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PubMed1998

The patterns of retinal ganglion cell death in hypertensive eyes.

We have recently described a rat model of hypertensive eye in which cauterizing limbal derived episcleral veins leads to increase in the intraocular pressure [S.R. Shareef, E. Garcia-Valenzuela, A. Salierno, J. Walsh, S.C. Sharma, Chronic ocular hypertension following episcleral venous occlusion in rats, Exp. Eye Res. 61 (1995) 379-382.]. We have further documented that retinal ganglion cell death is apoptotic [E. Garcia-Valenzuela, S. Shareef, J. Walsh, S.C. Sharma, Programmed cell death of retinal ganglion cells during experimental glaucoma, Exp. Eye Res. 61 (1995) 33-44.]. Here, we describe the total loss of retinal ganglion cells at various time intervals following increased IOP. At early time points death of ganglion cells in the central, peripheral retina occurred with similar frequencies. Between 4-6 weeks after intraocular elevation, ganglion cells in the peripheral retina were more susceptible than the central retina. Percentage of total ganglion cell death over the 10 week period was presumably linear and was about 4% per week.

Animals↗
PubMed1998

Peripherin-like immunoreactivity in type II spiral ganglion cell body and projections.

Peripherin, an intermediate filament protein, is present in neuronal subpopulations of both peripheral and central nervous systems. The distribution of peripherin was studied in the adult rat cochlea using immunohistochemistry on whole mount material, in cryostat sections and sections of plastic embedded tissue. In the spiral ganglion, peripherin labeling was restricted to the perikarya of a subpopulation of neurons and their peripheral and central processes. Peripherin positive neurons had the following features: (i) they have a large eccentric nucleus, they were often found in a cluster of 2 or 3 cells, (ii) they were often located near the intraganglionic spiral bundle fibers, (iii) they represented roughly 8% of the whole ganglion population and (iv) on the average they had smaller perikarya than non-immunoreactive cells. Immunostaining on semithin plastic sections revealed positive reactivity on Type II ganglion cells, while Type I neurons were negative. Double labeling using peripherin and three neurofilament (NF) subunit antibodies confirmed the presence of both markers within the same spiral ganglion cell type. Type II neurons have been previously documented as the only subpopulation of the spiral ganglion that presents a strong positive NF immunoreactivity within their perikarya. In the organ of Corti, peripherin-positive fibers formed bundles that course beneath the outer hair cells and send branches that end as boutons contacting the outer hair cells. All these characteristics suggest that peripherin-positive cells are Type II neurons, and that peripherin constitutes a reliable marker for this spiral ganglion subpopulation, as well as their peripheral and central processes.

Animals↗
PubMed2001

Glutamate-mediated responses in developing retinal ganglion cells.

Glutamate has been suggested to regulate the development of retinal ganglion cells, but little is known about the functional properties of glutamate receptors during ontogeny of these neurons. Using whole-cell and outside-out patch-clamp recordings, we have characterized the pharmacological, rectification, and kinetic properties of ionotropic glutamate receptors in ganglion cells isolated from fetal and postnatal cat retinas. The fetal cells were studied at embryonic day 33 (E33) to E38, before significant outgrowth of their dendritic processes and prior to the formation of synaptic contacts in the inner plexiform layer. In several respects, the functional properties of early fetal ganglion cells were found to be remarkably similar to those of postnatal cells. In both age groups, glutamate and AMPA produced fast desensitizing currents, kainate yielded large steady-state currents, while applications of NMDA resulted in multiple channel openings. The shapes and amplitudes of these glutamate-gated currents were also similar and the current-voltage relations were nearly linear, with reversal potentials near 0 mV. Moreover, the dose-response curves (to kainate) were virtually identical in the fetal and postnatal neurons. The proportion of neurons responsive to NMDA and non-NMDA agonists was nearly the same in both age groups. This early functional expression of glutamate receptors cannot be involved in the transmission of electrical information in the developing retina because at this stage few ganglion cells are capable of generating action potentials (Skaliora et al., 1993). It is suggested that the early activation of NMDA and non-NMDA receptors in fetal ganglion cells may regulate the outgrowth and stabilization of dendritic processes in these neurons. Our data also revealed some differences in the responses of fetal and postnatal cells to glutamate and its agonists. Thus, the unitary NMDA conductance was found to decrease with age, while the rate of glutamate receptor desensitization increased with age. Also, while virtually all postnatal cells responded to glutamate, the proportion of fetal cells that manifested glutamate-mediated responses was lower. These maturational changes presumably allow retinal ganglion cells to integrate synaptic inputs for the transmission of electrical signals to the visual centers of the brain.

Animals↗
PubMed2001

Transient expression of somatostatin mRNA in developing ganglion cell layers of rat retina.

Somatostatin (SOM) mRNA in developing ganglion cell layer (GCL) detected by in situ hybridization histochemistry and SOM peptide in developing optic chiasma and optic tract detected by immunocytochemistry were monitored to explore whether ganglion cells expressing SOM project to the visual center. Most of these cells in the developing GCL expressed SOM transiently from embryonic day 13 (E13) to E21. The cells expressing SOM mRNA initially followed a central-to-peripheral pattern of development. The cells expressing SOM mRNA in the retinas of fetuses became detectable at E13. From E14 to E17 the number of cells expressing SOM mRNA increased rapidly. At E17 most of the cells in the developing GCL expressed SOM mRNA. From E18 to postnatal days the positive cells became sparse except at the postnatal day 0 (PND0) the positive cells decreased dramatically in comparison with that at the E21. At PND15, the positive cells only can be found in the inner neuroblastic layer and in the ganglion cell layer. At PND20 the distribution pattern and the number of the positive cells were essentially the same as that in adult rat. SOM immunoreactivity was detectable at E16 in the developing optic chiasma and optic tract; the majority of the fibers in these area were SOM positive. From E16 to E18 the density of the immunostaining increased rapidly, whereas from E19 to E21 the density decreased. At PND0 no positive fibers were seen. The transient presence of SOM in most of the ganglion cells in the developing ganglion cell layer has prompted us to study the role of SOM in generation and differentiation of the retinal ganglion cells, and formation of the retina-visual center projections.

Aging↗
PubMed1999

Developmental changes in voltage-activated potassium currents of rat retinal ganglion cells.

Ca2(+)-independent voltage-activated potassium currents were investigated during the differentiation of rat retinal ganglion cells. Whole-cell patch-clamp recordings of Ca2(+)-independent voltage-activated potassium currents and their individual current components, i.e. a sustained, tetraethylammonium-sensitive current, a transient, 4-aminopyridine-sensitive current, and a slowly decaying current that was blocked by Ba2+, revealed distinct ontogenetic modifications in current densities and in activation and inactivation parameters. All three current types were expressed simultaneously at embryonic day 17/18 and were present in all retinal ganglion cells thereafter without showing any significant changes until the end of the first postnatal week. Ca2(+)-independent voltage-activated potassium current densities then increased strongly from postnatal day 8 onwards. Tetraethylammonium-sensitive current density increased about eightfold from 74 pA/pF in embryonic stages to 586 pA/pF in adult cells, whereas the transient potassium currents blocked by 4-aminopyridine increased only about 2.5-fold from 174 pA/pF to 442 pA/pF. The Ba2(+)-sensitive current increased simultaneously from 35 pA/pF to 332 pA/pF. The much higher increase in the sustained current components during retinal ganglion cell differentiation accounted for the changes in decay kinetics of Ca2(+)-independent voltage-activated potassium current observed in later postnatal stages. Alterations in current densities were paralleled by pronounced changes in current kinetics. From postnatal day 8 onwards, activation of Ca2(+)-independent voltage-activated potassium current was right-shifted for about 10 mV owing to a shift in tetraethylammonium-sensitive current-activation, whereas activation of other K+ components remained unaltered. Tetraethylammonium-sensitive current steady-state inactivation was incomplete at all developmental stages. About 50% of the tetraethylammonium-sensitive current elicited by a depolarization to +36 mV did not inactivate after prepulse potentials positive to -10 mV. In contrast, transient potassium current blocked by 4-aminopyridine almost fully inactivated during embryonic stages, whereas in adult retinal ganglion cells about 40% of this current component did not inactivate after prepulse potentials positive to -20 mV. Parallel investigation of the resting membrane potential during retinal ganglion cells differentiation showed an exponential increase from -3 mV at embryonic day 15/16 when no voltage-activated ion currents were expressed to a final value of -58 mV at postnatal day 8. These results show that fundamental potassium current modifications occur relatively late in retinal ganglion cell development and only after the resting potential is at its final value.

4-Aminopyridine↗
PubMed1997

Quantitative and topographical study of retinal ganglion cells in the chameleon (Chameleo chameleon).

Chameleons display a number of well-described physiological peculiarities of their visual system, but there is no information on the topography of the retinal ganglion cell layer. In the present study, ganglion cell density of the chameleon retina was constructed from whole mounts of the retina stained with cresyl violet. For the identification of ganglion cells, these latter cells were labelled retrogradely with horseradish peroxidase applied to the optic nerve. Using this criterion, the proportion of ganglion cells was estimated to represent 80% of retinal cells, while glial cells and amacrine cells represented 14 and 6%, respectively, of the total cell population of the retina. As for the main features of the retinal map, first, ganglion cells were distributed inhomogeneously within the ganglion cell layer, and revealed the existence of a putative area centralis. Second, a horizontal visual streak, which showed two peak density areas, was identified. These features point out the degree of specialisation of the chameleon retina and the complexity of its visual system.

Animals↗
PubMed1997

Quantitative evaluation of cochlear neurons and computer-aided three-dimensional reconstruction of spiral ganglion cells in humans with a peripheral loss of nerve fibres.

Quantitative data on human cochlear neuronal elements were collected from various regions in five patients with high-tone hearing loss due to presbycusis and in two patients with normal hearing. The number of nerve fibers was assessed in the spiral lamina and in the inner acoustic meatus together with counts of spiral ganglion cells. The results show that the number of neurons decreased peripherally, i.e., with increasing distance from the central nervous system in patients with high-tone hearing loss due to presbycusis. In two patients with normal hearing no significant difference in the number of neurons was found in the lamina spiralis as compared to the inner acoustic canal. Computer-aided 3-dimensional reconstruction of the human spiral ganglion displayed large bipolar neurons (type I cells), but also large ganglion cells with one missing axon. The results may indicate that a slow retrograde degeneration occurs from the periphery towards the spiral ganglion in presbycusis. Transmission electron microscopy analysis of freshly fixed human spiral ganglions displayed interneural connections. It is speculated whether a trophic supply from other neurons at the level of the spiral ganglion can prevent or delay further degeneration of the central axon.

Aging↗
PubMed2002

Single cell RT-PCR analysis of tyrosine kinase receptor expression in adult rat retinal ganglion cells isolated by retinal sandwiching.

We describe a protocol for analysis of gene expression in single, acutely dissociated adult rat retinal ganglion cells using RT-PCR. Retrograde tracing of retinal ganglion cells from the superior colliculi was conducted using Fluorogold. Retinas were dissected and ganglion cells isolated using retinal layer separation (sandwiching). Single, fluorescently labelled retinal ganglion cells were aspirated using a micropipette and used for PCR. Two PCR protocols are described where single cell cDNA was analysed for TrkB and GAPDH or TrkB, TrkC, Ret, Met, ErbB2 and Beta-actin by multiplex-PCR. All five tyrosine kinase receptors were amplified from single retinal ganglion cells. The method will prove useful for the molecular characterization of adult retinal ganglion cells.

Animals↗
PubMed2002

Neuronal organization and cell interactions of the cat stellate ganglion.

The functional structure of the cat stellate ganglion (SG) and, in particular, its extra- and intraganglionic connections and neuronal organization, were investigated using histochemical, immunohistochemical, morphological and histological methods. Retrograde axonal transport of horseradish peroxidase was used to determine most of the extraganglionic interactions. Of the targets tested, the most extensive efferent connections of the SG were with the stemocleidomastoid muscle, trachea, esophagus and heart. Neurons of the SG also send a small number of postganglionic efferents to the thyroid and stomach. Furthermore, ganglion cells send axons to the spinal ganglia. Several afferent connections of the SG were determined. Sympathetic preganglionic neurons of segments C8-T10 of the spinal cord, sensory neurons of C8-T9 spinal ganglia, intramural ganglia of the thoracic viscera and the reticular formation of the medulla oblongata send their axons to the SG. Intraganglionic interactions of intemeurons with principal ganglionic cells were assumed to occur, based on the presence of interneurons immunoreactive to GABA and substance P. GABA- and substance P-immunoreactive fibers located around a small number of postganglionic neurons were also identified. Morphological study revealed asymmetry between the left and right ganglia: the right ganglion is larger than the left and contains more cells. This asymmetry was also reflected in basic structural parameters of neurons, such as average neuronal area and average diameter of cell somata. The present data has been used to develop a scheme for the basic inter- and intraneuronal connections of the cat SG. This ganglion is a true nervous center, with postganglionic neurons, some of which might be performing sensory functions, and interneurons. The ganglion is connected not only with the spinal cord and spinal ganglia, but also with neurons of the intramural ganglia and, by direct links, with efferent neurons of the medulla oblongata. Thus, the SG may play an essential role in viscera-visceral reflexes.

Afferent Pathways↗
PubMed1990

Neuronal structure of the lacertilian parietal eye, I: A retrograde label and electron-microscopic study of the ganglion cells in the photoreceptor layer.

The cellular connectivity of the lacertilian parietal eye is not well understood. Because the intercellular connections establish the foundation for information processing, we have investigated cellular connectivity of one cell type in this simple vertebrate retina. We also developed an in vitro preparation to study the anatomy of the parietal eye visual system. Horseradish peroxidase transport in the in vitro preparation revealed a class of displaced ganglion cells occupying positions among the photoreceptors, in a location where the presence of interneurons had been suggested. Three-dimensional reconstruction at the electron-microscopic level showed that the morphology and synaptic input of these displaced ganglion cells is different from that of the previously known ganglion cells. The displaced ganglion cells receive an average of about 13 ribbon synapses from photoreceptors. The ribbon input is equally distributed between the soma and dendritic arbor. Junctional membrane measurement and ethanolic phosphotungstic acid-staining provided evidence for the existence of non-ribbon synaptic contacts (synaptoid junctions). Displaced ganglion cells make about 20 synaptoid junctions, 65% of which are on the dendritic arbor. The morphology of the displaced ganglion cell is such that a significant measure of synaptic input to the dendritic arbor will be transmitted to the soma.

Animals↗
PubMed1992

Variability of responses of cat retinal ganglion cells.

Previous studies of the variability of firing of retinal ganglion cells have led to apparently contradictory conclusions. To a first approximation, the variance of rate of maintained discharges of ganglion cells in cat is independent of the mean firing rate. On the other hand, the variability of responses to abrupt changes in lighting of ganglion cells in goldfish increases with increasing firing rate. To examine whether the difference is due to differences between species, we examined the variability of responses of cat ganglion cells, and find it similar to that of goldfish ganglion cells. The variance of rate of ganglion cells is neither independent of mean rate, as might be expected from maintained discharges, nor directly proportional to the mean rate, as it is for cat cortical cells. Rather, there is a nonlinear relationship between variance of rate and mean rate.

Animals↗
PubMed1992

Ganglion cell types of the turtle retina that project to the optic tectum: Intracellular HRP injections of retrogradely, rhodamine-marked cell bodies.

The turtle retina has been shown to have a variety of different morphological ganglion cell types as well as distinct physiological ganglion cell types. The major projection of the retina to the brain in nonmammalian vertebrates is to the optic tectum. In this study, we address the question of which retinal ganglion cell types project to the optic tectum in the turtle. Fluorescent rhodamine-labeled microspheres were used to trace the retinal ganglion cell projection to the superficial layers of the optic tectum. The fluorescent ganglion cell somata, retrogradely marked by transport from the contralateral optic tectum, were impaled with micropipettes containing rhodamine-horseradish peroxidase solution and this dye was iontophoresed into the cells under visual control. Most of the morphological ganglion cell types described in Golgi studies (Kolb, 1982; Kolb et al., 1988) were stained. Thus, the small cell types G1, G2, G3, G5, G6, and G7; the medium-sized types G10, G11, G12, G13, and G14; and the large-sized types G15, G16, G19, G20, and G21 project to the optic tectum in the turtle. We have added a new type, G2a, which proves to have some differences from the original G2 in branching pattern. We were unable to stain the small type G4, the medium-sized types G8 and G9, and the large cell types G17 and G18: this suggests that they might not project to the superficial layers of the dorsolateral optic tectum, at least, in the turtle.

Animals↗
PubMed1992

Visual effects of damage to P ganglion cells in macaques.

Four indices of visual performance were measured in control macaques and in macaques that had been exposed to monomeric acrylamide, a neurotoxicant that preferentially damages P retinal ganglion cells. Morphological examination of the retina and visual pathways of these monkeys showed virtually complete loss of P ganglion cells over a region extending to at least 40 deg from the fovea, and relative sparing of M ganglion cells. The four tests examined visual functions for which the visual pathway from P ganglion cells might be of great importance: visual acuity, contrast discrimination, hyperacuity, and shape discrimination. In the acrylamide-dosed monkeys, visual acuity was reduced slightly more than fourfold, a somewhat larger reduction than that seen previously after ibotenic-acid lesions of the P pathway in the geniculate. The residual acuity was in good agreement with the Nyquist frequency calculated from the density of ON or OFF M ganglion cells. Contrast increment thresholds were elevated for the dosed monkeys only in one of the two conditions tested. The elevation was found only under those spatiotemporal conditions for which we have previously shown that contrast thresholds are increased by acrylamide exposure, and was most marked at low background contrasts. Vernier acuity was elevated in one dosed monkey, but not affected in a second monkey that also had severe loss of P ganglion cells. Finally, we found no effect of acrylamide exposure on the number of training trials required to learn simple or complex shape discriminations. These results support previous findings in showing that the P pathway mediates visual acuity, and they show that several other important aspects of visual perception are not exclusively dependent on the P pathway.

Acrylamide↗
PubMedDate not supplied

The retinal ganglion cell distribution and the representation of the visual field in area 17 of the owl monkey, Aotus trivirgatus.

The distribution of ganglion cells and displaced amacrine cells was determined in whole-mounted Aotus retinae. In contrast to diurnal simians, Aotus has only a rudimentary fovea. Ganglion cell density decreases towards the periphery at approximately the same rate along all meridians, but is 1.2-1.8 times higher in the nasal periphery when compared to temporal region at the same eccentricities. The total number of ganglion cells varied from 421,500 to 508,700. Ganglion cell density peaked at 15,000/mm2 at 0.25 mm dorsal to the fovea. The displaced amacrine cells have a shallow density gradient, their peak density in the central region is about 1500-2000/mm2 and their total number varied from 315,900 to 482,800. Comparison between ganglion cell density and areal cortical magnification factor for the primary visual cortex, area 17, shows that there is not a simple proportional representation of the ganglion cell distribution. There is an overrepresentation of the central 10 deg of the visual field in the visual cortex. The present results for Aotus and the results of a similar analysis of data from other primates indicate that the overrepresentation of the central visual field is a general feature of the visual system of primates.

Animals↗
PubMedDate not supplied

Severity of ganglion cell death during early postnatal development is modulated by both neuronal activity and binocular competition.

The influence of postnatal neuronal activity on the magnitude of retinal ganglion cell death has been studied in cats. A constant blockade of activity in one eye starting just after birth does not change the severity of naturally occurring ganglion cell death, and as in normal animals, the ganglion cell population declines from 250,000 to 160,000 over a 4- to 6-week period. However, the population of retinal ganglion cells in the active untreated eye of monocularly deprived cats is increased 12% above normal (180,000 vs. 160,000 in each of four cases). This increase of 20,000 cells is permanent, and presumably reflects the competitive advantage in their target nuclei that the still active axons have over their silenced companions from the treated eye. Surprisingly, in one animal treated successfully for long duration with TTX in both from the population of ganglion cells was elevated in both eyes (200,000 and 208,000 ganglion cells). This increase matches that achieved by early unilateral enucleation (Williams et al., 1983). Our results demonstrate that the complete blockade of activity reduces the severity of naturally occurring cell death in a population of CNS sensory neurons. The effects of unilateral blockade emphasize that the activity-dependent modulation of neuron death only occurs under conditions that do not place the inactive population of neurons at a competitive disadvantage.

Action Potentials↗
PubMedDate not supplied

Enkephalin-immunoreactive ganglion cells in the pigeon retina.

A small number of enkephalin-like immunoreactive cells were observed in the ganglion cell layer of the pigeon retina. Many of these neurons were identified as ganglion cells, since they were retrogradely labeled after injections of fluorescent latex microspheres in the contralateral optic tectum. These ganglion cells were mainly distributed in the inferior retina, and their soma sizes ranged from 12-26 microns in the largest axis. The enkephalin-containing ganglion cells appear to represent only a very small percentage of the ganglion cells projecting to the optic tectum (less than 0.1%). Two to 7 weeks after removal of the neural retina, there was an almost complete elimination of an enkephalin-like immunoreactive plexus in layer 3 of the contralateral, rostrodorsal optic tectum. These data provide evidence for the existence of a population of enkephalinergic retinal ganglion cells with projections to the optic tectum.

Animals↗
PubMedDate not supplied

Inhibition is not required for the production of transient spiking responses from retinal ganglion cells.

Ganglion cells responding only transiently to changes in illumination are found in many different vertebrate retinas. The interactions underlying formation of these transient responses are still poorly understood. Two recently proposed hypotheses are (1) functional inhibitory pathways are necessary for transient response production, and (2) direct inhibition of the ganglion cell has little effect on its output. Here, we examine these conclusions by using cell-attached patch-clamp recordings of spiking, whole-cell recordings of synaptic currents, and computer modeling. We found that picrotoxin (a GABA(A) and GABA(C) receptor antagonist), bicuculline (a GABA(A) receptor antagonist), and strychnine (a glycine receptor antagonist), applied either singly or in combination, always failed to convert transient responses to sustained responses. Application of the GABA(B) antagonist CGP35348 in the presence of picrotoxin and strychnine also failed to convert transient responses into sustained responses. Whole-cell recordings of synaptic currents at various holding potentials indicated that direct inhibitory inputs to ganglion cells limit the duration of net excitation, implying that direct inhibition does act to truncate the ganglion cell spiking response. Computer simulations using spiking and synaptic data from combined cell-attached and whole-cell recordings supported this interpretation. We conclude that inhibitory pathways are not required for generation of transient responses, but these pathways do serve to modulate transient ganglion cell spiking responses. We find that this modulation occurs, in part, via inhibitory inputs directly to the ganglion cell.

Action Potentials↗
PubMedDate not supplied

Organization of the inner retina following early elimination of the retinal ganglion cell population: effects on cell numbers and stratification patterns.

The present study has examined the effects of early ganglion cell elimination upon the organization of the inner retina in the ferret. The population of retinal ganglion cells was removed by optic nerve transection on the second postnatal day, and retinas were subsequently studied in adulthood. Numbers of amacrine and bipolar cells were compared in the nerve-transected and nerve-intact retinas of operated ferrets, while stratification patterns within the inner plexiform layer were compared in these and in normal ferret retinas. Early ganglion cell elimination was found to produce a 25% reduction in the population of glycine transporter-immunoreactive amacrine cells, and 18 and 15% reductions in the populations of parvalbumin and calbindin-immunoreactive amacrine cells, respectively. GABAergic amacrine cells were also reduced by 34%. The number of calbindin-immunoreactive displaced amacrine cells, by contrast, had increased in the ganglion cell-depleted retina, being three times their normal number. Other amacrine and bipolar cell types were unaffected. Despite these changes, the stratification patterns associated with these cell types remained largely intact within the inner plexiform layer. The present results demonstrate a class-specific dependency of inner retinal neurons upon the ganglion cell population in early postnatal life, but the ganglion cells do not appear to provide any critical signals for stratification within the inner plexiform layer, at least not after birth. Since they themselves do not produce stratified dendritic arbors until well after birth, the signals for stratification of the bipolar and amacrine cell processes should arise from other sources.

Amino Acid Transport Systems, Neutral↗