Characterization and synaptic connectivity of melanopsin-containing ganglion cells in the primate retina

Melanopsin is a photopigment expressed in retinal ganglion cells, which are intrinsically photosensitive and are also involved in retinal circuits arising from rod and cone photoreceptors. This circuitry, however, is poorly understood. Here, we studied the morphology, distribution and synaptic input to melanopsin-containing ganglion cells in a New World monkey, the common marmoset (Callithrix jacchus). The dendrites of melanopsin-containing cells in marmoset stratify either close to the inner nuclear layer (outer stratifying), or close to the ganglion cell layer (inner stratifying). The dendritic fields of outer-stratifying cells tile the retina, with little overlap. However, the dendritic fields of outer-stratifying cells largely overlap with the dendritic fields of inner-stratifying cells. Thus, inner-stratifying and outer-stratifying cells may form functionally independent populations. The synaptic input to melanopsin-containing cells was determined using synaptic markers (antibodies to C-terminal binding protein 2, CtBP2, for presumed bipolar synapses, and antibodies to gephyrin for presumed amacrine synapses). Both outer-stratifying and inner-stratifying cells show colocalized immunoreactive puncta across their entire dendritic tree for both markers. The density of CtBP2 puncta on inner dendrites was about 50% higher than that on outer dendrites. The density of gephyrin puncta was comparable for outer and inner dendrites but higher than the density of CtBP2 puncta. The inner-stratifying cells may receive their input from a type of diffuse bipolar cell (DB6). Our results are consistent with the idea that both outer and inner melanopsin cells receive bipolar and amacrine input across their dendritic tree.     

Melanopsin retinal ganglion cells receive bipolar and amacrine cell synapses

Melanopsin is a novel opsin synthesized in a small subset of retinal ganglion cells. Ganglion cells expressing melanopsin are capable of depolarizing in response to light in the absence of rod or cone input and are thus intrinsically light sensitive. Melanopsin ganglion cells convey information regarding general levels of environmental illumination to the suprachiasmatic nucleus, the intergeniculate leaflet, and the pretectum. Typically, retinal ganglion cells communicate information to central visual structures by receiving input from retinal photoreceptors via bipolar and amacrine cells. Because melanopsin ganglion cells do not require synaptic input to generate light-induced signals, these cells need not receive synapses from other neurons in the retina. In this study, we examined the ultrastructure of melanopsin ganglion cells in the mouse retina to determine the type (if any) of synaptic input these cells receive. Melanopsin immunoreaction product was associated primarily with the plasma membrane of (1) perikarya in the ganglion cell layer, (2) dendritic processes in the inner plexiform layer (IPL), and (3) axons in the optic fiber layer. Melanopsin-immunoreactive dendrites in the inner (ON) region of the IPL were postsynaptic to bipolar and amacrine terminals, whereas melanopsin dendrites stratifying in the outer (OFF) region of the IPL received only amacrine terminals. These observations suggested that rod and/or cone signals may be capable of modifying the intrinsic light response in melanopsin-expressing retinal ganglion cells.     

Synaptic connections of DB3 diffuse bipolar cell axons in macaque retina

In primate retinas, the dendrites of DB3 diffuse bipolar cells are known to receive inputs from cones. The goal of this study was to describe the synaptic connections of DB3 bipolar cell axons in the inner plexiform layer. DB3 bipolar cells in midperipheral retina were labeled with antibodies to calbindin, and their axons were analyzed in serial, ultrathin sections by electron microscopy. Synapses were found almost exclusively at the axonal varicosities of DB3 axon terminals. There were 2.14 synaptic ribbons per varicosity. There were 33 varicosities per DB3 cell, giving an average of 71 ribbons per axon terminal. Because there were 1.5 postsynaptic ganglion cell dendrites per DB3 axonal varicosity, we estimate that there is at least 1 synapse per varicosity onto a parasol ganglion cell dendrite. There were 3.4 input synapses from amacrine cells per axonal varicosity. Among these were feedback synapses to the DB3 bipolar cell axon varicosities, which were made by 47% of the postsynaptic amacrine cell processes. Some of the feedback synapses could be from amacrine cells immunoreactive for cholecystokinin precursor or choline acetyltransferase, because both types of amacrine cells costratify with parasol cells and are known to be presynaptic to bipolar cells. AII amacrine cells were both presynaptic and postsynaptic to DB3 axons, a finding consistent with the large rod input to parasol ganglion cells reported in physiological experiments. DB3 bipolar cell axons also made frequent contacts with neighboring DB3 axons, and gap junctions were always found at these sites.     

Light and electron microscopy of the ground squirrel retina: Functional considerations

Light and electron microscopy of Golgi-impregnated ground squirrel retinas have revealed a range of morphological subtypes of bipolar, amacrine, and ganglion cells. There are at least seven subtypes of bipolar cells. Those subtypes in which the somata were high (sclerad) in the inner nuclear layer (3 subtypes) had axon terminals low (vitread) in the inner plexiform layer, and those with somata low in the inner nuclear layer (4 subtypes) had axon terminals high in the inner plexiform layer. The bipolar subtypes with high axon terminals made flat contacts with receptor cells, whereas all but one of the bipolar subtypes with low axon terminals made ribbon-related contacts with receptor cells. There are at least five subtypes of amacrine cells. The two subtypes which the Golgi method revealed most frequently were a broad-field, unistratified neuron with a dendritic spread in excess of 1,000 m?m and a narrow-field, diffuse neuron with a dendritic spread of about 30 m?m. The broad-field, unistratified cell had the lowest proportion of amacrine vs. bipolar cell synaptic input of the amacrine subtypes (43%), whereas the narrow-field, diffuse cell had one of the greatest proportions of amacrine cell input (96%). There are at least 15 subtypes of ganglion cells. The proportion of synaptic inputs to these cells ranged from 21% to 100% amacrine cell synapses. An attempt has been made to relate this new knowledge of retinal circuitry to the physiological output of the ganglion cells.     

Midget ganglion cells of the parafovea of the human retina: a study by electron microscopy and serial section reconstructions.

In this study we used serial section electron microscopy and three-dimensional reconstructions to examine four midget ganglion cells of the human retina. The four cells were located in the parafoveal retina 2.5 mm or 8 degrees from the foveal center. Both type a (with dendritic trees in distal inner plexiform layer) and type b (with dendritic trees in proximal inner plexiform layer) midget ganglion cells have been studied. These cells have dendritic trees of 7-9 microns diameter, and their complete dendritic trees in the neuropil of the inner plexiform layer can be analyzed, as well as the bipolar cell axon terminals having synaptic input, by a study of 100-150 serial ultrathin sections. Type a midget ganglion cells appear to be in a one-to-one relationship with flat midget bipolar cell axon terminals ending in distal inner plexiform layer. Type b midget ganglion cells are in a one-to-one synaptic relationship with invaginating midget bipolar cell axon terminals in proximal inner plexiform layer. The midget bipolar cells primarily involved with the midget ganglion cells do not contact other ganglion cell dendrites. In other words, midget bipolar cells appear to be in exclusive contact with single midget ganglion cells in the human retina. The midget ganglion cells receive most of their input from their associated midget bipolar cells in the form of ribbon synapses at dyads or monads (55-81 ribbons total), although ribbonless synapses are seen occasionally. In all four midget ganglion cells reconstructed, one or two other bipolar cell axon terminals, presumed to be from wide-field bipolar types, provide 1-3 ribbon synapses each. The number of amacrine synapses upon a midget ganglion cell's dendritic tree is approximately equal to the number of bipolar ribbon inputs (43%-56% bipolar ribbons: 44%-57% amacrine synapses). We assume from our knowledge of response characteristics of ganglion cells in other mammalian retinas (Nelson et al., '78: J. Neurophysiol. 41:427-483), that the type a midget ganglion cell and its exclusive connectivity with a flat midget bipolar cell forms a single cone connected OFF-center pathway, whereas the type b midget ganglion cell with its exclusive connectivity to an invaginating midget bipolar cell forms a single cone connected ON-center pathway, through the retina to the brain.     

Tyrosine hydroxylase immunoreactivity in the rhesus monkey retina reveals synapses from bipolar cells to dopaminergic amacrine cells

The synaptic organization of dopamine-containing amacrine cells in the rhesus monkey retina was studied using immunohistochemistry of tyrosine hydroxylase (TH), the rate-limiting enzyme in the catecholamine synthetic pathway. Cell bodies of the TH-containing neurons were primarily in the innermost tier of the inner nuclear layer. Their synaptic processes, confined to the outermost stratum of the inner plexiform layer, contained mostly small, clear vesicles and were presynaptic to unlabeled amacrine cell processes and cell bodies at junctions that were symmetrical. Synapses onto the TH-immunoreactive neurons were from bipolar cell axon terminals, nonimmunoreactive amacrine cell processes, and other TH-containing amacrine cells in a decreasing order of predominance. The bipolar cells were presynaptic to the TH-containing neuronal processes at ribbon synapses. The size, structure, and position of the bipolar cell axon terminals, which, like the TH-reactive processes, were narrowly confined to the outermost stratum of the inner plexiform layer, indicate that they are recently described giant bistratified bipolar cells. The identification of this bipolar cell input now provides evidence for a pathway from the outer plexiform layer to dopaminergic amacrine cells in the inner plexiform layer via a type of cone bipolar cell.     

Electron microscopic analysis of amacrine and bipolar cell inputs on Y-, X- and W-cells in the cat retina

In the cat retina, bipolar and amacrine cell inputs were analyzed electron microscopically in 5 ganglion cells (two Y-cells, two X-cells and one W-cell) that were well-isolated and had clear morphological features. For Y- and X-cells, subtypes of a and b were further identified according to the sublamina of the inner plexiform layer in which their dendrites extended. Y-a and Y-b ganglion cells had large somas, thick axons, and several thick dendrites that branched extensively with a large dendritic field. X-a and X-b cells had medium-sized somas, medium-sized axons and extremely narrow dendritic fields. The W-cell studied had a medium-sized soma, a medium-sized axon, and extremely thin dendrites that extended widely. For each of the 5 ganglion cells, ultrathin serial sections were made to study relative occurrence of amacrine and bipolar synapses in whole length of dendrites. About 50% of the terminals were bipolar in the Y-a and Y-b cell dendrites, 36-38% in the X-a and X-b cell dendrites, whereas only 19.7% were bipolar in the W cell dendrites. Bipolar terminals tended to make synaptic contacts with the distal dendrites of Y- and W-cells.     

Synaptic inputs to physiologically identified retinal X-cells in the cat.

The cat retina contains a number of different classes of ganglion cells, each of which has a unique set of receptive field properties. The mechanisms that underlie the functional differences among classes, however, are not well understood. All of the afferent input to retinal ganglion cells are from bipolar and amacrine cell terminals in the inner plexiform layer, suggesting that the physiological differences among cat retinal ganglion cells might be due to differences in the proportion of input that they receive from these cell types. In this study, we have combined in vivo intracellular recording and labeling with subsequent ultrastructural analysis to determine directly the patterns of synaptic input to physiologically identified X-cells in the cat retina. Our primary aim in these analyses was to determine whether retinal X-cells receive a characteristic pattern of bipolar and amacrine cell input, and further, whether the functional properties of this cell type can be related to identifiable patterns of synaptic input in the inner plexiform layer. We reconstructed the entire dendritic arbor of an OFF-center X-cell and greater than 75% of the dendritic tree of an ON-center X-cell and found that 1) both ON- and OFF-center X-cells are contacted with approximately the same frequency by bipolar and amacrine cell terminals, 2) each of these input types is distributed widely over their dendritic fields, and 3) there is no significant difference in the pattern of distribution of bipolar and amacrine cell synapses onto the dendrites of either cell type. Comparisons of the inputs to the ON- and the OFF-center cell, however, did reveal differences in the complexity of the synaptic arrangements found in association with the two neurons; a number of complex synaptic arrangements, including serial amacrine cell synapses, were found exclusively in association with the dendrites of the OFF-center X-cell. Most models of retinal X-cell receptive fields, because their visual responses share a number of features with those of bipolar cells, have attributed X-cell receptive field properties to their bipolar cell inputs. The data presented here, the first obtained from analyzing the inputs to the entire dendritic arbors of retinal X-cells, demonstrate that these retinal ganglion cells receive nearly one-half of their input from amacrine cells. These results clearly indicate that further data concerning the functional consequences of amacrine cell input are needed to understand more fully visual processing in the X-cell pathway.     

Synaptic connectivity of the diffuse bipolar cell type DB6 in the inner plexiform layer of primate retina

Diffuse bipolar cells in primate retina receive synaptic input from multiple cones and provide output to ganglion cells. Diffuse bipolar cells can be subdivided into six types (DB1-DB6) according to the stratification of their axon terminals in the inner plexiform layer, but their synaptic connectivity in the inner plexiform layer is not well understood. Here the stratification and synaptic connectivity of DB6 axon terminals were studied in the retinae of New World (marmoset) and Old World (macaque) monkeys. Immunohistochemical markers were applied to retinal sections. The sections were analyzed by confocal and deconvolution light microscopy as well as electron microscopy. The DB6 cells were identified with antibodies against CD15; rod bipolar cells were identified with antibodies against protein kinase Calpha (PKCalpha); and AII amacrine cells were identified with antibodies against calretinin. The axons of DB6 and rod bipolar cells occupy distinct regions in stratum 5 of the inner plexiform layer. The distal processes of calretinin-labeled AII cells are usually closely associated with rod bipolar axons but sometimes also with DB6 axons. Pre-embedding immunoelectron microscopy showed that the vast majority (over 86%) of the synaptic output of DB6 cells is onto amacrine cell processes, whereas less than 14% goes to ganglion cell processes. In double-labeled preparations DB6 axons occasionally made output onto calretinin-labeled amacrine processes. Thus it is possible that AII cells receive some input from DB6 cells.     

Synaptic Contacts and the Transient Dendritic Spines of Developing Retinal Ganglion Cells

The dendrites of ganglion cells in the mammalian retina become extensively remodelled during synapse formation in the inner plexiform layer. In particular, after birth in the cat, many short spiny protrusions are lost from the dendrites of ganglion cells during the time when ribbon, presumably bipolar, synapses appear in the inner plexiform layer and when conventional, presumed amacrine, synapses increase significantly in number. It has therefore been postulated that these transient spines may be the initial or preferred substrates for competitive interactions between amacrine or bipolar cell terminals that subsequently result in the formation of appropriate synapses onto the ganglion cells. If so, the majority of synapses made onto developing ganglion cells should be found on these dendritic spines. To test this hypothesis, we determined the synaptic connectivity of identified ganglion cells in the postnatal cat retina during the period of peak spine loss and synapse formation. The dendritic trees of ganglion cells were intracellularly filled with Lucifer yellow that was subsequently photo-oxidized into an electron-dense product suitable for electron microscopy. In serial reconstructions of the dendrites of a postnatal day 11 (P11) alpha ganglion cell and a P14 beta ganglion cell, conventional and ribbon synapses were found predominantly on dendritic shafts. Only three out of a total of 341 dendritic spines from the two cells received direct presynaptic input, all of which were conventional synapses. Thus, our observations suggest that the transient dendritic spines are not the preferred postsynaptic sites as previously suspected. However, it is possible that these structures play a different role in synaptogenesis, such as mediating interactions between retinal neurons that may lead to cell-cell recognition, a necessary step prior to synapse formation at the appropriate target sites (Cooper and Smith, Soc. Neurosci. Abstr. , 14 , 893, 1988).     

Immunohistochemical identification and synaptic inputs to the diffuse bipolar cell type DB1 in macaque retina

Detailed analysis of the synaptic inputs to the primate DB1 bipolar cell has been precluded by the absence of a suitable immunohistochemical marker. Here we demonstrate that antibodies for the EF-hand calcium-binding protein, secretagogin, strongly label the DB1 bipolar cell as well as a mixed population of GABAergic amacrine cells in the macaque retina. Using secretagogin as a marker, we show that the DB1 bipolar makes synaptic contact with both L/M as well as S-cone photoreceptors and only minimal contact with rod photoreceptors. Electron microscopy showed that the DB1 bipolar makes flat contacts at both triad-associated and nontriad-associated positions on the cone pedicle. Double labeling with various glutamate receptor subunit antibodies failed to conclusively determine the subunit composition of the glutamate receptors on DB1 bipolar cells. In the IPL, DB1 bipolar cell axon terminals expressed the glycine receptor, GlyRα1, at sites of contact with AII amacrine cells, suggesting that these cells receive input from the rod pathway.     

Enigmatic bipolar cell of rat visual cortex

Our earlier Golgi-electron microscopic study of bipolar cells in the rat visual cortex showed the axons of these neurons as forming asymmetric synapses (Peters and Kimerer; J. Neurocytol, 10:921-946, '81) in which the most common postsynaptic elements were dendritic spines. This result was unexpected, since Parnavelas (Parnavelas, Sullivan, Lieberman, and Webster: Cell Tissue Res. 183:499-517, '77) had earlier shown a bipolar cell from the same cortex to have an axon forming symmetric synapses with dendritic shafts. Here then was an enigma, strengthened by examination of neuronal components labelled by antibodies to two compounds in particular--namely, vasoactive intestinal polypeptide (VIP) and choline acetyltransferase (ChAT). Antibodies to these compounds preferentially label bipolar cells in the rat cerebral cortex, and the labelled axon terminals form symmetric synapses. Against this background the present study was performed, and it has been shown that the resolution to the enigma is that there are two different populations of bipolar cells in the rat visual cortex. Thus some Golgi-impregnated bipolar cells examined by electron microscopy after gold toning have been found to possess axons forming asymmetric synapses, and others have been found to have axons forming symmetric synapses. The axons of the bipolar cells forming asymmetric synapses most commonly synapse with dendritic spines (67%), although other terminals synapse with dendritic shafts (33%). In contrast, the bipolar cells with axons forming symmetric synapses preferentially synapse with dendritic shafts (100%). The population of bipolar cells that form symmetric synapses includes the ones that label with antibodies to vasoactive intestinal polypeptide (VIP), for the axons of VIP-labelled bipolar cells have been traced to labelled terminals forming symmetric synapses. However, examination of the population of VIP-labelled axon terminals shows that in addition to dendritic shafts, some of the labelled terminals synapse with the cell bodies of pyramidal and nonpyramidal cells. This includes bipolar cells, some of which receive large numbers of VIP-labelled axon terminals. It is also shown that some VIP-positive bipolar cells have myelinated axons. Analysis of tissue labelled with VIP antibody reveals that about 50% of the total population of bipolar cells in the rat visual cortex is VIP positive. These results are discussed in the light of information about labelling of bipolar cells with antibodies to gamma-aminobutyric acid (GABA) and to other peptides, and it is suggested that most VIP-positive bipolar cells also contain GABA.     

Synapses of cone horizontal cell axons in goldfish retina

The axon terminals of cone horizontal cells in the goldfish retina form typical chemical synaptic contacts in the middle of the inner nuclear layer. Approximately 60% of the identified postsynaptic elements were perikarya, axons and dendrites of bipolar cells. The other identified postsynaptic elements were perikarya and processes of interplexiform cells. We propose that the horizontal cell axon terminal contribute to the antagonistic surround responses of the bipolar cells and that they modulate inputs to the outer plexiform layer conveyed by interplexiform cells. Output synapses from horizontal cell axons to unidentified neuronal processes as well as occasional input synapses to the axons from interplexiform cell processes and unidentified perikarya were also observed in the same region of the inner nuclear layer.     

Synapses from bipolar cells onto dopaminergic amacrine cells in cat and rabbit retinas

Dopaminergic amacrine cells in the vertebrate retina have long been characterized as 'interamacrine' as they were only found to be pre- and postsynaptic to other amacrine cells. Immunohistochemistry with antibodies directed against tyrosine hydroxylase (TH) revealed synapses from bipolar cell axon terminals to TH-containing neuronal processes at ribbon synapses in the rhesus monkey retina. This finding challenged the notion of the dopaminergic amacrine cell phenotype as 'interamacrine'. In order to determine if the finding of synapses from bipolar cells to dopaminergic amacrine cells could be generalized to other species, we studied the synaptic organization of dopaminergic amacrine cells in the retinas of cats and rabbits with electron microscopy of TH immunoreactivity. In both species, TH-immunoreactive processes were found to be postsynaptic to bipolar axon terminals at ribbon synapses demonstrating that the original finding in the primate may be a significant feature in the retinas of many other vertebrates as well.     

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)     

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.     

Electron microscopical observations on the indoleamine-accumulating neurons and their synaptic connections in the retina of the cat

The distribution of indoleamine-accumulating amacrine cells and their synaptic connections in the retina of the cat were analyzed in the fluorescence, phase-contrast, and electron microscopes. The findings were compared to recently characterized morphological subclasses of amacrine cells. The indoleamine-accumulating neurons were visualized after labeling with an exogenous indoleamine, 5, 6-dihydroxytryptamine. The intravitreal injection of the labeling drug was preceded by treatment with the neurotoxic dopamine-analogue, 6-hydroxydopamine, in order to destroy the otherwise interfering dopaminergic processes. The analysis in the fluorescence and phase-contrast microscopes confirmed earlier reports that the indoleamine-accumulating cell bodies and processes have a distribution consistent with that of amacrine cells. A stratified branching pattern of the indoleamine-accumulating processes in the outer half of the inner plexiform layer was discovered. In the inner half of that layer the branching pattern is diffuse. In the electron microscope the indoleamine-accumulating neurons were seen to have synapses of the conventional type. Their main synaptic contacts are reciprocal synapses with rod bipolar terminals in sublamina b of the inner plexiform layer. They also have synapses with flat cone bipolar terminals in sublamina a, and occasionally with amacrine cells and ganglion cells throughout the inner plexiform layer. Synapses with invaginating cone bipolar terminals, in sublamina b, appear to be rare. The synaptic arrangement with reciprocal synapses with rod bipolar terminals is similar to that of the recently reported AI amacrine cells. It is also similar to that of the indoleamine-accumulating neurons in the retinae of other mammals investigated earlier.     

Serotonergic and serotonin-accumulating neurons in the goldfish retina

Autoradiography of goldfish retinas incubated in micromolar levels of 3H-serotonin displayed 3 kinds of labeled somas in the inner nuclear layer: S1 amacrine cells with heavy labeling, large somas, and a sparse distribution (approximately 93/mm2); S2 amacrine cells with moderate labeling, smaller somas, and a denser distribution (approximately 500/mm2); and a subset of bipolar cells with light labeling, small somas, and a very dense distribution (approximately 4000/mm2). Serotonin-like immunoreactivity was observed only in S1 amacrine cells and their synaptic terminals. Radiolabeled terminals in the inner plexiform layer formed 4 strata that were differentially assigned to the 3 cell types. S1 amacrine cells arborized in sublayers 1 and 5, received inputs from type a1 bipolar cells and amacrine cells, and made synapses on other amacrine cells, type a1 bipolar cells and unidentified processes. Thus, S1 amacrine cells seem to receive significant input from "off-center" pathways. S2 amacrine cells arborized in sublayer 3 and made synapses onto amacrine cells. Labeled bipolar cell terminals were exclusively located in sublayer 2 and were identified as type a2 mixed rod-cone bipolar cells. We conclude that the S1 amacrine cell is truly serotonergic and that radiolabeling of S2 amacrine cells and type a2 bipolar cells is due to cross-specificity for another carrier or processes unrelated to their neurochemical identities. These observations partially reconcile many previous observations on the types, numbers, and synaptologies of teleost retinal neurons identified by different markers for indoleaminergic transmission.     

Observations on the synaptic organization of the retina of the albino rat: a light and electron microscopic study

An electron microscopic study of the retina of the albino rat, with particular emphasis on the synaptic organization of the inner and outer plexiform layers, has been correlated with specimens impregnated with a modified Golgi technique. The central element of the photoreceptor “triad” in the outer plexiform layer is a bipolar cell dendrite. Two types of synaptic contacts were observed in the inner plexiform layer, the “dyad” ribbon synapse and the conventional synapse. The postsynaptic elements of the “dyad” consisted of an amacrine process and a ganglion cell dendrite. Conventional synapses were made by amacrine processes which were usually presynaptic to bipolar terminals. Reciprocal synapses between processes making ribbon synapses and those making conventional synapses were seen. Golgi technique revealed the presence of two types of bipolar cells, three types of amacrine cells, and one type each of horizontal and ganglion cell. These findings are discussed in relation to reported receptive field organization.     

Synaptic inputs to two types of koniocellular pathway ganglion cells in marmoset retina

The retinal connectivity of the diverse group of cells contributing to koniocellular visual pathways (widefield ganglion cells) is largely unexplored. Here we examined the synaptic inputs onto two koniocellular-projecting ganglion cell types named large sparse and broad thorny cells. Ganglion cells were labeled by retrograde tracer injections targeted to koniocellular layer K3 in the lateral geniculate nucleus in marmosets (Callithrix jacchus) and subsequently photofilled. Retinal preparations were processed with antibodies against the C-terminal binding protein 2, the AMPA receptor subunit GluR4, and against CD15 to identify bipolar (excitatory) and/or antibodies against gephyrin to identify amacrine (inhibitory) input. Large sparse cells are narrowly stratified close to the ganglion cell layer. Broad thorny ganglion cells are broadly stratified in the center of the inner plexiform layer. Bipolar input to large sparse cells derives from DB6 and maybe other ON bipolar types, whereas that to broad thorny cells derives from ON and OFF bipolar cell types. The total number of putative synapses on broad thorny cells is higher than the number on large sparse cells but the density of inputs (between 2 and 5 synapses per 100 μm(2) dendritic area) is similar for the two cell types, indicating that the larger number of synapses on broad thorny cells is attributable to the larger membrane surface area of this cell type. Synaptic input density is comparable to previous values for midget-parvocellular and parasol-magnocellular pathway cells. This suggests functional differences between koniocellular, parvocellular, and magnocellular pathways do not arise from variation in synaptic input densities.