Interactions between enkephalin and GABA in avian retina

In addition to conventional neurotransmitters such as acetylcholine, dopamine, glycine and gamma-aminobutyric acid (GABA), a number of peptide-immunoreactive substances have recently been localized in the vertebrate retina. The functional roles of these retinal peptides and their interactions with conventional neurotransmitters are largely unknown. We have previously shown that exogenous opiates affect both the release of GABA and the firing patterns of ganglion cells in the goldfish retina, and we have now begun a systematic characterization of the opioid pathways in the chicken retina, because, among vertebrate retinas, avian retinas contain the highest concentration of enkephalins. Monoclonal antibodies specific for enkephalin have been used to demonstrate that a subpopulation of enkephalin-containing amacrine cells exists in the chicken retina. This retina also synthesizes Met-enkephalin and releases it on cell depolarization. The enkephalin-induced inhibition of GABA release in goldfish retina led us to examine whether similar interactions occur in chicken, and if so, whether enkephalins and GABA coexist in the same amacrine cells. Our results, presented here, indicate that exogenous enkephalins do indeed inhibit GABA release in the chicken retina. Surprisingly, we found that although some amacrine cells contain both enkephalin and GABA, others contain only one or the other.     

Cortical magnification factor and the ganglion cell density of the primate retina

It has long been contentious whether the large representation of the fovea in the primate visual cortex (V1) indicates a selective magnification of this part of the retina, or whether it merely reflects the density of retinal ganglion cells. The measurement of the retinal ganglion-cell density is complicated by lateral displacements of cells around the fovea and the presence of displaced amacrine cells in the ganglion cell layer. We have now identified displaced amacrine cells by GABA immunohistochemistry and by retrograde degeneration of ganglion cells. By reconstructing the fovea from serial sections, we were able to compare the densities of cones, cone pedicles and ganglion cells; in this way we found that there are more than three ganglion cells per foveal cone. Between the central and the peripheral retina, the ganglion cell density changes by a factor of 1,000-2,000, which is within the range of estimates of the cortical magnification factor. There is therefore no need to postulate a selective magnification of the fovea in the geniculate and/or the visual cortex.     

Two types of orientation-sensitive responses of amacrine cells in the mammalian retina.

Neurons sensitive to the orientation of light stimuli exist throughout the mammalian visual system, suggesting that this spatial feature is a fundamental cue used by the brain to decipher visual information. The most peripheral neurons known to show orientation sensitivity are the retinal ganglion cells. Considerable morphological and pharmacological data suggest that the orientation sensitivity of ganglion cells is formed, at least partly, by the amacrine cells, which are laterally oriented interneurons presynaptic to the ganglion cells in the inner plexiform layer. So far there have been few studies of the responses of amacrine cells to oriented visual stimuli and their role in forming orientation-sensitive responses in the retina remains unclear. Here I report the novel finding of a population of amacrine cells in the rabbit retina which are orientation-sensitive. These amacrine cells can be divided into two subtypes, whose orientation sensitivity is manufactured by two distinct mechanisms. The orientation sensitivity of the first subtype of amacrine cell is formed from the interactions of excitatory, centre-receptive field synaptic inputs and inhibitory inputs of opposite polarity, whereas that for cells of the second subtype seems to be the product of a marked asymmetry in their dendritic arbors.     

Transmembrane semaphorin signalling controls laminar stratification in the mammalian retina

In the vertebrate retina, establishment of precise synaptic connections among distinct retinal neuron cell types is critical for processing visual information and for accurate visual perception. Retinal ganglion cells (RGCs), amacrine cells and bipolar cells establish stereotypic neurite arborization patterns to form functional neural circuits in the inner plexiform layer (IPL), a laminar region that is conventionally divided into five major parallel sublaminae. However, the molecular mechanisms governing distinct retinal subtype targeting to specific sublaminae within the IPL remain to be elucidated. Here we show that the transmembrane semaphorin Sema6A signals through its receptor PlexinA4 (PlexA4) to control lamina-specific neuronal stratification in the mouse retina. Expression analyses demonstrate that Sema6A and PlexA4 proteins are expressed in a complementary fashion in the developing retina: Sema6A in most ON sublaminae and PlexA4 in OFF sublaminae of the IPL. Mice with null mutations in PlexA4 or Sema6A exhibit severe defects in stereotypic lamina-specific neurite arborization of tyrosine hydroxylase (TH)-expressing dopaminergic amacrine cells, intrinsically photosensitive RGCs (ipRGCs) and calbindin-positive cells in the IPL. Sema6A and PlexA4 genetically interact in vivo for the regulation of dopaminergic amacrine cell laminar targeting. Therefore, neuronal targeting to subdivisions of the IPL in the mammalian retina is directed by repulsive transmembrane guidance cues present on neuronal processes.     

Hyperpolarization of fish retinal horizontal cells by kainate and quisqualate

Kainic (KA) and quisqualic (QA) acids have a potent depolarizing action on a variety of neurones of the central nervous system, including retinal horizontal cells. We now report the novel finding that at low concentrations (1-3 microM), these 'excitatory' amino acids hyperpolarize horizontal cells of the fish retina. We show that the hyperpolarizing effects of both KA and QA are reversed by the gamma-aminobutyric acid (GABA) antagonist bicuculline, whereas a second GABA antagonist, picrotoxin, reverses the effects of KA, but not of QA. Neither GABA antagonist influences horizontal cell depolarization by 50 microM KA or 50 microM QA, thus the excitatory (depolarizing and inhibitory (hyperpolarizing) effects of the amino acids involve independent mechanisms. We provide evidence that the hyperpolarizing effects are not mediated by the dopaminergic pathways associated with retinal horizontal cells.     

Segregation of object and background motion in the retina

An important task in vision is to detect objects moving within a stationary scene. During normal viewing this is complicated by the presence of eye movements that continually scan the image across the retina, even during fixation. To detect moving objects, the brain must distinguish local motion within the scene from the global retinal image drift due to fixational eye movements. We have found that this process begins in the retina: a subset of retinal ganglion cells responds to motion in the receptive field centre, but only if the wider surround moves with a different trajectory. This selectivity for differential motion is independent of direction, and can be explained by a model of retinal circuitry that invokes pooling over nonlinear interneurons. The suppression by global image motion is probably mediated by polyaxonal, wide-field amacrine cells with transient responses. We show how a population of ganglion cells selective for differential motion can rapidly flag moving objects, and even segregate multiple moving objects.     

GABA and GAD immunoreactivity of photoreceptor terminals in primate retina

Within the vertebrate retina, two types of photoreceptor cells--the rods and cones--transduce visual signals and convey this information through synapses with bipolar and horizontal cells. Although the neurotransmitter at these first-order synapses has not been identified, electrophysiological studies suggest that it might be excitatory. In the present study, however, we have found photoreceptor terminals in the rhesus monkey retina which are immunoreactive with antibodies to either gamma-aminobutyric acid (GABA) or L-glutamic acid decarboxylase (GAD, an enzyme involved in the synthesis of GABA). In the perifoveal region of the retina, approximately 25% of presynaptic profiles having ultrastructural characteristics of either rod or cone terminals are immunoreactive with one or the other antibody. This evidence for a putatively inhibitory neurotransmitter in photoreceptor terminals challenges present understanding of retinal synaptic function.     

Fast neurotransmitter release triggered by Ca influx through AMPA-type glutamate receptors

Feedback inhibition at reciprocal synapses between A17 amacrine cells and rod bipolar cells (RBCs) shapes light-evoked responses in the retina. Glutamate-mediated excitation of A17 cells elicits GABA (gamma-aminobutyric acid)-mediated inhibitory feedback onto RBCs, but the mechanisms that underlie GABA release from the dendrites of A17 cells are unknown. If, as observed at all other synapses studied, voltage-gated calcium channels (VGCCs) couple membrane depolarization to neurotransmitter release, feedforward excitatory postsynaptic potentials could spread through A17 dendrites to elicit 'surround' feedback inhibitory transmission at neighbouring synapses. Here we show, however, that GABA release from A17 cells in the rat retina does not depend on VGCCs or membrane depolarization. Instead, calcium-permeable AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid) receptors (AMPARs), activated by glutamate released from RBCs, provide the calcium influx necessary to trigger GABA release from A17 cells. The AMPAR-mediated calcium signal is amplified by calcium-induced calcium release (CICR) from intracellular calcium stores. These results describe a fast synapse that operates independently of VGCCs and membrane depolarization and reveal a previously unknown form of feedback inhibition within a neural circuit.     

Glutamate and γ-aminobutyric acid receptors and their characteristics in retina

Glutamate and Ⅱ-aminobutyric acid (GABA) are important neurotransmitters in the retinal neuronal circuitry. Using the whole-cell patch clamp technique and a rapid solution changer, glutamate and GABA receptors in the retina have been extensively investigated. Results indicate that glutamate receptors on horizontal cells may be an AMPA preferring subtype, which predominantly consists of flop splice variants. GABAA and GABAC receptors coexist in bipolar cells, with the latter showing significant desensitization. Kinetics analysis has demonstrated that the activation, deactivation and desensitization of the GABAC receptor-mediated response of these cells are overall slower than those of the GABAA response. Endogenous modulator Zn2 + in the retina has been found to differentially modulate the kinetic characteristics of the GABAC and GABAA responses.     

Long-distance intraretinal connections in birds

Electrophysiological experiments have shown in both birds and mammals that remote parts of the retina, several millimetres apart, interact at the retinal level. The anatomical basis of this is poorly understood, although in mammals some cells in the ganglion cell layer have axons that terminate in the inner plexiform layer several millimetres from the cell body. In birds, the longest previously reported intraretinal connections were from amacrine cells, extending only a few hundred microns. But we here describe very long connections that span almost the entire extent of the retina in chicks and chick embryos. The parent cell bodies are in the inner nuclear layer of the ventral half of the retina, and they project in topographical order onto the dorsal half. They do not project to the brain. They may be involved in selective switching of attention between the upper and lower parts of the visual field, at an unprecedentedly early stage of visual processing.     

MEGF10 and MEGF11 mediate homotypic interactions required for mosaic spacing of retinal neurons

In many parts of the nervous system, neuronal somata display orderly spatial arrangements. In the retina, neurons of numerous individual subtypes form regular arrays called mosaics: they are less likely to be near neighbours of the same subtype than would occur by chance, resulting in 'exclusion zones' that separate them. Mosaic arrangements provide a mechanism to distribute each cell type evenly across the retina, ensuring that all parts of the visual field have access to a full set of processing elements. Remarkably, mosaics are independent of each other: although a neuron of one subtype is unlikely to be adjacent to another of the same subtype, there is no restriction on its spatial relationship to neighbouring neurons of other subtypes. This independence has led to the hypothesis that molecular cues expressed by specific subtypes pattern mosaics by mediating homotypic (within-subtype) short-range repulsive interactions. So far, however, no molecules have been identified that show such activity, so this hypothesis remains untested. Here we demonstrate in mouse that two related transmembrane proteins, MEGF10 and MEGF11, have critical roles in the formation of mosaics by two retinal interneuron subtypes, starburst amacrine cells and horizontal cells. MEGF10 and 11 and their invertebrate relatives Caenorhabditis elegans CED-1 and Drosophila Draper have hitherto been studied primarily as receptors necessary for engulfment of debris following apoptosis or axonal injury. Our results demonstrate that members of this gene family can also serve as subtype-specific ligands that pattern neuronal arrays.     

Development of asymmetric inhibition underlying direction selectivity in the retina

Establishing precise synaptic connections is crucial to the development of functional neural circuits. The direction-selective circuit in the retina relies upon highly selective wiring of inhibitory inputs from starburst amacrine cells (SACs) onto four subtypes of ON-OFF direction-selective ganglion cells (DSGCs), each preferring motion in one of four cardinal directions. It has been reported in rabbit that the SACs on the 'null' sides of DSGCs form functional GABA (γ-aminobutyric acid)-mediated synapses, whereas those on the preferred sides do not. However, it is not known how the asymmetric wiring between SACs and DSGCs is established during development. Here we report that in transgenic mice with cell-type-specific labelling, the synaptic connections from SACs to DSGCs were of equal strength during the first postnatal week, regardless of whether the SAC was located on the preferred or null side of the DSGC. However, by the end of the second postnatal week, the strength of the synapses made from SACs on the null side of a DSGC significantly increased whereas those made from SACs located on the preferred side remained constant. Blocking retinal activity by intraocular injections of muscimol or gabazine during this period did not alter the development of direction selectivity. Hence, the asymmetric inhibition between the SACs and DSGCs is achieved by a developmental program that specifically strengthens the GABA-mediated inputs from SACs located on the null side, in a manner not dependent on neural activity.     

Dscam and Sidekick proteins direct lamina-specific synaptic connections in vertebrate retina

Synaptic circuits in the retina transform visual input gathered by photoreceptors into messages that retinal ganglion cells (RGCs) send to the brain. Processes of retinal interneurons (amacrine and bipolar cells) form synapses on dendrites of RGCs in the inner plexiform layer (IPL). The IPL is divided into at least 10 parallel sublaminae; subsets of interneurons and RGCs arborize and form synapses in just one or a few of them. These lamina-specific circuits determine the visual features to which RGC subtypes respond. Here we show that four closely related immunoglobulin superfamily (IgSF) adhesion molecules--Dscam (Down's syndrome cell adhesion molecule), DscamL (refs 6-9), Sidekick-1 and Sidekick-2 (ref. 10)--are expressed in chick by non-overlapping subsets of interneurons and RGCs that form synapses in distinct IPL sublaminae. Moreover, each protein is concentrated within the appropriate sublaminae and each mediates homophilic adhesion. Loss- and gain-of-function studies in vivo indicate that these IgSF members participate in determining the IPL sublaminae in which synaptic partners arborize and connect. Thus, vertebrate Dscams, like Drosophila Dscams, play roles in neural connectivity. Together, our results on Dscams and Sidekicks suggest the existence of an IgSF code for laminar specificity in retina and, by implication, in other parts of the central nervous system.     

视网膜神经节细胞非经典感受野及其方位倾向性的模型研究

建立了包含无长突细胞相互抑制网络的视网膜神经节细胞的三层网络模型,以验证李朝义等提出的非经典感受野可能源于无长突细胞相互抑制的假说,并探讨神经节细胞感受野的方位倾向性的可能机制．模拟结果表明,通过无长突细胞之间的相互抑制可以形成神经节细胞的非经典感受野．模型模拟了神经节细胞的中心区和大周边区的方位倾向性及其相互作用,结果提示神经节细胞的方位倾向性可能主要源于神经节细胞树突野的空间分布．     

Neurite arborization and mosaic spacing in the mouse retina require DSCAM

To establish functional circuitry, retinal neurons occupy spatial domains by arborizing their processes, which requires the self-avoidance of neurites from an individual cell, and by spacing their cell bodies, which requires positioning the soma and establishing a zone within which other cells of the same type are excluded. The mosaic patterns of distinct cell types form independently and overlap. The cues that direct these processes in the vertebrate retina are not known. Here we show that some types of retinal amacrine cells from mice with a spontaneous mutation in Down syndrome cell adhesion molecule (Dscam), a gene encoding an immunoglobulin-superfamily member adhesion molecule, have defects in the arborization of processes and in the spacing of cell bodies. In the mutant retina, cells that would normally express Dscam have hyperfasciculated processes, preventing them from creating an orderly arbor. Also, their cell bodies are randomly distributed or pulled into clumps rather than being regularly spaced mosaics. Our results indicate that mouse DSCAM mediates isoneuronal self-avoidance for arborization and heteroneuronal self-avoidance within specific cell types to prevent fasciculation and to preserve mosaic spacing. These functions are analogous to those of Drosophila DSCAM (ref. 6) and DSCAM2 (ref. 7). DSCAM may function similarly in other regions of the mammalian nervous system, and this role may extend to other members of the mammalian Dscam gene family.     

Retinal pigmented epithelial cells induced to transdifferentiate to neurons by laminin

Although the regeneration of nervous tissue in the vertebrate is very limited, there are a few remarkable examples of this process. Understanding the factors that regulate CNS regeneration in those areas of the nervous system where it occurs, will doubtless provide generally applicable, essential information about the process. It has been known for some time that the amphibian retina regenerates following its destruction. Transplant studies, confirmed later by in vitro experiments, have shown that one source of new neurons in regenerating retina is the retinal pigmented epithelium (RPE). RPE cells can transdifferentiate to either neurons or lens cells in culture, but little is known about the factors that regulate this process. A recent study in vivo of retinal regeneration provided evidence that the association of RPE cells with the retinal vascular membrane is an important step in transdifferentiation. We report here that transdifferentiation in vitro is profoundly influenced by the substrate on which the cells are cultured; RPE cells plated on laminin-containing substrates frequently transdifferentiate into neurons. In addition, we have found a high concentration of laminin in the Rana retinal vascular membrane. Therefore, we propose that retinal regeneration is initiated by changes in the composition of the extracellular matrix that RPE cells contact early in the process.     

Novel pathway connecting the outer and inner vertebrate retina

In many fish retinas, thin axons from the external horizontal cells extend through the inner nuclear layer and expand into large terminal processes that lie along the border of the inner nuclear and inner plexiform layers. Although the horizontal-cell axon terminals are structurally very prominent, their function is unknown. Here we report morphological and functional evidence that signals from catfish (Ictalurus punctatus) horizontal-cell axon terminals can be transmitted directly to amacrine cells. Current injected into horizontal-cell axon terminals produces responses from both transient and sustained amacrine cells very similar to those elicited by light stimuli. Electron microscope observations show chemical synapses from the axon terminals onto amacrine cell perikarya and processes. These data suggest that amacrine cells in the catfish retina receive two inputs, one from bipolar cells and the other from horizontal-cell axon terminals.     

Myelin-associated glycoprotein in human retina

The human retina is unmyelinated, but structural similarities have been noted between Müller cells, the main glial cell type of retina, and oligodendrocytes, the myelin-forming cells of the central nervous system. We now show that antibodies against myelin-associated glycoprotein, a minor component of central and peripheral myelin so far found only in myelin and myelin-forming cells, also stain Müller cells. Immunoblot analysis of retinal proteins indicates that the antigen detected is myelin associated glycoprotein. These results suggest a closer relationship between Müller cells and oligodendrocytes than previously suspected and raise questions about the functional role of myelin-associated glycoprotein.     

Substance P-immunoreactive retinal ganglion cells and their central axon terminals in the rabbit

Retinal ganglion cells are the projection neurons that link the retina to the brain. Peptide immunoreactive cells in the ganglion cell layer (GCL) of the mammalian retina have been noted but their identity has not been determined. We now report that, in the rabbit, 25-35% of all retinal ganglion cells contain substance P-like (SP) immunoreactivity. They were identified by either retrograde transport of fluorescent tracers injected into the superior colliculus, or by retrograde degeneration after optic nerve section. SP immunoreactive cells are present in all parts of the retina and have medium to large cell bodies with dendrites that ramify extensively in the proximal inner plexiform layer. Their axons terminate in the dorsal lateral geniculate nucleus, superior colliculus and accessory optic nuclei, and these terminals disappear completely after contralateral optic nerve section and/or eye enucleation. In the dorsal lateral geniculate nucleus large, beaded, immunoreactive axons and varicosities make up a narrow plexus just below the optic tract, where they define a new geniculate lamina. The varicosities make multiple synaptic contacts with dendrites of dorsal lateral geniculate nucleus projection neurons and presumptive interneurons in complex glomerular neuropil. This is direct evidence that some mammalian retinal ganglion cells contain substance P-like peptides and strongly suggests that, in the rabbit, substance P (or related tachykinins) may be a transmitter or modulator in a specific population or populations of retinal ganglion cells.