Failure of axonal transport induces a spatially coincident increase in astrocyte BDNF prior to synapse loss in a central target.

Failure of anterograde transport to distal targets in the brain is a common feature of neurodegenerative diseases. We have demonstrated in rodent models of glaucoma, the most common optic neuropathy, early loss of anterograde transport along the retinal ganglion cell (RGC) projection to the superior colliculus (SC) is retinotopic and followed by a period of persistence of RGC axon terminals and synapses through unknown molecular pathways. Here we use the DBA/2J mouse model of hereditary glaucoma and an acute rat model to demonstrate that retinotopically focal transport deficits in the SC are accompanied by a spatially coincident increase in brain-derived neurotrophic factor (BDNF), especially in hypertrophic astrocytes. These neurochemical changes occur prior to loss of RGC synapses in the DBA/2J SC. In contrast to BDNF protein, levels of Bdnf mRNA decreased with transport failure, even as mRNA encoding synaptic structures remained unchanged. In situ hybridization signal for Bdnf mRNA was the strongest in SC neurons, and labeling for the immature precursor pro-BDNF was very limited. Subcellular fractionation of SC indicated that membrane-bound BDNF decreased with age in the DBA/2J, while BDNF released from vesicles remained high. These results suggest that in response to diminished axonal function, activated astrocytes in the brain may sequester mature BDNF released from target neurons to counter stressors that otherwise would challenge survival of projection synapses.

Neurodegeneration in glaucoma: progression and calcium-dependent intracellular mechanisms.

Glaucoma is an age-related optic neuropathy involving sensitivity to ocular pressure. The disease is now seen increasingly as one of the central nervous system, as powerful new approaches highlight an increasing number of similarities with other age-related neurodegenerations such as Alzheimer's and Parkinson's. While the etiologies of these diseases are diverse, they involve many important common elements including compartmentalized programs of degeneration targeting axons, dendrites and finally cell bodies. Most age-related degenerations display early functional deficits that precede actual loss of neuronal substrate. These are linked to several specific neurochemical cascades that can be linked back to dysregulation of Ca(2+)-dependent processes. We are now in the midst of identifying similar cascades in glaucoma. Here we review recent evidence on the pathological progression of neurodegeneration in glaucoma and some of the Ca(2+)-dependent mechanisms that could underlie these changes. These mechanisms present clear implications for efforts to develop interventions targeting neuronal loss directly and make glaucoma an attractive model for both interrogating and informing other neurodegenerative diseases.

Localization of kainate receptors to the presynaptic active zone of the rod photoreceptor in primate retina.

Visual information is encoded at the photoreceptor synapse by modulation of the tonic release of glutamate from one or more electron-dense ribbons. This release is highest in the dark, when photoreceptors are depolarized, and decreases in grades when photoreceptors hyperpolarize with increasing light. Functional diversity between neurons postsynaptic at the synaptic ribbon arises in part from differential expression of both metabotropic (G-protein-gated) and ionotropic (ligand-gated) glutamate receptor. In the brain, different subunits also modulate the presynaptic active zone. In hippocampus, ionotropic kainate receptors localize to the presynaptic membrane of glutamatergic axon terminals and facilitate depolarization of the synapse (e.g. Lauri et al., 2001). Such facilitation may be helpful in the retina, where consistent depolarization of the photoreceptor axon terminal is necessary to maintain glutamate release in the dark. We investigated whether such a mechanism could be present in primate retina by using electron microscopy to examine the localization of the kainate subunits GluR6/7 at the rod axon terminal, where only a single ribbon synapse mediates glutamate release. We scored 54 rod axon terminals whose postsynaptic space contained one or more GluR6/7-labeled processes and traced these processes through serial sections to determine their identity. Of 68 labeled processes, 63% originated from narrow "fingers" of cytoplasm extending from the presynaptic axon terminal into the postsynaptic cleft. Each rod terminal typically inserts 4-6 presynaptic fingers, and we scored several instances where multiple fingers contained label. Such consistency suggests that each presynaptic finger expresses GluR6/7. The physiological properties of kainate receptors and the geometry of the rod axon terminal suggest that presynaptic GluR6/7 could provide a steady inward current to maintain consistent depolarization of the rod synapse in the long intervals between photons in the dark.

Seeing with S cones.

The S cone is highly conserved across mammalian species, sampling the retinal image with less spatial frequency than other cone photoreceptors. In human and monkey retina, the S cone represents typically 5-10% of the cone mosaic and distributes in a quasi-regular fashion over most of the retina. In the fovea, the S cone mosaic recedes from a central "S-free" zone whose size depends on the optics of the eye for a particular primate species: the smaller the eye, the less extreme the blurring of short wavelengths, and the smaller the zone. In the human retina, the density of the S mosaic predicts well the spatial acuity for S-isolating targets across the retina. This acuity is likely supported by a bistratified retinal ganglion cell whose spatial density is about that of the S cone. The dendrites of this cell collect a depolarizing signal from S cones that opposes a summed signal from M and L cones. The source of this depolarizing signal is a specialized circuit that begins with expression of the L-AP4 or mGluR6 glutamate receptor at the S cone-->bipolar cell synapse. The pre-synaptic circuitry of this bistratified ganglion cell is consistent with its S-ON/(M+L)-OFF physiological receptive field and with a role for the ganglion cell in blue/yellow color discrimination. The S cone also provides synapses to other types of retinal circuit that may underlie a contribution to the cortical areas involved with motion discrimination.

Representation of cone signals in the primate retina.

Vision begins with specialized retinal circuits that encode diverse types of information. For Old World primates, these circuits sample three submosaics formed by cone photoreceptors sensitive to short, middle, and long wavelengths. For spatial acuity, the photon catch between any two cones is compared for discrimination of patterns as fine as the cone mosaic. For color vision, the photon catch between different cone types is compared for discrimination of fine spectral differences on the basis of hue. The retinal circuits for these two tasks differ at the synaptic level to form distinct representations of signals from the cone mosaic.

Neuronal chemistry and functional organization in the primate visual system.

Beginning with the first step of visual processing and proceeding outward from that point, the neurons involved in different aspects of vision are distinct. Stated simply, neurons doing different things look different. They often display distinct morphological features and they usually express different molecules. In addition, neurons that perform a common function usually aggregate together to form recognizable layers or compartments that can be studied in isolation because they are neurochemically distinct. Here is found, then, a junction of two major domains in neuroscience research, as discovery of molecular diversity among neurons is exploited to study organization and function of the primate visual system.

Microcircuitry and mosaic of a blue-yellow ganglion cell in the primate retina.

Perception of hue is opponent, involving the antagonistic comparison of signals from different cone types. For blue versus yellow opponency, the antagonism is first evident at a ganglion cell with firing that increases to stimulation of short wavelength-sensitive (S) cones and decreases to stimulation of middle wavelength-sensitive (M) and long wavelength-sensitive (L) cones. This ganglion cell, termed blue-yellow (B-Y), has a distinctive morphology with dendrites in both ON and OFF strata of the inner plexiform layer (Dacey and Lee, 1994). Here we report the synaptic circuitry of the cell and its spatial density. Reconstructing neurons in macaque fovea from electron micrographs of serial sections, we identified six ganglion cells that branch in both strata and have similar circuitry. In the ON stratum each cell collects approximately 33 synapses from bipolar cells traced back exclusively to invaginating contacts from S cones, and in the OFF stratum each cell collects approximately 14 synapses from bipolar cells (types DB2 and DB3) traced to basal synapses from approximately 20 M and L cones. This circuitry predicts that spatially coincident blue-yellow opponency arises at the level of the cone output via expression of different glutamate receptors. S cone stimuli suppress glutamate release onto metabotropic receptors of the S cone bipolar cell dendrite, thereby opening cation channels, whereas M and L cone stimuli suppress glutamate release onto ionotropic glutamate receptors of DB2 and DB3 cell dendrites, thereby closing cation channels. Although the B-Y cell is relatively rare (3% of foveal ganglion cells), its spatial density equals that of the S cone; thus it could support psychophysical discrimination of a blue-yellow grating down to the spatial cutoff of the S cone mosaic.

Foveal cones form basal as well as invaginating junctions with diffuse ON bipolar cells.

The response of a mammalian bipolar cell is generally thought to be determined by the location and morphology of synapses from the cone terminal: ON bipolar cells are believed to be depolarized strictly at invaginating contacts and OFF bipolar cells hyperpolarized at basal contacts. This hypothesis was re-investigated in the macaque fovea (1 deg nasal) using electron micrographs of serial sections. We determined the number of invaginating sites available and then identified the contacts to bipolar cells with axons in the ON level of the inner plexiform layer. A cone terminal forms about 20 active zones marked by ribbons. A few active zones house two invaginating dendrites, so there are 22 invaginating sites per cone. A midget ON bipolar cell collects 18 invaginating contacts from one cone, thus only about four invaginating sites remain for diffuse ON bipolar cells. Two diffuse ON cells were reconstructed; each collects about 25 contacts from an estimated 10 cones. Only three or four of these contacts are invaginating; the rest are basal, adjacent to the triad. This suggests that basal contacts can be depolarizing. The distance from the vesicle release site at active zones to an invaginating contact is 140 +/- 40 nm; to a basal contact adjacent to the triad is 500 +/- 160 nm, and to the next nearest basal contact is 950 +/- 370 nm.

Absence of spectrally specific lateral inputs to midget ganglion cells in primate retina.

Visual information is conveyed to the brain by the retinal ganglion cells. Midget ganglion cells serve fine spatial vision by summing excitation from a receptive field 'centre', receiving input from a single cone in the central retina, with lateral inhibition from a receptive field 'surround', receiving input from many surrounding cones. Midget ganglion cells are also thought to serve colour opponent vision because the centre excitation is from a cone of one spectral type, while the surround inhibition is from cones of the other type. The two major cone types, middle(M)- and long-(L)wavelength sensitive, are equally numerous and randomly distributed in the primate central retina, so a spectrally homogeneous surround requires that the cells mediating lateral interactions (horizontal or amacrine cells) receive selective input from only one cone type. Horizontal cells cannot do this because they receive input indiscriminately from M and L cones. Here we report that the amacrine cells connected to midget ganglion cells are similarly indiscriminate. The absence of spectral specificity in the inhibitory wiring raises doubt about the involvement of midget ganglion cells in colour vision and suggest that colour opponency may instead be conveyed by a different type of ganglion cell.

M and L cones in macaque fovea connect to midget ganglion cells by different numbers of excitatory synapses.

Visual acuity depends on the fine-grained neural image set by the foveal cone mosaic. To preserve this spatial detail, cones transmit through non-divergent pathways: cone-->midget bipolar cell-->midget ganglion cell. Adequate gain must be established along each pathway; crosstalk and sources of variation between pathways must be minimized. These requirements raise fundamental questions regarding the synaptic connections: (1) how many synapses from bipolar to ganglion cell transmit a cone signal and with what degree of crosstalk between adjacent pathways; (2) how accurately these connections are reproduced across the mosaic; and (3) whether the midget circuits for middle (M) and long (L) wavelength sensitive cones are the same. We report here that the midget ganglion cell collects without crosstalk either 28 +/- 4 or 47 +/- 3 midget bipolar synapses. Two cone types are defined by this difference; being about equal in number and distributing randomly in small clusters of like type, they are probably M and L.

Monochromatism determined at a long-wavelength/middle-wavelength cone-antagonistic locus.

The foveal increment threshold spectral sensitivity function for a 500 msec raised cosine stimulus without spatial edges exhibits a sharp drop or "notch" in sensitivity that coincides with the wavelength of a long-wavelength adapting field. An appropriate name for this phenomenon is the "Sloan notch", after Louise Sloan, who first observed a notch in a foveal threshold spectrum. We have examined suprathreshold discriminability on both sides of the Sloan notch produced by a 6700 td, 578 nm adapting field. In a temporal two-alternative forced-choice paradigm, a suprathreshold 650 nm low-frequency "standard" stimulus was paired with low-frequency "test" stimuli, of wavelength between 600 and 670 nm and varied intensity; the observer's task was to identify the interval containing the standard. Discriminability of the test and standard typically dropped to chance for some particular test intensity, producing "indiscriminability action spectra", up to 0.7 log units above threshold. Truncated spectra (between about 530 and 560 nm) were also obtained from observers on the middle wavelength side of the Sloan notch, for a 550 nm standard. The indiscriminability action spectra of each observer were identical, up to scaling, with the observer's threshold action spectrum. Analysis of the action spectra shows that the indiscriminable stimuli are rendered equivalent at the input to a neural pathway where L- and M-cone signals converge with opposite sign. We also investigated discriminability in the spectral region containing and immediately surrounding the Sloan notch. Suprathreshold stimuli in the spectral region near the notch produce percepts that are always discriminable from 650 and 550 nm standards (and from one another), and thus we conclude that in this spectral region, perception is mediated in part by a pathway distinct from that which signals the standards. The action spectrum of this latter pathway was estimated with a variant of the discrimination procedure, and found similar to V lambda over the spectral region 575-610 nm.