Research Article
Retinal bipolar cells: Temporal filtering of signals from cone photoreceptors
- DWIGHT A. BURKHARDT, PATRICK K. FAHEY, MICHAEL A. SIKORA
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- 20 December 2007, pp. 765-774
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The temporal dynamics of the response of neurons in the outer retina were investigated by intracellular recording from cones, bipolar, and horizontal cells in the intact, light-adapted retina of the tiger salamander (Ambystoma tigrinum), with special emphasis on comparing the two major classes of bipolars cells, the ON depolarizing bipolars (Bd) and the OFF hyperpolarizing bipolars (Bh). Transfer functions were computed from impulse responses evoked by a brief light flash on a steady background of 20 cd/m2. Phase delays ranged from about 89 ms for cones to 170 ms for Bd cells, yielding delays relative to that of cones of about 49 ms for Bh cells and 81 ms for Bd cells. The difference between Bd and Bh cells, which may be due to a delay introduced by the second messenger G-protein pathway unique to Bd cells, was further quantified by latency measurements and responses to white noise. The amplitude transfer functions of the outer retinal neurons varied with light adaptation in qualitative agreement with results for other vertebrates and human vision. The transfer functions at 20 cd/m2 were predominantly low pass with 10-fold attenuation at about 13, 14, 9.1, and 7.7 Hz for cones, horizontal, Bh, and Bd cells, respectively. The transfer function from the cone voltage to the bipolar voltage response, as computed from the above measurements, was low pass and approximated by a cascade of three low pass RC filters (“leaky integrators”). These results for cone→bipolar transmission are surprisingly similar to recent results for rod→bipolar transmission in salamander slice preparations. These and other findings suggest that the rate of vesicle replenishment rather than the rate of release may be a common factor shaping synaptic signal transmission from rods and cones to bipolar cells.
When S-cones contribute to chromatic global motion processing
- ALEXA I. RUPPERTSBERG, SOPHIE M. WUERGER, MARCO BERTAMINI
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- 11 April 2007, pp. 1-8
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There is common consensus now that color-defined motion can be perceived by the human visual system. For global motion integration tasks based on isoluminant random dot kinematograms conflicting evidence exists, whether observers can (Ruppertsberg et al., 2003) or cannot (Bilodeau & Faubert, 1999) extract a common motion direction for stimuli modulated along the isoluminant red-green axis. Here we report conditions, in which S-cones contribute to chromatic global motion processing. When the display included extra-foveal regions, the individual elements were large (∼0.3°) and the displacement was large (∼1°), stimuli modulated along the yellowish-violet axis proved to be effective in a global motion task. The color contrast thresholds for detection for both color axes were well below the contrasts required for global motion integration, and therefore the discrimination-to-detection ratio was >1. We conclude that there is significant S-cone input to chromatic global motion processing and the extraction of global motion is not mediated by the same mechanism as simple detection. Whether the koniocellular or the magnocellular pathway is involved in transmitting S-cone signals is a topic of current debate (Chatterjee & Callaway, 2002).
EDITORIAL
General editorial
- RON DOUGLAS
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- 26 June 2007, p. 231
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In the past two years the field of fish vision research has lost three central figures, Bill McFarland, Bill Muntz, and Joe Bilotta. This volume of Visual Neuroscience is dedicated to them. Since the inception of this volume, Henk Spekreijse and Adam Locket, who made significant contributions to fish vision, have also passed away; they are also remembered.
Research Article
Local influence of mitochondrial calcium transport in retinal amacrine cells
- MADHUMITA SEN, EMILY MCMAINS, EVANNA GLEASON
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- 16 August 2007, pp. 663-678
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Ca2+-dependent synaptic transmission from retinal amacrine cells is thought to be initiated locally at dendritic processes. Hence, understanding the spatial and temporal impact of Ca2+ transport is fundamental to understanding how amacrine cells operate. Here, we provide the first examination of the local effects of mitochondrial Ca2+ transport in neuronal processes. By combining mitochondrial localization with measurements of cytosolic Ca2+, the local impacts of mitochondrial Ca2+ transport for two types of Ca2+ signals were investigated. Disruption of mitochondrial Ca2+ uptake with carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) produces cytosolic Ca2+ elevations. The amplitudes of these elevations decline with distance from mitochondria suggesting that they are related to mitochondrial Ca2+ transport. The time course of the FCCP-dependent Ca2+ elevations depend on the availability of ER Ca2+ and we provide evidence that Ca2+ is released primarily via nearby ryanodine receptors. These results indicate that interactions between the ER and mitochondria influence cytosolic Ca2+ in amacrine cell processes and cell bodies. We also demonstrate that the durations of glutamate-dependent Ca2+ elevations are dependent on their proximity to mitochondria in amacrine cell processes. Consistent with this observation, disruption of mitochondrial Ca2+ transport alters the duration of glutamate-dependent Ca2+ elevations near mitochondria but not at sites more than 10 μm away. These results indicate that mitochondria influence local Ca2+-dependent signaling in amacrine cell processes.
Alteration of retinal intrinsic survival signal and effect of α2–adrenergic receptor agonist in the retina of the chronic ocular hypertension rat
- HWA SUN KIM, YONG IK CHANG, JIE HYUN KIM, CHAN KEE PARK
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- 19 July 2007, pp. 127-139
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The purpose of this study is to examine the retinal expression of intrinsic cell survival molecules and to elucidate the effect of an α2-adrenergic receptor agonist in the chronic ocular hypertensive rat model. Chronic ocular hypertension was induced in both eyes of each rat by episcleral vein cauterization. Two five-microliter drops of the selective α2-adrenoceptor agonist brimonidine 0.2% (Alphagan; Allergan Inc., Irvine, CA, USA) were topically administered twice daily for up to eight weeks in one eye. The fellow eye received balanced salt solution as a control. Protein and mRNA expression were evaluated at 1, 4, and 8 weeks after injury. Retinal expression of BDNF, Akt, and GFAP was assessed using immunohistochemistry. Retinal levels of mRNA for BDNF, bcl-2, and bcl-xL were determined using semi-quantitative RT-PCR. Retinal ganglion cell (RGC) density was evaluated after retrograde labeling with 4-Di-10-ASP (DiA). A significant decrease in RGC density was observed in ocular hypertensive eyes. Cauterized eyes showed an increase in GFAP expression from one week after injury, and the expression of bcl-2, bcl-xL, and BDNF mRNA was also increased. Treatment of ocular hypertensive eyes with brimonidine resulted in a reduction in RGC loss, a decrease in the level of GFAP immunoreactivity, and an increment in BDNF mRNA and p-Akt expression. Brimonidine appears to protect RGCs from neurodegeneration through mechanisms involving α2-adrenergic receptor mediated survival signal activation and up-regulation of endogenous neurotrophic factor expression in the chronic ocular hypertensive rat retina.
IN MEMORIAM
In memoriam
- ROBERT F. MILLER, ELIO RAVIOLA
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- 28 September 2007, pp. 445-447
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The vision research community lost a valuable colleague and friend when Ramon (Ray) Dacheux II died on May 30, 2006 at his home in Birmingham, Alabama. Ray was 58 and had been ill with a brain tumor for two years.
MEMORIALS
A tribute to Joe Bilotta
- PAUL DEMARCO, ELIZABETH LEMERISE
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- 29 May 2007, p. 233
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Dr. Joseph Bilotta, an eminent scientist in the field of fish visual neurophysiology and psychophysics, died suddenly and unexpectedly on January 2, 2006. A native of Niagara Falls, NY, Joe was born October 21, 1955. His passion for learning took him on a journey from an Associate Degree in Mathematics in 1975 from Niagara County Community College to a Ph.D. in Experimental Psychology conferred by the City University of New York in 1987 under the mentorship of Dr. Israel Abramov. Joe continued his training at Vanderbilt University, working as a post-doctoral fellow in the laboratory of Dr. Maureen Powers. In 1991, Joe joined the faculty of Western Kentucky University as an assistant professor of Psychology, and quickly moved through the ranks, becoming full professor in 2001.
Research Article
Test of the paired-flash electroretinographic method in mice lacking b-waves
- JENNIFER J. KANG DERWENT, SHANNON M. SASZIK, HIDETAKA MAEDA, DEBORAH M. LITTLE, MACHELLE T. PARDUE, LAURA J. FRISHMAN, DAVID R. PEPPERBERG
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- 19 July 2007, pp. 141-149
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Previous studies of rod photoreceptors in vivo have employed a paired-flash electroretinographic (ERG) technique to determine rod response properties. To test whether absence versus presence of the ERG b-wave affects the photoreceptor response derived by the paired-flash method, we examined paired-flash-derived responses obtained from nob mice, a mutant strain with a defect in signal transduction between photoreceptors and ON bipolar cells that causes a lack of the b-wave. Normal littermates of the nob mice served as controls. The normalized amplitude-intensity relation of the derived response determined in nob mice at the near-peak time of 86 ms was similar to that determined for the controls. The full time course of the derived rod response was obtained for test flash strengths ranging from 0.11 to 17.38 scotopic cd s m−2 (sc cd s m−2). Time-course data obtained from nob and control mice exhibited significant but generally modest differences. With saturating test flash strengths, half-recovery times for the derived response of nobversus control mice differed by ∼60 ms or less about the combined (nob and control) average respective values. Time course data also were obtained before versus after intravitreal injection of l-2-amino-4-phosphonobutyrate (APB) (which blocks transmission from photoreceptors to depolarizing bipolar cells) and of cis 2,3-piperidine dicarboxylic acid (PDA) (which blocks transmission to OFF bipolar cells, and to horizontal, amacrine and ganglion cells). Neither APB nor PDA substantially affected derived responses obtained from nob or control mice. The results provide quantitative information on the effect of b-wave removal on the paired-flash-derived response in mouse. They argue against a substantial skewing effect of the b-wave on the paired-flash-derived response obtained in normal mice and are consistent with the notion that, to good approximation, this derived response represents the isolated flash response of the photoreceptors in both nob and normal mice.
Temporal properties of surround suppression in cat primary visual cortex
- SÉVERINE DURAND, TOBE C.B. FREEMAN, MATTEO CARANDINI
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- 09 August 2007, pp. 679-690
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The responses of neurons in primary visual cortex (V1) are suppressed by stimuli presented in the region surrounding the receptive field. There is debate as to whether this surround suppression is due to intracortical inhibition, is inherited from lateral geniculate nucleus (LGN), or is due to a combination of these factors. The mechanisms involved in surround suppression may differ from those involved in suppression within the receptive field, which is called cross-orientation suppression. To compare surround suppression to cross-orientation suppression, and to help elucidate its underlying mechanisms, we studied its temporal properties in anesthetized and paralyzed cats. We first measured the temporal resolution of suppression. While cat LGN neurons respond vigorously to drift rates up to 30 Hz, most cat V1 neurons stop responding above 10–15 Hz. If suppression originated in cortical responses, therefore, it should disappear above such drift rates. In a majority of cells, surround suppression decreased substantially when surround drift rate was above ∼15 Hz, but some neurons demonstrated suppression with surround drift rates as high as 21 Hz. We then measured the susceptibility of suppression to contrast adaptation. Contrast adaptation reduces responses of cortical neurons much more than those of LGN neurons. If suppression originated in cortical responses, therefore, it should be reduced by adaptation. Consistent with this hypothesis, we found that prolonged exposure to the surround stimulus decreased the strength of surround suppression. The results of both experiments differ markedly from those previously obtained in a study of cross-orientation suppression, whose temporal properties were found to resemble those of LGN neurons. Our results provide further evidence that these two forms of suppression are due to different mechanisms. Surround suppression can be explained by a mixture of thalamic and cortical influences. It could also arise entirely from intracortical inhibition, but only if inhibitory neurons respond to somewhat higher drift rates than most cortical cells.
Evidence that L-AP5 and D,L-AP4 can preferentially block cone signals in the rat retina
- DANIEL G. GREEN, NATALIA V. KAPOUSTA-BRUNEAU
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- 11 April 2007, pp. 9-15
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Several lines of evidence suggest that, as concentrations of two agonists of group III metabotropic glutamate receptors are increased, cone contributions to the b-wave are blocked before rod contributions. Application of L-AP5 (L-2-amino-5-phosphonobutyric acid) at concentrations of 50 μM and D,L-AP4 (D,L-2-amino-4-phosphonobutyric acid) at concentrations 2 μM had a greater effect in reducing the amplitude of the rat ERG b-wave at high light intensities than at low light intensities. The amplitude reduction occurs at flash intensities that saturate rod photoreceptor responses. When steady backgrounds are used to saturate rod photoreceptors, the b-wave responses show increased long-wavelength sensitivity. Responses on a rod saturating background are blocked by adding L-AP5 or AP4 at the above concentrations to the perfusate. Further evidence for metabotrophic receptors being involved comes from the observation that even when ionotropic glutamate receptors are pharmacologically blocked with MK801 and DNQX, AP4 selectively blocks cone contributions to the b-wave. Thus we suggest that the type III metabotrophic receptors on depolarizing cone bipolar cells or cone synaptic terminals are affected by concentrations of L-AP5 and D,L-AP4 that have minimal effects on rod bipolar cells or rod synaptic terminals.
Functional polarity of dendrites and axons of primate A1 amacrine cells
- CHRISTOPHER M. DAVENPORT, PETER B. DETWILER, DENNIS M. DACEY
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- 29 May 2007, pp. 449-457
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The A1 cell is an axon-bearing amacrine cell of the primate retina with a diffusely stratified, moderately branched dendritic tree (∼400 μm diameter). Axons arise from proximal dendrites forming a second concentric, larger arborization (>4 mm diameter) of thin processes with bouton-like swellings along their length. A1 cells are ON-OFF transient cells that fire a brief high frequency burst of action potentials in response to light (Stafford & Dacey, 1997). It has been hypothesized that A1 cells receive local input to their dendrites, with action potentials propagating output via the axons across the retina, serving a global inhibitory function. To explore this hypothesis we recorded intracellularly from A1 cells in an in vitro macaque monkey retina preparation. A1 cells have an antagonistic center-surround receptive field structure for the ON and OFF components of the light response. Blocking the ON pathway with L-AP4 eliminated ON center responses but not OFF center responses or ON or OFF surround responses. Blocking GABAergic inhibition with picrotoxin increased response amplitudes without affecting receptive field structure. TTX abolished action potentials, with little effect on the sub-threshold light response or basic receptive field structure. We also used multi-photon laser scanning microscopy to record light-induced calcium transients in morphologically identified dendrites and axons of A1 cells. TTX completely abolished such calcium transients in the axons but not in the dendrites. Together these results support the current model of A1 function, whereby the dendritic tree receives synaptic input that determines the center-surround receptive field; and action potentials arise in the axons, which propagate away from the dendritic field across the retina.
Cytoskeleton alteration correlates with gross structural plasticity in the cat lateral geniculate nucleus
- MATTHEW R. KUTCHER, KEVIN R. DUFFY
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- 04 October 2007, pp. 775-785
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Monocular deprivation during early development causes rearrangement of neural connections within the visual cortex that produces a shift in ocular dominance favoring the non-deprived eye. This alteration is manifested anatomically within deprived layers of the lateral geniculate nucleus (LGN) where neurons have smaller somata and reduced geniculocortical terminal fields compared to non-deprived counterparts. Experiments using monocular deprivation have demonstrated a spatial correlation between cytoskeleton alteration and morphological change within the cat LGN, raising the possibility that subcellular events mediating deprivation-related structural rearrangement include modification to the neuronal cytoskeleton. In the current study we compared the spatial and temporal relationships between cytoskeleton alteration and morphological change in the cat LGN. Cross-sectional soma area and neurofilament labeling were examined in the LGN of kittens monocularly deprived at the peak of the critical period for durations that ranged from 1 day to 7 months. After 4 days of deprivation, neuron somata within deprived layers of the LGN were significantly smaller than those within non-deprived layers. This structural change was accompanied by a spatially coincident reduction in neurofilament immunopositive neurons that was likewise significant after 4 days of deprivation. Both anatomical effects reached close to their maximum by 10 days of deprivation. Results from this study demonstrate that alteration to the neuronal cytoskeleton is both spatially and temporally linked to the gross structural changes induced by monocular deprivation.
Naturalistic color discriminations in polymorphic platyrrhine monkeys: Effects of stimulus luminance and duration examined with functional substitution
- MICKEY P. ROWE, GERALD H. JACOBS
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- 11 April 2007, pp. 17-23
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X-linked photopigment polymorphism produces six different color vision phenotypes in most species of New World monkey. In the subfamily Callitrichinae, the three M/L alleles underlying these different phenotypes are present at unequal frequencies suggesting that selective pressures other than heterozygous-advantage operate on these alleles. Earlier we investigated this hypothesis with functional substitution, a technique using a computer monitor to simulate colors as they would appear to humans with monkey visual pigments (Visual Neuroscience21:217–222, 2004). The stimuli were derived from measurements of ecologically relevant fruit and foliage. We found that discrimination performance depended on the relative spectral positioning of the substituted M and L pigment pair. Here we have undertaken a systematic examination of two simulation parameters—test field luminance and stimulus duration. Discriminability of the fruit colors depended on which phenotype was simulated but only at short stimulus durations and/or low luminances. Under such conditions, phenotypes with the larger pigment peak separations performed better. At longer durations and higher luminances, differences in performance across different substitutions tended to disappear. The stimuli used in this experiment were analyzed with several color discrimination models. There was limited agreement among the predictions made by these models regarding the capabilities of animals with different pigment pairs and none predicted the dependence of discrimination on changes in luminance and stimulus duration.
MEMORIALS
A tribute to William N. “MAC” McFarland (1926–2004)
- ELLIS LOEW
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- 29 May 2007, p. 235
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On August 31, 2004 William N. “MAC” McFarland died in Mt. Vernon, Washington just 11 days shy of his 79th birthday. He was into his second post-retirement professorship (from Cornell and USC) at the Friday Harbor Labs of the University of Washington. Rather than the usual CV with a list of awards and accomplishments, of which Mac had many, I would like to posit the following question, “Why should Mac be honored in this issue?” To those of us who knew and worked with him, the fact that he was “Mac” says it all. However, to those who did not know him, more justification is needed.
Research Article
Projections of the nucleus of the basal optic root in pigeons (Columba livia): A comparison of the morphology and distribution of neurons with different efferent projections
- DOUGLAS R.W. WYLIE, JANELLE M.P. PAKAN, CAMERON A. ELLIOTT, DAVID J. GRAHAM, ANDREW N. IWANIUK
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- 04 October 2007, pp. 691-707
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The avian nucleus of the basal optic root (nBOR) is a visual structure involved in the optokinetic response. nBOR consists of several morphologically distinct cell types, and in the present study, we sought to determine if these different cell types had differential projections. Using retrograde tracers, we examined the morphology and distribution of nBOR neurons projecting to the vestibulocerebellum (VbC), inferior olive (IO), dorsal thalamus, the pretectal nucleus lentiformis mesencephali (LM), the contralateral nBOR, the oculomotor complex (OMC) and a group of structures along the midline of the mesencephalon. The retrogradely labeled neurons fell into two broad categories: large neurons, most of which were multipolar rather than fusiform and small neurons, which were either fusiform or multipolar. From injections into the IO, LM, contralateral nBOR, and structures along the midline-mesencephalon small nBOR neurons were labeled. Although there were no differences with respect to the size of the labeled neurons from these injections, there were some differences with the respect to the distribution of labeled neurons and the proportion of multipolar vs. fusiform neurons. From injections into the VbC, the large multipolar cells were labeled throughout nBOR. The only other cases in which these large neurons were labeled were contralateral OMC injections. To investigate if single neurons project to multiple targets we used paired injections of red and green fluorescent retrograde tracers into different targets. Double-labeled neurons were never observed indicating that nBOR neurons do not project to multiple targets. We conclude that individual nBOR neurons have unique projections, which may have differential roles in processing optic flow and controlling the optokinetic response.
Response properties of a unique subtype of wide-field amacrine cell in the rabbit retina
- STEWART A. BLOOMFIELD, BÉLA VÖLGYI
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- 29 May 2007, pp. 459-469
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We studied the morphology and physiology of a unique wide-field amacrine cell in the rabbit retina. These cells displayed a stereotypic dendritic morphology consisting of a large, circular and monostratified arbor that often extended over 2 mm. Their responses contained both somatic and dendritic sodium spikes suggesting active propagation of synaptic signals within the dendritic arbor. This idea is supported by the enormous size of their ON-OFF receptive fields. Interestingly, these cells exhibited separate ON and OFF receptive fields that, while concentric, were vastly different in size. Whereas the ON receptive field of these cells extended nearly 2 mm, the OFF receptive field was typically 75% smaller. Blockade of voltage-gated sodium channels with QX-314 dramatically reduced the large ON receptive field, but had little effect on the smaller OFF receptive field. These results indicate a spatial disparity in the location of on- and off-center bipolar cell inputs to the dendritic arbor of wide-field amacrine cells. In addition, the active propagation of signals suggests that synaptic inputs are integrated both locally and globally within the dendritic arbor.
Temporal resolution and temporal transfer properties: Gabaergic and cholinergic mechanisms
- KONSTANTIN BEHREND, BORIS BENKNER, CARLOS MORA-FERRER
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- 20 December 2007, pp. 787-797
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Temporal resolution is a basic property of the visual system and critically depends upon retinal temporal coding properties which are also of importance for directional coding. Whether the temporal coding properties for directional coding derive form inherent properties or critically depend upon the temporal coding mechanisms is unclear. Here, the influence of acetylcholine and GABA upon photopic temporal coding was investigated in goldfish, using flicker stimuli, in a behavioral and an electrophysiological (ERG) approach. The goldfish temporal resolution ability decreased from more than 90% correct choices at 20 Hz flicker frequency to about 65% at 45 Hz flicker frequency with a flicker fusion frequency of approximately 39 Hz. Blockade of GABAa-receptors reduced the flicker fusion frequency to about 23 Hz, not affecting temporal resolution below 20 Hz flicker frequency. Partial blockade of nicotinic acetylcholine receptors reduced the flicker fusion frequency slightly and lowered the temporal resolution ability in the 25–30 Hz range. Blockade of muscarinic acetylcholine receptors had a smaller effect than the partial blockade of nicotinic acetylcholine receptors. In ERG-recordings, blocking GABAa-receptors increased the a- and b-wave amplitude, induced a delay, an increase and a slow fall-off of the d-wave. Blocking GABAc-receptors had little effect. Blocking GABAa- or GABAa/c-receptors changed the temporal resolution, when expressed as a linear filter, from a 3rd degree filter with resonance to a low order low-pass filter with a low upper limit frequency. The temporal transfer properties were barely changed by blocking either nicotinic or muscarinic acteylcholine receptors, although ERG-components increased in amplitude to varying degrees. The behavioral and electrophysiological data indicate the important role of GABA for temporal processing but little involvement of the cholinergic system. It is proposed that the interaction of the GABAergic amacrine cell network and bipolar cells determines the gain of the retinal temporal coding in the upper frequency range.
The ageing photoreceptor
- ALEXANDER CUNEA, GLEN JEFFERY
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- 19 July 2007, pp. 151-155
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With age many retinal neurons are lost. In humans the rod photoreceptor population in the perimacular region is subject to approximately 30% loss over life. Those that remain have been reported to suffer from extensive convolutions and localized swellings of their outer segments abnormally increasing their disc content and outer segment length. Here we examine quantitatively age-related changes in rat rod photoreceptors. The rat retina is ∼97% rod dominated. Here, aged rods showed significant reductions in outer segment length. The discs in their outer segments had a similar density, irrespective of whether they were young or old, however, in aged animals a higher proportion were misregistered. Surprisingly, in all of the tissue examined, we found no evidence for any convolution of outer segments or localized swelling as reported in humans, rather all remained straight. There are methodological differences between the research reported here and that undertaken on human retinae. There are also major differences in overall retinal architecture between humans and rodents that could contribute to differences in the aging process of individual cells. If it is the case that individual photoreceptors age differently in rodents compared to humans, it may pose significant problems for the use of this animal model in studies of ageing and age related outer retinal disease.
Glycine receptors of A-type ganglion cells of the mouse retina
- SRIPARNA MAJUMDAR, LIANE HEINZE, SILKE HAVERKAMP, ELENA IVANOVA, HEINZ WÄSSLE
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- 29 May 2007, pp. 471-487
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A-type ganglion cells of the mouse retina represent the visual channel that transfers temporal changes of the outside world very fast and with high fidelity. In this study we combined anatomical and physiological methods in order to study the glycinergic, inhibitory input of A-type ganglion cells. Immunocytochemical studies were performed in a transgenic mouse line whose ganglion cells express green fluorescent protein (GFP). The cells were double labeled for GFP and the four α subunits of the glycine receptor (GlyR). It was found that most of the glycinergic input of A-type cells is through fast, α1-expressing synapses. Whole-cell currents were recorded from A-type ganglion cells in retinal whole mounts. The response to exogenous application of glycine and spontaneous inhibitory postsynaptic currents (sIPSCs) were measured. By comparing glycinergic currents recorded in wildtype mice and in mice with specific deletions of GlyRα subunits (Glra1spd-ot, Glra2−/−, Glra3−/−), the subunit composition of GlyRs of A-type ganglion cells could be further defined. Glycinergic sIPSCs of A-type ganglion cells have fast kinetics (decay time constant τ = 3.9 ± 2.5 ms, mean ± SD). Glycinergic sIPSCs recorded in Glra2−/− and Glra3−/− mice did not differ from those of wildtype mice. However, the number of glycinergic sIPSCs was significantly reduced in Glra1spd-ot mice and the remaining sIPSCs had slower kinetics than in wildtype mice. The results show that A-type ganglion cells receive preferentially kinetically fast glycinergic inputs, mediated by GlyRs composed of α1 and β subunits.
Differential distribution of glycine transporters in Müller cells and neurons in amphibian retinas
- ZHENG JIANG, BAOQIN LI, FRANTISEK JURSKY, WEN SHEN
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- 19 July 2007, pp. 157-168
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Amphibian retinas are commonly used for electrophysiological studies on neural function and transduction because they share the same general properties as higher vertebrate retinas. Glycinergic synapses have been well described in amphibian retinas. However, the role of glycine transporters in the synapses is largely unknown. We studied the distribution and function of glycine transporters in the retinas from tiger salamanders, mudpuppies, and leopard frogs by immunofluorescence labeling and whole-cell recording methods. Our results indicated that GlyT1- and GlyT2-like transporters were present in Müller cells and neurons, respectively. GlyT1 labeling was present in Müller glial cells and co-localized with Glial fibrillary acidic protein (GFAP), a Müller cell marker, whereas the GlyT2 immunoreactivity was present in the somas of amacrine cells (ACs) and processes in the inner plexiform layer (IPL) and the outer plexiform layer (OPL). Because the axon processes of glycinergic interplexiform cells (IPCs) are the only source of glycine input in the OPL, GlyT2 staining revealed a spatial pattern of the axon processes of IPCs in the OPL. The function of GlyT2 in the IPCs was studied in tiger salamander retinal horizontal cells (HCs) by whole-cell gramicidin perforated recording. The results demonstrated that inhibition of GlyT2 by a specific inhibitor, amoxapine, increased a tonic glycine input to HCs. Thus, the GlyT2 transporter is responsible for uptake of synaptic glycine in the outer retina. We also compared the distribution of glycine transporters in other amphibian species: salamander, mudpuppy, and frog. The results are consistent with the general pattern that GlyT1-like transporters are present in Müller cells and GlyT2-like transporters in neurons in amphibian retinas.