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W D Eldred

Publications and source records attributed to W D Eldred.

At least 19 recordsLinked to original sources
PubMed2006

Cellular and subcellular localization of PDE10A, a striatum-enriched phosphodiesterase.

PDE10A is a recently identified phosphodiesterase that is highly expressed by the GABAergic medium spiny projection neurons of the mammalian striatum. Inhibition of PDE10A results in striatal activation and behavioral suppression, suggesting that PDE10A inhibitors represent a novel class of antipsychotic agents. In the present studies we further elucidate the localization of this enzyme in striatum of rat and cynomolgus monkey. We find by confocal microscopy that PDE10A-like immunoreactivity is excluded from each class of striatal interneuron. Thus, the enzyme is restricted to the medium spiny neurons. Subcellular fractionation indicates that PDE10A is primarily membrane bound. The protein is present in the synaptosomal fraction but is separated from the postsynaptic density upon solubilization with 0.4% Triton X-100. Immuno-electron microscopy of striatum confirms that PDE10A is most often associated with membranes in dendrites and spines. Immuno-gold particles are observed on the edge of the postsynaptic density but not within this structure. Our studies indicate that PDE10A is associated with post-synaptic membranes of the medium spiny neurons, suggesting that the specialized compartmentation of PDE10A enables the regulation of intracellular signaling from glutamatergic and dopaminergic inputs to these neurons.

Animals↗
PubMed2000

Localization of natriuretic peptides and their activation of particulate guanylate cyclase and nitric oxide synthase in the retina.

In the vertebrate retina, cyclic guanosine monophosphate (cGMP) mediates photoreceptor signal transduction and modulates ion channel and gap junction conductivity. Although most previous studies have focused on its synthesis by nitric oxide (NO)-sensitive soluble guanylate cyclase, cGMP is also synthesized by NO-insensitive particulate guanylate cyclases (pGC). Natriuretic peptides and their associated pGC-coupled receptors have been reported in retina, but few studies have localized these natriuretic peptides or pGCs to specific retinal cells or demonstrated that activation of pGCs by natriuretic peptides increases cGMP synthesis. In this study, we immunocytochemically localized atrial, brain, and C-type natriuretic peptide-like immunoreactivity (ANP-LI, BNP-LI, and CNP-LI, respectively) in turtle retina by using isoform specific antisera, and determined the ability of each natriuretic peptide isoform to increase cGMP-like immunoreactivity (cGMP-LI) in retinal cells. ANP-LI and CNP-LI were localized in sparsely distributed amacrine cells with thin, intermittently varicose processes in the inner plexiform layer. BNP-LI was localized to abundant somata in the inner nuclear and ganglion cell layers and in specific amacrine and horizontal cells. Stimulation of turtle eyecups with each of these natriuretic peptides increased cGMP-LI in multistratified amacrine cells by means of NO-independent mechanisms in the central retina, and in select amacrine and bipolar cells in the peripheral retina by a nitric oxide-dependent mechanism. These results indicate that natriuretic peptides can modulate the synthesis of cGMP in select retinal neurons by two distinct signal transduction pathways in a regionally specific manner.

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PubMed1999

Stimulation with N-methyl-D-aspartate or kainic acid increases cyclic guanosine monophosphate-like immunoreactivity in turtle retina: involvement of nitric oxide synthase.

In brain and retina, stimulation with excitatory amino acids (EAA) can generate nitric oxide (NO) and increase levels of cyclic guanosine monophosphate (cGMP). Because nitric oxide synthase (NOS) has been found in retinas of all species examined to date, an NO signal-transduction pathway is likely to be present in all retinas. We tested the hypothesis that stimulation of ionotropic glutamate receptors in turtle retina would result in increases in cGMP through an NOS/NO/cGMP pathway. Following in vitro incubations of turtle eye cups with the glutamate receptor agonists, N-methyl-D-aspartate (NMDA) or kainic acid (KA), we quantified the increases in cGMP-like immunoreactivity (cGMP-LI) by using enzyme-linked immunosorbant assay (ELISA) and localized the increased cGMP-LI by using an antibody against cGMP. Stimulation with NMDA or KA increased cGMP-LI in bipolar and amacrine cells as well as in some somata in the ganglion cell layer. Either KA or NMDA produced statistically significant increases in total retinal cGMP-LI by ELISA. To test the involvement of NO, we used the NOS inhibitors 7-nitroindazole and L-nitroarginine. Both inhibitors blocked virtually all of the KA- or NMDA-stimulated increases in cGMP-LI. These results indicate that activation of ionotropic glutamate receptors can increase cGMP in select retinal neurons. Differences between the agonist-evoked increases of retinal cGMP-LI suggest that there can be specificity in the activation of the NOS/NO/cGMP signal-transduction pathway by glutamate. This suggests that, in addition to short-term electrical changes, activation of ionotropic glutamate receptors also may produce longer term modulatory or metabolic effects involving NO/cGMP.

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PubMed1998

Localization of nNOS in photoreceptor, bipolar and horizontal cells in turtle and rat retinas.

Neuronal nitric oxide synthase (nNOS), an enzyme that synthesizes NO, has been found in the outer retina using light microscopic immunocytochemistry, but its subcellular localization is unknown. We used electron immunocytochemistry to examine nNOS-like immunoreactivity (nNOS-LI) in the outer plexiform layer of turtle and rat retinas. In turtle, nNOS-LI was present in some bipolar and horizontal cell processes at photoreceptor ribbon synapses and at basal junctions between photoreceptors. In rat, nNOS-LI was present in some rod bipolar and B-type horizontal cell axon terminals at rod ribbon synapses. These results indicate that in vertebrates, NO is produced by all of the major nerve cell types in the outer retina at specific synaptic contacts.

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PubMed1998

Localization of the origin of retinal efferents in the turtle brain and the involvement of nitric oxide synthase.

Previous studies have used selective neurochemical markers or retrograde tracers to localize the cells in the brain giving rise to efferents to the turtle retina. Because of the relative selectivity of the neurochemical markers or the lack of sensitivity of the previously employed retrograde tracers, these studies did not locate all the efferent cell bodies, or they could not describe the anatomy of the efferent cells. In the present study, cholera toxin B was used as a highly sensitive retrograde tracer to investigate the distribution, number, and morphology of the retinal efferent or centrifugal cell system in turtle brain. Previous studies of the turtle retina have indicated that nitric oxide synthase may be found in some retinal efferents. Therefore, we also did colocalization studies of the retrograde tracer with reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry to investigate nitric oxide as a possible transmitter used by efferent fibers and to localize these NADPH-diaphorase-positive efferent cell bodies in the turtle brain. We found that each eye received projections from approximately 40 efferent cell bodies that were located primarily in the contralateral midbrain. The majority of efferent cell bodies were centered in the isthmic tegmentum; other efferent cells extended more rostrally into the substantia nigra, and some efferent cells extended more caudally into the nucleus raphes superior. The double-label results showed that 30% of the cholera toxin B-like immunoreactive cells were also positive for NADPH-diaphorase. The location of these double-labeled cells around the locus coeruleus corresponded to the NADPH-diaphorase-positive efferent cells in the avian isthmo-optic field. The localization of NADPH-diaphorase in these efferents indicated that they may use nitric oxide to modulate retinal function.

Animals↗
PubMed1997

Synaptic inputs to identified color-coded amacrine and ganglion cells in the turtle retina.

Previous studies have proposed models of the specific synaptic circuitry responsible for color processing in the turtle retina. To determine the accuracy of these models of the circuits underlying color opponency in the inner retina of the turtle (Pseudemys scripta), we have studied the physiology, morphology, and synaptic connectivity of identified amacrine and ganglion cells. These cells were first characterized electrophysiologically and were then stained with horseradish peroxidase. Postembedding electron immunocytochemistry for gamma-aminobutyric acid (GABA) and glycine was used to reveal the neurochemical identity of their synaptic inputs. The red-ON/green, blue-OFF small-field ganglion cell, classified as G24, branched primarily in strata S1, S4, and S5 of the inner plexiform layer (IPL). Ganglion cell G24 showed a complex receptive field organized into a red-ON center surrounded by an inhibitory region, which, in turn, was surrounded by a second excitatory region. Only the center responses were color opponent. The red-OFF/green, blue-ON large-field, stellate amacrine cell, classified as A23b, stratified exclusively in stratum S2, near the S2/S3 border. The color-coded center was surrounded by a luminosity, red-sensitive surround. Synaptic input to G24 and A23b was dominated by amacrine cells (89% and 87%, respectively). G24 received significant input from amacrine cell profiles with GABA (13% of total) as well as glycine (11% of total) immunoreactivity, mostly in the proximal stratum S5 of the IPL (64% and 67% of the total GABA- and glycine-immunoreactive input, respectively). Bipolar cell synaptic input was also found predominantly in S4 and S5 (89%). In contrast, we found no glycine-immunoreactive input to A23b, and the density of the GABA-immunoreactive amacrine cell synaptic input revealed a central (15%) to peripheral (3%) gradient within the dendritic tree. The results of the present study support the previous models of the synaptic circuitry responsible for color-opponent signal processing in the inner retina of the turtle.

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PubMed1996

Quantitative anatomy, synaptic connectivity and physiology of amacrine cells with glucagon-like immunoreactivity in the turtle retina.

Although a wide variety of neuropeptides have been localized in vertebrate retinas, many questions remain about the function of these peptides and the amacrine cells that contain them. This is because many of these peptidergic amacrine cells have been studied using only immunocylochemical techniques. To address this limitation, the present study used a combination of quantitative anatomy, biochemistry and electrophysiology to examine amacrine cells in the turtle retina that contain the neuropeptide glucagon. In the turtle retina, there is a small population of 2500 glucagonergic amacrine cells, which probably represents < 1% of the total number of amacrine cells. Circular distribution statistics indicated that many of these tristratified amacrine cells had asymmetric dendritic arborizations that were radially oriented toward the retinal periphery. The cells were found to have similar dendritic coverage factors, to be distributed in a non-random arrangement in all regions of the retina, and to peak in density in the visual streak region. Electron microscopic studies indicated that glucagonergic amacrine cells made synaptic contacts primarily with other amacrine cells, and small numbers of bipolar cells. The synaptic inputs and outputs were balanced in the inner strata of the inner plexiform layer, and were biased toward synaptic outputs in the outer strata of the inner plexiform layer. These contacts involved small unlabelled synaptic vesicles, and not the large labelled dense core vesicles also found in these neurons. The biochemical studies indicated that glucagon could be released from the retina in a calcium dependent manner by high potassium stimulation. The electrophysiology found no color opponency, and the glucagonergic amacrine cells gave sustained hyperpolarizing responses to small stimulation spots and had antagonistic surrounds. The results of these studies suggest that there are significant regional specializations of glucagonergic amacrine cells, and that they may provide OFF-modulation in interactions between the ON-and OFF-centre visual pathways in the turtle retina.

Animals↗
PubMed1996

Use of the sensitive anterograde tracer cholera toxin fragment B reveals new details of the central retinal projections in turtles.

Previous anterograde degeneration and autoradiographic studies have yielded inconsistent results on the extent and target areas of the central retinal projections in turtles. We used the highly sensitive anterograde and retrograde tracer, cholera toxin B fragment (CTB), to re-examine the central retinal projections in Pseudemys scripta elegans. In contrast to the results of the previous anterograde degeneration studies and autoradiographic studies, immunohistochemical detection of CTB-labeled central retinal axons and terminals revealed these projections with great morphological detail and clarity. In addition, the CTB labeling revealed much more extensive projections than previously realized to all known major retinorecipient areas, particularly in the dorsal and ventral lateral geniculate nuclei, the pretectal region, the nucleus of the basal optic root and the optic tectum. Although the contralateral projections were always much more extensive, ipsilateral projections were clearly present (and in some cases abundant) in all previously known retinorecipient areas. In addition, prominent contralateral (and lesser ipsilateral) retinal projections to several novel target, including the basal hypothalamus, the perirotundal nuclei, the suprapeduncular nucleus and the region between the pretectal nuclei and the tegmentum, were observed. Our results suggest that the use of CTB as a tracer to examine central retinal projections in diverse other species will show that central retinal projections are much more widespread and extensive than previously realized.

Animals↗
PubMed1995

Neurotransmitter modulation of Fos- and Jun-like proteins in the turtle retina.

The expression of the Fos and Jun families of nuclear phosphoproteins can be induced by a variety of extracellular stimuli and is known to participate in the transcriptional regulation of target genes. To examine the role of these transcription factors in retinal function, we used polyclonal antisera to localize these protein families in the turtle retina. Fos-like immunoreactivity was in many somata in the inner nuclear and ganglion cell layers. In contrast, Jun-like immunoreactivity was in a smaller number of amacrine cells and many somata in the ganglion cell layer. The monostratified dendritic arbors of one prominent amacrine cell type with Jun-like immunoreactivity were also labeled. There were no dramatic differences in the levels of Fos-like immunoreactivity or Jun-like immunoreactivity between light- or dark-adapted retinas. We examined the effects of excitatory amino acids and gamma-aminobutyric acid (GABA) on the expression of these proteins in vitro. In some experiments, cobalt was used to block synaptic transmission. The excitatory amino acids increased both Fos- and Jun-like immunoreactivity, while GABA generally showed no such stimulatory effect. In cobalt-treated retinas, the same cell types had Jun-like immunoreactivity as seen in the controls, but overall levels of immunoreactivity were increased. In cobalt-treated dark-adapted retinas, some excitatory amino acids increased cytoplasmic Fos-like immunoreactivity in the somata and processes of large cells in the ganglion cell layer. Our results suggest that Fos- and Jun-related proteins may play an important role in the postsynaptic responses to amino acid transmitters in a wide variety of amacrine and ganglion cells.

Animals↗
PubMed1995

Determination of fractal dimension of physiologically characterized neurons in two and three dimensions.

Although there is a growing interest in the application of fractal analysis in neurobiology, questions about the methodology have restricted its wider application. In this report we discuss some of the underlying principles for fractal analysis, we propose the cumulative-mass method as a standard method and we extend the applicability of fractal analysis to both 2 and 3 dimensions. We have examined the relationship between the method of log-log Sholl analysis and fractal analysis and have found that they correlate well. Measurements of physiologically characterized retinal ganglion cells indicate that different cell types can have significantly different fractal dimensions. Such differences may allow the correlation of the physiological type of a neuron with its morphological fractal dimension.

Animals↗
PubMed1994

Complexity and scaling properties of amacrine, ganglion, horizontal, and bipolar cells in the turtle retina.

In the present study we have evaluated the complexity and scaling properties of the morphology of retinal neurons using fractal dimension as a quantitative parameter. We examined a large number of cells from Pseudemys scripta and Mauremys caspica turtles that had been labeled using Golgi-impregnation techniques, intracellular injection of Lucifer Yellow followed by photooxidation, intracellular injection of rhodamine conjugated horseradish peroxidase, or intracellular injection of Lucifer Yellow or horseradish peroxidase alone. The fractal dimensions of two-dimensional projections of the cells were calculated using a box counting method. Discriminant analysis revealed fractal dimension to be a significant classification parameter among several other parameters typically used for placing turtle retinal neurons in different cell classes. The fractal dimension of amacrine cells was significantly correlated with dendritic field diameters, while the fractal dimensions of ganglion cells did not vary with dendritic field span. There were no significant differences between the same cell types in two different turtle species, or between the same types of neurons in the same species after labeling with different techniques. The application of fractal dimension, as a quantitative measure of complexity and scaling properties and as a classification criterion of neuronal types, appears to be useful and may have wide applicability to other parts of the central nervous system.

Animals↗
PubMed1994

Anatomical characterization of retinal ganglion cells that project to the nucleus of the basal optic root in the turtle (Pseudemys scripta elegans).

There is little detailed information about retinal ganglion cells which project to specific central targets in the brain. The present study examined retinal ganglion cells projecting to the nucleus of the basal optic root, a major accessory retinal target in the turtle. These ganglion cells were first selectively labeled using retrograde transport of rhodamine injected stereotaxically into the nucleus of the basal optic root. The number and distribution of the retrogradely labeled cells in the retina was then determined. Some of these retrogradely labeled cells were then injected intracellularly with Lucifer Yellow, photoconverted using diaminobenzidine, and drawn in detail using a camera lucida attachment. There were approximately 1500 ganglion cells in each retina which projected to the nucleus of the basal optic root, of which 20% had cell bodies displaced to the inner nuclear layer. More than 50% of the total population was concentrated in the visual streak region. All ganglion cells projecting to the nucleus of the basal optic root, both normal and displaced, had monostratified dendritic arborizations in stratum 1 of the inner plexiform layer. About 41% of these ganglion cells had elongated dendritic arborizations with distinct orientations, which may suggest a correlation of morphology and function. There were similarities in the overall appearance, and in the type and stratification of the dendritic arborizations of all ganglion cells projecting to the nucleus of the basal optic root. These anatomical similarities are consistent with the previously demonstrated similarities in physiology and may reflect a common role for these ganglion cells in visual processing.

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PubMed1993

Effects of excitatory amino acids on immunocytochemically identified populations of neurons in turtle retina.

Excitatory amino acids play an important role in visual processing in the retinas of many species, but little is known about the identity of the specific postsynaptic cell types and the pharmacology of their receptors. To investigate which specific cell types were affected by excitatory amino acids, we examined the effects of exogenous aspartate, glutamate, kainic acid, N-methyl-D-aspartate, and MK-801 on retinal neurons. Specific populations of neurons were labelled using antibodies directed against glucagon, enkephalin, neurotensin, gamma-aminobutyric acid, glutamic acid decarboxylase, serotonin, glycine, glutamate or aspartate. We analyzed a combination of long-term in vivo injections (seven days following an intraocular injection of kainic acid) and short term in vitro incubations. There were changes in the labelling intensity and sometimes in the relative localization of all of the antigens in the drug treated retinas. Some observations suggested that the drugs were altering neurotransmitter metabolism. Differential responses were seen in specific cell types within the populations of neurons with neurotensin-, glutamate-, aspartate-, glycine, gamma-aminobutyric acid-, and glutamic acid decarboxylase-like immunoreactivity. The immunocytochemical approach used in these studies was able to determine specific retinal cell types which were influenced by particular excitatory amino acids. The broad extent of cell types influenced and the potential metabolic effects suggest that excitatory amino acids and their receptors play a complex role in visual processing.

Amino Acids↗
PubMed1992

Colocalization of enkephalin-, glucagon-, and corticotropin-releasing factor-like immunoreactivity in GABAergic amacrine cells in turtle retina.

The large number of amacrine cells which contain gamma-aminobutyric acid (GABA) in the turtle retina makes it difficult to examine specific GABAergic cell types. In order to selectively label subpopulations of GABAergic neurons, we have used fluorescent double-labeling immunocytochemical techniques to examine the localization of GABA-like immunoreactivity (LI) in amacrine cells which contain antigens resembling the neuropeptides glucagon (GLUC), corticotropin-releasing factor (CRF) or enkephalin (ENK). GABA-LI was found in 41% of the cells with GLUC-, 100% of the cells with CRF-, and 69% of the cells with ENK-LI. There were regional differences in the presence of GABA-LI in amacrine cell populations with ENK-LI. GABA-LI was present in about 80% of the cells with ENK-LI outside of the visual streak, while only 37% of the cells within the streak had GABA-LI. Based on the distinct morphologies and regional distributions of these peptidergic amacrine cells, we conclude that they represent different subpopulations of GABAergic amacrine cells in the turtle retina. Future studies can now utilize existing information regarding the synaptic connectivity of these peptidergic amacrine cells to help delineate the functions of GABAergic amacrine cells in the turtle retina.

Animals↗
PubMed1992

Effect of viscosity on neurite outgrowth and fractal dimension.

The growth mechanism by which neurons achieve their characteristic ramified morphology has long been of interest, but determining whether physical parameters, such as viscosity, are important has been difficult due to a lack of useful hypotheses and standard reproducible techniques. We have recently shown that neurons exhibit fractal behavior and that their fractal dimension (df) is consistent with a physical process called diffusion-limited aggregation (DLA). We suggested that this DLA behavior might stem from viscosity differences, chemical gradients or electrical fields (Caserta et al., Phys. Rev. Lett., 64 (1990) 95-98). DLA is a model for a large family of growth processes. In order for a process to fit the DLA model, the growth rate must be proportional to the gradient of a field at a point on the growing structure (Feder, Plenum, New York, 1988, Ch. 4). Chemical, electrical, or fluid pressure fields can fit the model depending on the particular physical system under study. Here, we studied growth of retinal neurons from chick embryos in culture media of various fluid viscosities. Thus, we test whether DLA in this system was based on a fluid pressure field. As viscosity was increased from 1 to 4.3 cps, the number of neurite branches decreased 98%. However, there was no effect on df. Over this range of viscosities, total cellular protein synthesis decreased only 17%. The results indicate that, while differences in viscosity between the interior and exterior of the cell affect neurite outgrowth, they do not affect the fractal behavior of neurons. Thus, viscosity differences are not the basis for the DLA pattern of neuronal arborization.

Algorithms↗
PubMed1992

Immunocytochemical and biochemical studies of histamine in the retina of the turtle Pseudemys scripta.

A combination of immunocytochemical and biochemical methods was used to study histamine in the turtle retina. Histamine-like immunoreactivity was localized within paraboloids of certain cone photoreceptors by use of two different antisera directed against histamine. Preincubation of eyecups in Ringer's containing 10 microM histamine selectively increased the immunoreactivity of these photoreceptor paraboloids. The present localization of histamine in paraboloids indicated that, although histamine is in photoreceptors of the turtle retina, it may play some metabolic or neuromodulatory role, and not function as a neurotransmitter.

Animals↗
PubMed1991

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

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

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