Biomedical subjects

Franklin R Amthor

Publications and source records attributed to Franklin R Amthor.

7 recordsLinked to original sources

PubMed2005

Rabbit retinal ganglion cells express functional alpha7 nicotinic acetylcholine receptors.

It is well known that cholinergic agents affect ganglion cell (GC) firing rates and light responses in the retinas of many species, but the specific receptor subtypes involved in mediating these effects have been only partially characterized. We sought to determine whether functional alpha(7) nicotinic acetylcholine receptors (nAChRs) contribute to the responses of specific retinal GC classes in rabbit retina. We used electrophysiology, pharmacology, immunohistochemistry, and reverse transcriptase-polymerase chain reaction to determine the pharmacological properties and expression of nAChR subtypes by specific rabbit retinal GC classes. Choline was used as an alpha(7) nAChR agonist. Methyllycaconitine (MLA) was used as a competitive alpha(7) nAChR antagonist. The application of choline before synaptic blockade resulted in changes in retinal GC activity, including increases or decreases in maintained firing and/or enhancement or suppression of light responses. Many physiologically identified GC types, including sustained off, sustained on, transient off, and transient on cells, demonstrated responses to choline application while under synaptic blockade. The choline-induced responses could be blocked with MLA, confirming alpha(7) nAChR activation. Individual choline-responsive GCs displayed mRNA transcripts consistent with the expression of functional alpha(7) nAChRs. Other GCs demonstrated physiological responses and mRNA expression consistent with the expression of both alpha(7) and non-alpha(7) nAChRs. Thus mRNA is present for multiple nAChR subunits in whole retina extracts, and functional alpha(7) nAChRs are capable of modulating the responses of GCs in adult rabbit retina. We also demonstrate through physiological responses that subsets of GCs express more than one nAChR subtype.

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PubMed2003

A new transparent multi-unit recording array system fabricated by in-house laboratory technology.

A new recording array system has been developed to record multi-unit activity in rabbit retina. The array consists of individually laid down layers of carbon fiber or tungsten microelectrodes whose center-center spacing can be made less than 100 microm. The array and associated electronics can be constructed by technology typically found in most electrophysiology laboratories. The array is mostly transparent, so that visual stimuli and microscopic examination can take place through it. The array can be manipulated much like a single electrode, and thus can be used to record from multiple tissue sites. Arrays as large as 32 elements have been used, with success rates of about 50% per electrode, with some electrodes picking up more than one cell. Stable recordings have been held for up to 6 h from groups of ganglion cells in an isolated eyecup preparation. These multi-electrode arrays have been used repeatedly in experiments for several months without any obvious degradation in recording quality. Although the arrays are hand-made, their layered method of assembly allows as many as 32 elements to be assembled.

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PubMed2003

Synaptic input to the on-off directionally selective ganglion cell in the rabbit retina.

A physiologically identified on-off directionally selective (DS) ganglion cell with its preferred-null axis defined was stained with horseradish peroxidase (HRP) and prepared for electron microscopy. A continuous series of thin sections were used to examine the cell's synaptology. Although the DS cell dendrite received the majority of its synaptic input from a heterogeneous population of amacrine cell processes, a frequently observed synaptic profile consisted of a DS cell dendrite receiving synapses from a cluster of several amacrine cell processes. These clusters of processes were assumed to be from a fascicle of amacrine cells, most of which probably belonged to several different cholinergic starburst amacrine cells. The most frequently observed presynaptic profile within the clusters consisted of a synaptic couplet in which two processes synapsed with each other before one of them finally synapsed with the DS ganglion cell dendrite; occasionally, a chain of three serial synapses was seen. In addition, a specific microcircuit that has the potential to exert lateral feedforward inhibition was also observed. This microcircuit consisted of two cone bipolar cell terminal dyad synapses where one dyad contained an amacrine cell process making a reciprocal synapse and a DS ganglion cell dendrite receiving direct excitation; the other dyad synapse, found lateral to the first dyad, contained two amacrine cell processes that both made reciprocal synapses, but one fed forward to make a putative inhibitory synapse with the DS cell dendrite.

Amacrine Cells↗

PubMedDate not supplied

Effects of the destruction of starburst-cholinergic amacrine cells by the toxin AF64A on rabbit retinal directional selectivity.

The effects of intraocular injections of ethylcholine mustard aziridinium ion (AF64A), an irreversible inhibitor of choline uptake, on the rabbit retina were assessed electrophysiologically, pharmacologically, anatomically, and behaviorally. Survival times from 1 day to 30 days were investigated. After 24 h, the shortest time tested, the directional selectivity of On-Off responding ganglion cells having the characteristic morphology of On-Off directionally selective directionally selective (DS) ganglion cells, as revealed by intracellular dye injection, was significantly reduced, both by an apparent decrease of preferred direction responses and an increase in responses to null-direction movement. No toxin-mediated changes in the dendritic trees of these cells were noted. Cells in AF64A-affected retinas having the DS morphology did not respond significantly to GABAergic or cholinergic agents such as picrotoxin and eserine, but did respond to nicotine. Recordings from a small random sample of other ganglion cell classes in the same retinas yielded no obvious changes in response properties. The direct effects on starburst (cholinergic) amacrine cells, which were identified by intraocular injection of the fluorescent dye DAPI with the AF64A, were investigated by intracellular injections of Lucifer yellow, and by immunohistochemical staining with antibodies to choline acetyltransferase (ChAT). Although starburst amacrine cell somas survived the AF64A treatment for at least several days, the dendrites could not be visualized by fluorescent dye injection in affected retinas due to dye leakage of the injected fluorescent dye from either the soma or proximal dendritic region. ChAT staining revealed a sequence in which ChAT-positive cells were undetectable first in the inner nuclear layer, and then in the ganglion cell layer. Cholinergic amacrine cells in the central retina were also affected before those in the periphery. The electrophysiological changes observed typically preceded the loss of ChAT activity. Behavioral tests for optokinetic nystagmus responses also revealed a lack of such responses in the affected eyes.

Amacrine Cells↗

PubMedDate not supplied

Rabbit retinal ganglion cell responses to nicotine can be mediated by beta2-containing nicotinic acetylcholine receptors.

Acetylcholine (ACh) affects the response properties of many retinal ganglion cells (GCs) through the activation of nicotinic acetylcholine receptors (nAChRs). To date there have been few studies directly correlating the expression of specific nAChR subtypes with the physiological and morphological characteristics of specific retinal GCs. This study was designed to correlate responses to nicotine application with immunohistochemical evidence of nAChR expression in physiologically and morphologically identified ganglion cells. Extracellular recordings were used to physiologically identify rabbit retinal GCs, based on responses to light stimulation. Cells were then tested for responses to nicotine application and/or for expression of nAChRs, as judged by immunoreactivity to mAb210, an nAChR antibody. The morphologies of many physiologically identified cells were also determined by dye injection. More than three-fourths of ganglion cells tested responded to nicotine application under cobalt-induced synaptic blockade. The nicotine sensitivity was consistent with nAChR immunoreactivity and was also correlated with specific morphological subgroups of GCs. Overall, approximately two-thirds of all physiologically identified GCs that were processed for immunohistochemistry displayed immunoreactivity. In total, 18 of 22 physiologically identified cells demonstrated both sensitivity to nicotine application under synaptic blockade and mAb210 immunoreactivity (mAb210-IR). Thus, mAb210-IR is likely to represent functional nAChRs that can modulate retinal information processing and visual functioning via direct excitation of a number of GC classes.

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PubMedDate not supplied

Stimulus-dependent correlated firing in directionally selective retinal ganglion cells.

Synchronous spiking has been postulated to be a meta-signal in visual cortex and other CNS loci that tags neuronal spike responses to a single entity. In retina, however, synchronized spikes have been postulated to arise via mechanisms that would largely preclude their carrying such a code. One such mechanism is gap junction coupling, in which synchronous spikes would be a by-product of lateral signal sharing. Synchronous spikes have also been postulated to arise from common-source inputs to retinal ganglion cells having overlapping receptive fields, and thus code for stimulus location in the overlap area. On-Off directionally selective ganglion cells of the rabbit retina exhibit a highly precise tiling pattern in which gap junction coupling occurs between some neighboring, same-preferred-direction cells. Depending on how correlated spikes arise, and for what purpose, one could postulate that synchronized spikes in this system (1) always arise in some subset of same-direction cells because of gap junctions, but never in non-same-preferred-directional cells; (2) never arise in same-directional cells because their receptive fields do not overlap, but arise only in different-directional cells whose receptive fields overlap, as a code for location in the overlap region; or (3) arise in a stimulus-dependent manner for both same- and different-preferred-direction cells for a function similar to that postulated for neurons in visual cortex. Simultaneous, extracellular recordings were obtained from neighboring On-Off directionally selective (DS) ganglion cells having the same and different preferred directions in an isolated rabbit retinal preparation. Stimulation by large flashing spots elicited responses from DS ganglion-cell pairs that typically showed little synchronous firing. Movement of extended bars, however, often produced synchronous spikes in cells having similar or orthogonal preferred directions. Surprisingly, correlated firing could occur for the opposite contrast polarity edges of moving stimuli when the leading edge of a sweeping bar excited the receptive field of one cell as its trailing edge stimulated another. Pharmacological manipulations showed that the spike synchronization is enhanced by excitatory cholinergic amacrine-cell inputs, and reduced by inhibitory GABAergic inputs, in a motion-specific manner. One possible interpretation is that this synchronous firing could be a signal to higher centers that the outputs of the two DS ganglion cells should be "bound" together as responding to a contour of a common object.

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PubMedDate not supplied

Retinal ganglion cell coding in simulated active vision.

The image on the retina is almost never static. Eye, head, and body movements, and externally generated motion create rapid and continual changes in the retinal image ("active vision"). Virtually all vision in animals such as primates, which make saccades as often as 3-4 times/s, is based on information that must be derived from the first few hundred milliseconds after sudden, global changes in the retinal image. These changes may be accompanied by large changes in area mean luminance, as well as higher order image contrast statistics. This study investigated how retinal ganglion cell responses, whose response properties have been typically studied and defined in a stable stimulus regime, are affected by sudden changes in mean luminance that are characteristic of active vision. Specifically, the steady-state responses of retinal ganglion cells to static or moving square-wave grating stimuli were recorded in an isolated, superfused rabbit eyecup preparation and compared to responses after saccade-like changes in luminance. The manner of coding after luminance changes was different for different ganglion cell classes; both suppression and enhancement of responses to patterns following luminance changes were found. Brisk-transient Off cells unambiguously signaled the darkening of the overall image, but were also modulated by the subsequently appearing grating stimulus. Several types of On-center cell behavior were observed, ranging from strong suppression of the subsequent response by luminance changes, to strong enhancement. Overall, most ganglion cells distinguished static patterns after a luminance change via differences in their spike discharges nearly as well as before, although there were clear asymmetries between the On and Off pathways. Changes in mean luminance in some ganglion cells, such as On-Off directionally selective ganglion cells, could create large phase shifts in the response to patterned, moving stimuli, although these stimuli were still detected immediately after luminance changes. The results of this study show that the image dynamics of active vision may be a fundamental challenge for the visual system because of strong effects on retinal ganglion cell function. However, rapid extraction of unambiguous information after luminance changes appears to be encoded in differences in the spike discharges in different retinal ganglion cell classes. Asymmetries among ganglion cell classes in sensitivity to luminance changes may provide a basis by which some provide the "context" for interpreting the firing of others.

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