and ganglion cells. However, M1 melanopsin‐expressing ganglion cells and dopaminergic amacrine (DA) cells apparently violate this dogma. Both are driven by ON bipolar cells, but their dendrites stratify in the outermost IPL, within the OFF sublamina. Here, in the mouse retina, we show that some ON cone bipolar cells make ribbon synapses in the outermost OFF sublayer, where they costratify with and contact the dendrites of M1 and DA cells. Whole‐cell recording and dye filling in retinal slices indicate that type 6 ON cone bipolars provide some of this ectopic ON channel input. Imaging studies in dissociated bipolar cells show that these ectopic ribbon synapses are capable of vesicular release. There is thus an accessory ON sublayer in the outer IPL. J. Comp. Neurol. 517:226‐244, 2009. © 2009 Wiley‐Liss, Inc.     

Identification and morphological classification of horizontal, bipolar, and amacrine cells within the zebrafish retina

Horizontal, bipolar, and amacrine cells in the zebrafish retina were morphologically characterized using DiOlistic techniques. In this method, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI)-coated microcarriers are shot at high speed onto the surfaces of living retinal slices where the DiI then delineates axons, somata, and dendrites of isolated neurons. Zebrafish retinal somata were 5-10 microm in diameter. Three horizontal cell types (HA-1, HA-2, and HB) were identified; dendritic tree diameters averaged 25-40 microm. HA somata were round. Cells classified as HA-2 were larger than HA-1 cells and possessed an axon. HB somata were flattened, without an axon, although short fusiform structure(s) projected from the soma. Bipolar cells were separated into 17 morphological types. Dendritic trees ranged from 10 to 70 microM. There were six B(on) types with axon boutons only in the ON sublamina of the inner plexiform layer (IPL), and seven B(off) types with axon boutons or branches only in the OFF sublamina. Four types of bistratified bipolar cells displayed boutons in both ON and OFF layers. Amacrine cells occurred in seven types. A(off) cells (three types) were monostratified and ramified in the IPL OFF sublamina. Dendritic fields were 60-150 microM. A(on) pyriform cells (three types) branched in the ON sublamina. Dendritic fields were 50-170 microM. A(diffuse) cells articulated processes in all IPL strata. Dendritic fields were 15-90 microM. These findings are important for studies examining signal processing in zebrafish retina and for understanding changes in function resulting from mutations and perturbations of retinal organization.     

AII amacrine cell population in the rabbit retina: Identification by parvalbumin immunoreactivity

Parvalbumin (PV) is a calcium-binding protein localized to selected neurons in the nervous system, including the retina. This investigation evaluated the distribution of PV immunoreactivity in the rabbit retina using immunohistochemistry with a monoclonal antibody directed to carp PV. In the inner nuclear layer (INL), PV immunoreactivity was present in horizontal and amacrine cells. In the ganglion cell layer, PV immunostaining was confined to somata that are likely to be both displaced amacrine cells and ganglion cells. PV-immunoreactive (IR) amacrine cells were positioned in the proximal INL adjacent to the inner plexiform layer (IPL). These cells usually gave rise to a single primary process, which arborized into two distinct bands in the IPL. In sublamina a, the processes were thin and had large, irregular endings. In sublamina b, multiple processes branched from the primary process and were characterized by varicosities and spines. PV-IR amacrine cell bodies measured from 8 to 10 μm in diameter. Their density was highest in the visual streak and lowest in the periphery of the superior retina. The average number of PV-IR amacrine cells was 464,045 cells per retina (N = 3), and the average regularity index of the PV-IR cell mosaic was 3.23. PV-IR amacrine cells were further characterized by double-label immunofluorescence experiments using antibodies to PV and tyrosine hydroxylase (TH). Varicose TH-IR processes were in close apposition to many PV-IR amacrine cells and often formed “ring structures” around them. Together, these morphological, quantitative, and histochemical observations indicate that PV immunoreactivity in the INL is localized predominantly to AII amacrine cells, and therefore it is a valuable marker for the identification of this cell type. © 1995 Wiley-Liss, Inc.     

NADPH diaphorase activity in the rabbit retina is modulated by glutamatergic pathways

NADPH diaphorase activity in the rabbit retina is modulated by the state of visual adaptation. In this study, we tested possible glutamatergic control of this phenomenon. Rabbits were injected intravitreally with agonists and antagonists of glutamate. After adaptation (3 hours) to either room light or darkness, the rabbits were killed and the retinae were prepared for NADPH diaphorase histochemistry. Kainic acid significantly reduced the number of NADPH diaphorase amacrine cells but augmented NADPH diaphorase activity in horizontal cells in both light- and dark-adapted animals. 6,7-Dinitroquinoxaline-2,3(1H,4H)-dione exerted no effect on amacrine cells but eliminated NADPH diaphorase activity in horizontal cells. 2-Amino-4-phosphono butyric acid did not affect NADPH diaphorase activity in horizontal cells but reduced the degree of staining in the neuronal processes of amacrine cells. MK-801 and N-methyl-D-aspartic acid (NMDA) had no effect on NADPH diaphorase activity in horizontal cells. However, MK-801 reduced staining in the neuronal processes of amacrine cells but not in their cell bodies. NMDA effects were expressed in a significant reduction in the number and size of amacrine cells that were NADPH diaphorase positive. These results indicate that activation of NADPH diaphorase in horizontal cells by darkness is mediated by the activation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainic acid (KA)-type glutamate receptors. The ON pathway in the retina is probably involved in modulation of NADPH diaphorase in the neuronal processes of amacrine cells. Amacrine cells that are NADPH diaphorase positive contain NMDA-type and AMPA/KA-type receptors and are highly susceptible to NMDA and kainic acid toxicity.     

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).     

Localization of neuropeptide-immunoreactive neurons in the human retina.

Light microscopic immunocytochemistry was utilized to localize populations of neurons in the human retina immunoreactive for the following neuroactive peptides: substance P (SP), vasoactive intestinal polypeptide (VIP), somatostatin (SOM) and LANT-6-(H-Lys-Asn-Pro-Tyr-Ile-Leu-OH), a hexapeptide which is identical to the C-terminal half of neurotensin except for the amino acid substitutions Lys/Arg and Asn/Arg. The majority of SP immunoreactive cells were amacrine cells whose pear-shaped or oval cell bodies (about 8 microns in diameter) were situated in the proximal parts of the inner nuclear layer. A small number of SP-stained somas (about 10-15 microns in diameter) were located in the ganglion cell layer and were designated as those of displaced amacrine cells. The SP-immunoreactive processes were distributed in sublamina 1, 3 and 5 with the most dense plexus being found in sublamina 3 of the inner layer. VIP-positive cell bodies (8-9 microns) were oval or pear-shaped and were situated in the innermost cell rows of inner nuclear layer. The majority of fine VIP-immunoreactive processes extended to sublamina 3 with only a few branches distributing in sublamina 1 of the inner plexiform layer. The SOM-stained cell bodies (10-11 microns) were round and were situated in the innermost cell rows of inner nuclear layer. SOM-positive processes were observed in sublamina 1 and 2 of the inner plexiform layer. The LANT-6 immunoreactive cell bodies (12-22 microns) were either oval-, round- or pyriform-shaped and were situated in ganglion cell layer.(ABSTRACT TRUNCATED AT 250 WORDS)