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Biomedical subjects

D I Vaney

Publications and source records attributed to D I Vaney.

At least 19 recordsLinked to original sources
PubMed2000

Dendritic computation of direction selectivity by retinal ganglion cells.

Direction-selective ganglion cells (DSGCs) in the retina respond strongly when stimulated by image motion in a preferred direction but are only weakly excited by image motion in the opposite null direction. Such coding represents an early manifestation of complex information processing in the visual system, but the cellular locus and the synaptic mechanisms have yet to be elucidated. We recorded the synaptic activity of DSGCs using strategies to observe the asymmetric inhibitory inputs that underlie the generation of direction selectivity. The critical nonlinear interactions between the excitatory and inhibitory inputs took place postsynaptically within the dendrites of the DSGCs.

Action Potentials↗
PubMed2000

The dendritic architecture of the cholinergic plexus in the rabbit retina: selective labeling by glycine accumulation in the presence of sarcosine.

The cholinergic amacrine cells in the rabbit retina slowly accumulate glycine to very high levels when the tissue is incubated with excess sarcosine (methylglycine), even though these cells do not normally contain elevated levels of glycine and do not express high-affinity glycine transporters. Because the sarcosine also depletes the endogenous glycine in the glycine-containing amacrine cells and bipolar cells, the cholinergic amacrine cells can be selectively labeled by glycine immunocytochemistry under these conditions. Incubation experiments indicated that the effect of sarcosine on the cholinergic amacrine cells is indirect: sarcosine raises the extracellular concentration of glycine by blocking its re-uptake by the glycinergic amacrine cells, and the excess glycine is probably taken-up by an unidentified low-affinity transporter on the cholinergic amacrine cells. Neurobiotin injection of the On-Off direction-selective (DS) ganglion cells in sarcosine-incubated rabbit retina was combined with glycine immunocytochemistry to examine the dendritic relationships between the DS ganglion cells and the cholinergic amacrine cells. These double-labeled preparations showed that the dendrites of the DS ganglion cells closely follow the fasciculated dendrites of the cholinergic amacrine cells. Each ganglion cell dendrite located within the cholinergic strata is associated with a cholinergic fascicle and, conversely, there are few cholinergic fascicles that do not contain at least one dendrite from an On-Off DS cell. It is not known how the dendritic co-fasciculation develops, but the cholinergic dendritic plexus may provide the initial scaffold, because the dendrites of the On-Off DS cells commonly run along the outside of the cholinergic fascicles.

Animals↗
PubMed2000

Endogenous dopaminergic regulation of horizontal cell coupling in the mammalian retina.

Horizontal cells in an isolated wholemount preparation of the mouse retina were injected with Lucifer yellow and neurobiotin to characterize both the pattern of gap junctional connectivity and its regulation by dopamine. The injected horizontal cells had a uniform morphology of a round cell body, a compact dendritic tree, and an axon, which could sometimes be traced to an expansive terminal system. The dendro-dendritic gap junctions between neighboring cells mediated both weak Lucifer yellow dye coupling and strong neurobiotin tracer coupling. The extent of the tracer coupling was decreased by either exogenous dopamine (100 microM) or cyclic adenosine monophosphate (cAMP) analogs and was significantly increased by the D1 antagonist SCH 23390 (10 microM). These results provide the first evidence in the mammalian retina that the gap junctions between horizontal cells are endogenously regulated by dopamine, which acts through D1 receptors to increase the intracellular cAMP. It has been proposed that the gap junctional coupling between horizontal cells is mediated by connexin 32 (Cx32), but the pattern and dopaminergic regulation of horizontal cell coupling were unaffected in Cx32-knockout mice, ruling out the possible involvement of Cx32. Every tracer-coupled horizontal cell showed calbindin immunoreactivity, and vice versa, providing strong evidence that the horizontal cells in the mouse retina comprise a single cell type. Like the axonless horizontal cells in other mammalian retinas, the axon-bearing horizontal cells in the mouse retina are coupled by gap junctions that are permeable to Lucifer yellow and dopamine sensitive, suggesting that the mouse horizontal cells have hybrid properties to compensate for the absence of axonless horizontal cells.

Animals↗
PubMed2000

Gap junctions in the eye: evidence for heteromeric, heterotypic and mixed-homotypic interactions.

Some of the best evidence that different types of gap junction proteins (connexins) interact with each other in vivo has been found in the eye. This review focuses on three diverse ocular tissues that may contain heterotypic or heteromeric gap junction channels. Each of the tissues uses gap junctions in a superlative fashion: The crystalline lens has an exceptionally high density of gap junctions; the ciliary body expresses a surprising variety of connexins; the neural retina shows remarkable specificity in the patterns of intercellular coupling.

Animals↗
PubMed2000

Modulation of coupling between retinal horizontal cells by retinoic acid and endogenous dopamine.

The regulation of electrical coupling between retinal neurons appears to be an important component of the neuronal mechanism of light adaptation, which enables the retina to operate efficiently over a broad range of light intensities. The information about the ambient light conditions has to be transmitted to the neuronal network of the retina and previous evidence has indicated that dopamine is an important neurochemical signal. In addition, recent studies suggest that another important chemical signal is retinoic acid, which is a light-correlated byproduct of the phototransduction cycle. This review summarizes the latest findings about the effects of dopamine and retinoic acid on gap junctional coupling in the retinas of mouse, rabbit and fish.

Animals↗
PubMed1999

Neuronal coupling in the central nervous system: lessons from the retina.

The retina is a model system for studying gap junctional intercellular communication in the CNS. The cellular coupling can be graphically visualized in retinal whole mounts by injecting small cationic tracers into microscopically identified neurons; the pattern of tracer coupling shown by each type of retina neuron is highly stereotyped, with many types of amacrine cells and ganglion cells showing complex patterns of both homologous and heterologous coupling. Parallel physiological studies have demonstrated that the gap junctions can be modulated dynamically by neurotransmitters and by the level of ambient illumination. Taken together, the numerous structural and functional studies on gap junctions in the retina provide powerful support for the concept that electrical synapses are complex components of neuronal circuits, having many of the attributes normally ascribed to chemical synapses.

Animals↗
PubMed1999

Retinoic acid modulates gap junctional permeability between horizontal cells of the mammalian retina.

In the retina, all-trans retinoic acid (at-RA) could function as a light signal because its production increases with the level of illumination. Given the well-established effects of retinoic acid on cell coupling in other tissues, it is possible that the changing levels of at-RA modulate the gap junctional permeability between retinal neurons. This study examines the effects of retinoic acid on horizontal cell coupling, which is known to be modulated by the ambient light level. Single horizontal cells were injected under visual control with either Neurobiotin (mouse retina) or Lucifer yellow (rabbit retina) and the extent of tracer coupling or dye coupling was used to monitor the gap junctional permeability. In the mouse retina, the injection of Neurobiotin revealed a network of approximately 150-250 tracer-coupled horizontal cells. The tracer coupling was completely abolished by incubating the retina in 150 microM at-RA for 35 min. In the rabbit retina, the injection of Lucifer yellow into A-type horizontal cells revealed networks of approximately 15-30 dye-coupled horizontal cells. Incubation in 150 microM at-RA reduced the dye coupling within 12 min and complete uncoupling was achieved after 35 min. The uncoupling effects of at-RA in the mouse and rabbit retinas were concentration- and time-dependent and they were reversible after washout. The coupling was not affected by either the 9-cis form of retinoic acid or by at-RA that had been isomerized by intensive light. The uncoupling effect of at-RA persisted following treatment with a D1 receptor antagonist and thus was dopamine-independent. This study has established that at-RA is able to modulate the gap junctional permeability between horizontal cells in the mammalian retina, where its light-dependent release has already been demonstrated.

Animals↗
PubMed1998

Neurotransmitter coupling through gap junctions in the retina.

Although all bipolar cells in the retina probably use the excitatory transmitter glutamate, approximately half of the cone bipolar cells also contain elevated levels of the inhibitory transmitter glycine. Some types of cone bipolar cells make heterologous gap junctions with rod amacrine cells, which contain elevated levels of glycine, leading to the hypothesis that the bipolar cells obtain their glycine from amacrine cells. Experimental support for this hypothesis is now provided by three independent lines of evidence. First, the glycine transporter GLYT1 is expressed by the glycine-containing amacrine cells but not by the glycine-containing bipolar cells, suggesting that only the amacrine cells are functionally glycinergic. Second, the gap-junction blocker carbenoxolone greatly reduces exogenous 3H-glycine accumulation into the bipolar cells but not the amacrine cells. Moreover, when the endogenous glycine stores in both cell classes are depleted by incubating the retina with a glycine-uptake inhibitor, carbenoxolone blocks the subsequent glycine replenishment of the bipolar cells but not the amacrine cells. Third, intracellular injection of rod amacrine cells with the gap-junction permeant tracer Neurobiotin secondarily labels a heterogenous population of cone bipolar cells, all of which show glycine immunoreactivity. Taken together, these findings indicate that the elevated glycine in cone bipolar cells is not derived by high-affinity uptake or de novo synthesis but is obtained by neurotransmitter coupling through gap junctions with glycinergic amacrine cells. Thus transmitter content may be an unreliable indicator of transmitter function for neurons that make heterologous gap junctions.

Amino Acid Transport Systems, Neutral↗
PubMed1998

The modulation of intercellular coupling in the retina.

Gap-junction coupling between retinal neurons of the same (homologous) or different (heterologous) type can be modulated dynamically by neurotransmitters and by the level of ambient illumination. The homologous coupling of both horizontal cells and rod (AII) amacrine cells is reduced by dopamine, but there appear to be cell and species differences in how the ambient illumination interacts with the dopaminergic system to modulate the coupling. Both the homologous coupling of horizontal cells and the heterologous coupling between AII amacrine cells and cone bipolar cells are reduced by nitric oxide, but it is not known if changes in illumination modulate coupling through an endogenous nitrergic mechanism. The heterologous coupling between rod and cone photoreceptors is modulated by illumination, but the endogenous mechanism has not been identified.

Animals↗
PubMed1995

The immunocytochemical detection of amino-acid neurotransmitters in paraformaldehyde-fixed tissues.

In this study, we show that specific antibodies can be raised against paraformaldehyde conjugates of amino acids, including the neurotransmitters glycine, gamma-amino-butyric acid and glutamate, and a non-neuroactive amino acid, glutamine. These antibodies against paraformaldehyde conjugates specifically detect the above amino acids in paraformaldehyde-fixed tissues. The penetration of antibodies into paraformaldehyde-fixed tissues is much superior to the penetration of antibodies into glutaraldehyde-fixed tissues; hence good labeling can be observed through the depth of the tissues. Unlike glutaraldehyde, fixation with paraformaldehyde does not give rise to high levels of tissue autofluorescence and, thus, these antibodies are very effective for immunofluorescence studies. Furthermore we suggest that the ability of these antibodies to detect amino acids in paraformaldehyde-fixed tissues will permit their use in situations where it is necessary to detect other other fixation-sensitive antigens, such as neurotransmitter receptors and transporters.

Amino Acids↗
PubMed1994

pH-gated dopaminergic modulation of horizontal cell gap junctions in mammalian retina.

Horizontal cells mediate lateral inhibition in the outer retina, and this process is dependent on electrical coupling through gap junctions, giving rise to receptive fields that are much wider than the dendritic fields. This study on rabbit retina shows that the permeability of the gap junctions between A-type horizontal cells, as assessed by Lucifer yellow dye coupling, is modulated by dopamine through a D1 receptor linked to adenylate cyclase. Both exogenously applied dopamine and endogenously released dopamine uncoupled the horizontal cells, but the effect was pH-gated whereby it occurred only at an extracellular pH 7.2 +/- 0.05. The horizontal cells also uncoupled in acidic media (pH 7.0 or below) in the absence of dopamine. Our results show that horizontal cell coupling in the mammalian retina is regulated by both dopamine and pH. Given that the pH in the outer retina varies with the metabolic activity of the photoreceptors, these results suggest that ambient light conditions could gate the activity of neurotransmitters through pH-sensitive mechanisms.

Adenylyl Cyclases↗
PubMed1994

Territorial organization of direction-selective ganglion cells in rabbit retina.

The On-Off direction-selective (DS) ganglion cells in the rabbit retina comprise four discrete subtypes that orthogonally code the direction of image motion. This study examined the dendritic relationships between different subtypes of On-Off DS cells, which were identified by their distinctive type 1 bistratified (BiS1) morphology following the intracellular injection of a biotinylated tracer or Lucifer yellow under direct microscopic control. The dendrites of BiS1 cells that had closely spaced somata, which presumably comprised subtypes of On-Off DS cells with different preferred directions, were not randomly superimposed but were fasciculated into loose bundles. By contrast, tracer coupling revealed that neighboring On-Off DS cells of one subtype were highly territorial, providing complete coverage of the retina with minimal overlap. This mirrors, on a larger scale, the territorial organization within the dendritic tree of individual DS cells, suggesting that similar interactions shape both the branching pattern and the spatial extent of these neurons. Moreover, the dendrites at the edge of the dendritic field often formed tip-to-shaft or tip-to-tip contacts with dendrites from coupled cells, thus appearing to form closed dendritic loops that may be equivalent to those found within the dendritic tree. Consequently, the dendrites of one subtype are distributed uniformly and economically across the retina. The resulting plexus forms a strikingly regular scaffold on which the presynaptic interneurons generate direction selectivity.

Animals↗
PubMed1993

Unidirectional coupling of gap junctions between neuroglia.

Gap junctions permit the passage of ions and small molecules between cells, thereby providing a basis for direct intercellular communication. In the rabbit retina, the low molecular weight dyes Lucifer yellow and biocytin passed readily from astrocytes into adjacent astrocytes, oligodendrocytes, and Müller cells. However, the dyes rarely passed from either oligodendrocytes or Müller cells into astrocytes. Unidirectional passage of dye suggests the presence of an asymmetric barrier to the movement of molecules through heterologous gap junctions and indicates the potential for a hierarchy of command between interconnected cells.

Animals↗
PubMed1993

The coupling pattern of axon-bearing horizontal cells in the mammalian retina.

In most vertebrate retinae, horizontal cells of the same functional type are homologously coupled through gap junctions. However, ultrastructural, physiological and dye-coupling studies have shown that the axon terminals of horizontal cells in mammalian retinae are not connected by gap junctions. In this study, intracellular injection of the junction-permeant tracer, Neurobiotin, combined with photochromic intensification of the diaminobenzidine (DAB) reaction product, has revealed that the B-type horizontal cells in rabbit retina show strong tracer coupling, both between the dendritic trees and between the axonal arborizations. These findings establish that the coupling pattern of axon-bearing horizontal cells in a mammalian retina in qualitatively similar to the coupling patterns of axon-bearing horizontal cells in non-mammalian retinae.

3,3'-Diaminobenzidine↗
PubMed1993

The spatial organization of tyrosine hydroxylase-immunoreactive amacrine cells in the chicken retina and the consequences of myopia.

We examined the spatial organization of the putative dopaminergic amacrine cells in the chicken retina and how this organization was affected by myopic eye enlargement. Myopia was produced by monocular lid suture for 4-7 months from hatching. Dopaminergic amacrine cells (TH-IR) were labelled by tyrosine hydroxylase immunohistochemistry. The somata of the TH-IR cells were usually located at the inner border of the inner nuclear layer; they gave rise to a dense plexus in stratum 1 (S1) of the inner plexiform layer, to a sparse plexus in stratum 3 (S3), and to short spiny dendrites at the border of strata 4 and 5 (S4/S5). The long thin processes in S1 and S3 could seldom be traced to their cell of origin, whereas the S4/S5 dendrites formed discrete fields that tiled the retina with little overlap. Lid suture resulted in retinal expansion of between 25-70%, but the total number of TH-IR amacrine cells was unaltered. Per retina, there were about 4700 TH-IR amacrine cells which showed a 3:1 density gradient from central to peripheral retina. The size of the S4/S5 dendritic fields increased proportionately in the expanded retinae, thus maintaining their coverage across the retina. The increase was achieved through scaled growth of the S4/S5 dendrites, involving both terminal and non-terminal dendrites. These findings suggest that the expansion of retinal neurons during myopia occurred through normal, albeit excessive, growth mechanisms.

Animals↗
PubMed1992

Photochromic intensification of diaminobenzidine reaction product in the presence of tetrazolium salts: applications for intracellular labelling and immunohistochemistry.

The diaminobenzidine (DAB) reaction product can be greatly intensified by incubating the reacted tissue in either nitro blue tetrazolium or tetranitro blue tetrazolium and then exposing the tissue to strong light. Epi-illumination through a microscope objective enables the photochromic intensification to be carried out under direct visual control, with optimal intensification taking only 10-30 s through a 20x objective. Alternatively, the whole preparation can be intensified in a few minutes by passing it back and forth under a fibre light guide. The method can be used to intensify cells that have been labelled either by immunoperoxidase techniques or with intracellular tracers such as horseradish peroxidase and neurobiotin.

Animals↗
PubMed1992

Dopaminergic modulation of gap junction permeability between amacrine cells in mammalian retina.

In mammalian retina, the rod bipolar cells synapse on the AII amacrine cells, which are therefore the third-order neurons in the rod-signal pathway. The AII amacrine cells are connected by gap junctions, both to each other and to fourth-order, On-center cone bipolar cells. They also receive synaptic input from the dopaminergic amacrine cells, and in this study, we investigated whether dopamine modulates the permeability of the gap junctions between AII amacrine cells in the isolated rabbit retina. The small biotinylated tracer Neurobiotin was injected into nuclear yellow-labeled AII cells under direct microscopic control. The extent of tracer coupling to neighboring AII cells, 40-60 min after Neurobiotin injection (0.5 nA for 60 sec), provided a standard measure of the permeability of the homologous gap junctions. Under control conditions, individual AII amacrine cells were coupled to 73 +/- 15 neighboring cells, and this was unaffected by changes in pH from 6.6 to 7.8. Exogenous dopamine significantly reduced the tracer coupling at concentrations as low as 10 nM (26 +/- 16 cells), with the effect increasing with dopamine concentration up to 10 microM (6 +/- 4 cells). The uncoupling effect of dopamine was both blocked by the selective D1 antagonist SCH-23390 (10 microM) and mimicked by the specific D1 agonist SKF-38393 (500 microM). Moreover, the AII amacrine cells were also uncoupled when the retina was incubated in forskolin (60 microM) and isobutylmethylxanthine (200 microM). Taken together, these results indicated that the uncoupling was mediated by a D1-like receptor that stimulates cAMP production. Although the selective D1 antagonist on its own did not increase tracer coupling, suggesting that there was little release of endogenous dopamine in the superfused photo-bleached retina, veratridine-evoked release of endogenous transmitters did uncouple the AII amacrine cells, and this effect was blocked by the specific D1 antagonist.

1-Methyl-3-isobutylxanthine↗