In the teleost retina the intercellular messenger nitric oxide can be synthesized by several cell types including cone photoreceptors and H1 horizontal cells, indicating a modulatory role within the outer plexiform layer, the first stage of the visual information processing. Therefore, the aim of this study was to elucidate the effects of nitric oxide on the physiology of cone horizontal cells in the intact retina. The nitric oxide donor sodium nitroprusside (0.5-2.5 mM) enhanced the light responsiveness of cone horizontal cells and reduced the degree of electrical coupling in the network. Furthermore, the spread of intracellularly injected Lucifer Yellow was restricted. The effects on light responsiveness and electrical coupling were qualitatively mimicked by 8-bromo-cGMP (0.5 mM) and could not be achieved by ferrocyanide (1 mM), the byproduct of nitric oxide liberation from nitroprusside. The effects of NO on the responsiveness of horizontal cells may be due to an action on green- and red-sensitive cones. Nitroprusside (0.1 mM) diminished the K(+)-stimulated release of endogenous dopamine by 50%, whereas the basal dopamine release was not affected, indicating that the effects on electrotonic horizontal cell coupling were not elicited by an NO-induced release of dopamine. With respect to the morphologic plasticity of the cone-horizontal cell synapse the inhibitor of endogenous nitric oxide synthesis L-nitroarginine (0.1 mM) had no influence on the formation or retraction of spinules. These results show that NO affects the responsiveness and coupling of the horizontal cell network in a dopamine-independent way.
Dying to be seen: A series of water‐soluble and highly fluorescent perylenetetracarboxdiimides (PDIs) have been synthesized in high yields by introducing charged groups into the bay region of PDI (see picture; C gray, H white, S yellow, O red). They can be used for staining living cells as they are nontoxic.
Ambient light conditions affect the morphology of synaptic elements within the cone pedicle and modulate the spatial properties of the horizontal cell receptive field. We describe here that the effects of retinoic acid on these properties are similar to those of light adaptation. Intraorbital injection of retinoic acid into eyes of dark-adapted carp that subsequently were kept in complete darkness results in the formation of numerous spinules at the terminal dendrites of horizontal cells, a typical feature of light-adapted retinae. The formation of these spinules during light adaptation is impaired in the presence of citral, a competitive inhibitor of the dehydrogenase responsible for the generation of retinoic acid in vivo. Intracellularly recorded responses of horizontal cells from dark-adapted eyecup preparations superfused with retinoic acid reveal typical light-adapted spatial properties. Retinoic acid thus appears to act as a lightsignaling modulator. Its activity appears not to be at the transcriptional level because its action was not blocked by actinomycin.
Multi-unit recording from neuronal networks cultured on microelectrode arrays (MEAs) is a widely used approach to achieve basic understanding of network properties, as well as the realization of cell-based biosensors. However, network formation is random under primary culture conditions, and the cellular arrangement often performs an insufficient fit to the electrode positions. This results in the successful recording of only a small fraction of cells. One possible approach to overcome this limitation is to raise the number of cells on the MEA, thereby accepting an increased complexity of the network. In this study, we followed an alternative strategy to increase the portion of neurons located at the electrodes by designing a network in confined geometries. Guided settlement and outgrowth of neurons is accomplished by taking control over the adhesive properties of the MEA surface. Using microcontact printing a triangular twodimensional pattern of the adhesion promoter poly-Dlysine was applied to the MEA offering a meshwork that at the same time provides adhesion points for cell bodies matching the electrode positions and gives frequent branching points for dendrites and axons. Low density neocortical networks cultivated under this condition displayed similar properties to random networks with respect to the cellular morphology but had a threefold higher electrode coverage. Electrical activity was dominated by periodic burst firing that could pharmacologically be modulated. Geometry of the network and electrical properties of the patterned cultures were reproducible and displayed long-term stability making the combination of surface structuring and multi-site recording a promising tool for biosensor applications.
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