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Results for “Retinal Cone Photoreceptor Cells”

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PubMed1995

ERG OFF response in frog retina: light adaptation and effect of 2-amino-4-phosphonobutyrate.

The intensity-response (V/log I) function of ERG OFF response (d-wave) in dark and light adapted superfused frog eyecups was investigated before and after blockade of the retinal ON channel by 2-amino-4-phosphonobutyrate (APB). The V/log I function of the dark adapted d-wave had two distinct components, each of them consisting of an ascendent and descendent part. In eyes adapted to mesopic or photopic background the V/log I function had only one component. It was shifted to the right along the intensity axis, had a steeper slope and a higher maximal response amplitude compared with the two components of the dark-adapted V/log I curve. Perfusion with 200 mumol APB markedly increased the d-wave amplitude at all stimulus intensities except for the threshold ones in both dark and light adapted eyes. The position of the V/log I curve was shifted slightly to the left along the intensity axis in dark adapted eyes, but was not changed in light adapted eyes. Thus the adaptational mechanism responsible for changes in the decremental sensitivity with increased background illumination was not altered by APB. The effect of APB was studied also in chromatically adapted eyes, in which the responses were predominantly mediated by one photoreceptor type. The results showed that the potentiating effect of APB on d-wave did not depend on photoreceptor input.

Adaptation, Ocular↗
PubMed2003

Partial characterization of retina-derived cone neuroprotection in two culture models of photoreceptor degeneration.

PURPOSE: To define the nature and estimate the molecular weight range of soluble endogenous retinal trophic activities on cone photoreceptor survival in two models of cone degeneration. METHODS: Diffusible factors from dissociated retinal cell cultures of 8-day normal-sighted (C57BL/6J) mice were tested for cone-survival-promoting activity by two approaches and by using two independent photoreceptor degeneration models. In the first approach, mouse retinal cells were cultured on semi-permeable membranes apposed to dissociated cultures of chick embryo retina. In the second approach, conditioned medium was collected from normal mouse retinal cultures and added to embryonic chicken retina cultures or to retinal explants obtained from 5-week retinal degeneration (rd1) mice. In some experiments, conditioned medium was heated or sequentially fractionated in dialysis tubing with molecular weight cutoffs of 8, 15, and 25 kDa. The number of chicken cones and viability were determined by using morphologic criteria, colorimetric assays, and labeling with antibodies raised against visinin. Mouse cones were counted by differential double immunolabeling with antibodies against rhodopsin (rods) and arrestin (rods and cones). RESULTS: . Coculturing with normal mouse retinal cells delayed cone loss in dispersed embryonic chicken retina, by a maximum of 50% relative to the control. Conditioned medium derived from normal mouse retinas also significantly delayed cone loss in chicken cone cultures by a maximum of 1300%, compared with the control, and 40% in rd1 mouse retinal explant cultures. The survival activity in conditioned medium was destroyed by heat denaturation, and was partially retained by dialysis with a molecular weight cutoff of 25 kDa in both models. CONCLUSIONS: These strategies have identified cone-survival-stimulating activities in normal mouse retina, capable of acting across species and enhancing both structural protection and viability. Such molecules may represent candidates for clinical treatment of inherited retinal degeneration.

Animals↗
PubMed1996

Photoreceptor loss in age-related macular degeneration.

PURPOSE: The authors showed previously that parafoveal rods, but not cones, decrease during the course of adulthood in donor eyes that were screened to exclude the grossly visible macular drusen and pigmentary disturbances typical of age-related macular degeneration (AMD). Because AMD begins in the parafovea, this selective loss of rods actually may be subclinical AMD not yet visible in the fundus. If so, AMD must have a predilection for rods over cones. The authors tested this hypothesis by determining the relative numbers of cones and rods in donor eyes with mid-to late-stage AMD and in age-matched controls. METHODS: Thirteen eyes (from seven donors) with grossly visible macular drusen and pigmentary disturbances were either wholemounted for photoreceptor counts or sectioned through the fovea for histopathology and carbonic anhydrase histochemistry to label red-green cones. Eyes were assigned to AMD or control groups on the basis of histopathology and clinical history. RESULTS: Five nonexudative AMD (NE-AMD) eyes from three donors showed sparing of foveal cones and loss of rods and cones in the parafovea. In two donors, rod loss exceeded cone loss at most parafoveal locations, and in one donor, rod density was normal and cone density was reduced. In eight exudative AMD (EX-AMD) eyes from five donors, photoreceptors surviving along the margins of and overlying disciform scars were largely cones. CONCLUSIONS: Photoreceptors are lost in NE-AMD as well as in the more severe exudative form, consistent with functional and clinical studies. The authors propose that rods die in older eyes without evidence of overt retinal pigment epithelial disease. In persons susceptible to AMD, the retinal pigment epithelium becomes dysfunctional. Secondarily, rod loss continues and cones begin to degenerate. Eventually, only degenerate cones remain; ultimately, all photoreceptors may disappear.

Aged↗
PubMed2005

The nitric oxide-cGMP signaling pathway differentially regulates presynaptic structural plasticity in cone and rod cells.

Although abundant structural plasticity in the form of axonal retraction, neurite extension, and formation of presynaptic varicosities is displayed by photoreceptors after retinal detachment and during genetic and age-related retinal degeneration, the mechanisms involved are mostly unknown. We demonstrated recently that Ca(2+) influx through cGMP-gated channels in cones and voltage-gated L-type channels in rods is required for neurite extension in vitro (Zhang and Townes-Anderson, 2002). Here, we report that the nitric oxide (NO)-cGMP signaling pathway is active in photoreceptors and that its manipulation differentially regulates the structural plasticity of cone and rod cells. The NO receptor soluble guanylyl cyclase (sGC) was detected immunocytochemically in both cone and rod cells. Stimulation of sGC increased cGMP production in retinal cultures. In cone cells, quantitative analysis showed that NO or cGMP stimulated neuritic sprouting; this stimulatory effect was dependent on both Ca2+ influx through cGMP-gated channels and phosphorylation by protein kinase G (PKG). At the highest levels of cGMP, however, cone outgrowth was no longer increased. In rod photoreceptors, NO or cGMP consistently inhibited neuritic growth in a dose-dependent manner; this inhibitory effect required PKG. When NO-cGMP signaling was inhibited, changes in the neuritic development of cone and rod cells were also observed but in the opposite direction. These results expand the role of cGMP in axonal activity to adult neuritogenesis and suggest an explanation for the neurite sprouting observed in an autosomal recessive form of retinitis pigmentosa that is characterized by high cGMP levels in photoreceptor layers.

Ambystoma↗
PubMed2000

Parallel pathways for spectral coding in primate retina.

The primate retina is an exciting focus in neuroscience, where recent data from molecular genetics, adaptive optics, anatomy, and physiology, together with measures of human visual performance, are converging to provide new insights into the retinal origins of color vision. Trichromatic color vision begins when the image is sampled by short- (S), middle- (M) and long- (L) wavelength-sensitive cone photoreceptors. Diverse retinal cell types combine the cone signals to create separate luminance, red-green, and blue-yellow pathways. Each pathway is associated with distinctive retinal architectures. Thus a blue-yellow pathway originates in a bistratified ganglion cell type and associated interneurons that combine excitation from S cones and inhibition from L and M cones. By contrast, a red-green pathway, in which signals from L and M cones are opposed, is associated with the specialized anatomy of the primate fovea, in which the "midget" ganglion cells receive dominant excitatory input from a single L or M cone.

Animals↗
PubMed2000

Cloning and immunocytochemical localization of a cyclic nucleotide-gated channel alpha-subunit to all cone photoreceptors in the mouse retina.

Cyclic nucleotide-gated channels (CNGC) are ligand-gated ion channels that open and close in response to changes in the intracellular concentration of the second messengers, 3;,5;-cyclic adenosine monophosphate and 3;,5;-cyclic guanosine monophosphate. Most notably, they transduce the chemical signal produced by the absorption of light in photoreceptors into a membrane potential change, which is then transmitted to the ascending visual pathway. CNGCs have also been implicated in the signal transduction of other neurons downstream of the photoreceptors, in particular the ON-bipolar cells, as well as in other areas of the central nervous system. We therefore undertook a search for additional cyclic nucleotide-gated channels expressed in the retina. Following a degenerate reverse transcription polymerase chain reaction approach to amplify low-copy number messages, a cDNA encoding a new splice variant of CNGC alpha-subunit was isolated from mouse retina and classified as mCNG3. An antiserum raised against the carboxy-terminal sequence identified the retinal cell type expressing mCNG3 as cone photoreceptors. Preembedding immunoelectron microscopy demonstrated its membrane localization in the outer segments, consistent with its role in phototransduction. Double-labeling experiments with cone-specific markers indicated that all cone photoreceptors in the murid retina use the same or a highly conserved cyclic nucleotide-gated channel. Therefore, defects in this channel would be predicted to severely impair photopic vision.

Amino Acid Sequence↗
PubMed2001

Cones regenerate from retinal stem cells sequestered in the inner nuclear layer of adult goldfish retina.

PURPOSE: To determine whether retinal progenitor cells in the inner nuclear layer give rise to regenerated cones after laser ablation of photoreceptors in adult goldfish retina. METHODS: Using a technique developed previously in this laboratory, photoreceptors in the retina of adult goldfish were ablated with an argon laser. The mitotic marker, bromodeoxyuridine, was used to label proliferating and regenerated cells, which were identified with cell-specific markers. RESULTS: Cells proliferating locally within lesion included microglia, Müller glia, and retinal progenitors in the inner nuclear layer (INL). The nuclei of both Müller glia and associated retinal progenitors migrated from the inner to the outer nuclear layer. The proliferating retinal progenitors, which express Notch-3 and N-cadherin, regenerated cone photoreceptors and then rod photoreceptors. CONCLUSIONS: Previous work has demonstrated that photoreceptors in the goldfish retina regenerate selectively after laser ablation, but the source of regenerated cones has not been identified. The results reported here provide support for the existence of retinal stem cells within the adult fish retina that are capable of regenerating cone photoreceptors. The data also support the involvement of Müller glia in the production of regenerated cones.

Animals↗
PubMed2003

Comparative analysis and expression of CLUL1, a cone photoreceptor-specific gene.

PURPOSE: To characterize CLUL1, a cone photoreceptor-specific gene. METHODS: A comparative genomics approach was used to analyze the gene organization and protein sequence of a retinal clusterin-like protein and to identify conserved elements between human and dog. Its expression was studied by Northern and Western analyses and its localization by in situ hybridization and immunocytochemistry. RESULTS: The CLUL1 sequences of the human and dog share 85% and 73% identity, respectively, at the nucleotide and deduced amino acid level. The gene is organized into nine exons and shows strong homology, not only in exonic but also in some intronic sequences between the species. The polypeptide homology of CLUL1 to CLU, a molecular chaperone, indicates structural similarity of the two proteins. However, these data demonstrated that they present different expression profiles in the tissues, in retinal development, and in retinal diseases. Finally, CLUL1 was localized to retinal cone photoreceptor cells and a different immunolabeling in light- and dark-adapted retinas was shown. CONCLUSIONS: CLUL1 represents a potentially important gene and a candidate locus for retinal disease, particularly those diseases that affect cones.

Amino Acid Sequence↗
PubMed2000

Green cone opsin and rhodopsin regulation by CNTF and staurosporine in cultured chick photoreceptors.

PURPOSE: To investigate the regulation of visual pigment expression in chick embryo photoreceptor cells by ciliary neurotrophic factor (CNTF), and by the protein kinase inhibitor staurosporine. METHODS: Embryonic day (ED) 8 chick embryo retinal cells were dissociated and cultured at low densities for 3 days, either in control medium or in medium supplemented with CNTF or staurosporine. The cultures were analyzed by immunocytochemistry with the monoclonal antibody Rho4D2, which recognizes chicken rhodopsin and green cone pigment, and by reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis to investigate visual pigment expression at the mRNA level. RESULTS: CNTF increased the number of Rho4D2-immunoreactive photoreceptors in retinal cell cultures, in agreement with previous reports. RT-PCR and Northern blot analysis, however, showed that rhodopsin mRNA was undetectable in both control and CNTF-treated cultures but that CNTF induced significant increases in mRNA levels for the green cone pigment. Staurosporine-treated cultures also had more Rho4D2-immunoreactive cells than control cultures, but this increase was accompanied by induction of rhodopsin expression, with concomitant decreases in levels of green cone pigment mRNA. No significant differences were found between CNTF- or staurosporine-treated cultures and the corresponding control cultures regarding the red cone pigment, which was expressed in all cases, and the blue and violet pigments, which were not detected in any of the samples. CONCLUSIONS: The results suggest that multiple regulatory systems control visual pigment expression during differentiation of chick embryo photoreceptor cells. CNTF appears to stimulate specifically the differentiation of green cones, without the previously suggested effects on the differentiation of rod photoreceptors in ED 8 chick retinal cultures.

Animals↗
PubMed2000

Bipolar cells use kainate and AMPA receptors to filter visual information into separate channels.

Unlike cone photoreceptors, whose light responses have a uniform time course, retinal ganglion cells are tuned to respond to different temporal components in a changing visual scene. The signals in a mammalian cone flow to three to five morphologically distinct "OFF" bipolar cells at a sign-conserving, glutamatergic synapse. By recording simultaneously from pairs of synaptically connected cones and OFF bipolar cells, I now show that each morphological type of OFF bipolar cell receives its signal through a different AMPA or kainate receptor. The characteristic rate at which each receptor recovers from desensitization divides the cone signal into temporal components. Temporal processing begins at the first synapse in the visual system.

Animals↗
PubMedDate not supplied

Probing inner retinal circuits in the rod pathway: a comparison of c-fos activation in mutant mice.

We have used wild-type mice and mice possessing defects in specific retinal circuits in order to more clearly define functional circuits of the inner retina. The retina of the nob mouse lacks communication between photoreceptors and depolarizing bipolar cells (DBCs). Thus, all light driven activity in the nob mouse is mediated via remaining hyperpolarizing bipolar cell (HBC) circuits. Transducin null (Tr alpha-/-) mice lack rod photoreceptor activity and thus remaining retinal circuits are solely generated via cone photoreceptor activity. Activation in inner retinal circuits in each of these mice was identified by monitoring light-induced expression of an immediate early gene, c-fos. The number of cells expressing c-fos in the inner retina was dependent upon stimulus intensity and was altered in a systematic fashion in mice with known retinal mutations. To determine whether c-fos is activated via circuits other than photoreceptors in the outer retina, we examined c-fos expression in tulp1-/- mice that lack photoreceptors in the outer retina; these mice showed virtually no c-fos activity following light exposure. Double-labeling immunohistochemical studies were carried out to more clearly define the population of c-fos expressing amacrine cells. Our results indicate that c-fos may be used to map functional circuits in the retina.

Animals↗
PubMed2001

Rod-cone interactions: developmental and clinical significance.

During the last decade, numerous research reports have considerably improved our knowledge about the physiopathology of retinal degenerations. Three non-mutually exclusive general areas dealing with therapeutic approaches have been proposed; gene therapy, pharmacology and retinal transplantations. The first approach involving correction of the initial mutation, will need a great deal of time and further development before becoming a therapeutic tool in human clinical practice. The observation that cone photoreceptors, even those seemingly unaffected by any described anomaly, die secondarily to rod disappearance related to mutations expressed specifically in the latter, led us to study the interactions between these two photoreceptor populations to search for possible causal links between rod degeneration and cone death. These in vivo and in vitro studies suggest that paracrine interactions between both cell types exist and that rods are necessary for continued cone survival. Since the role of cones in visual perception is essential, pending the identification of the factors mediating these interactions underway, rod replacement by transplantation and/or neuroprotection by trophic factors or alternative pharmacological means appear as promising approaches for limiting secondary cone loss in currently untreatable blinding conditions.

Animals↗
PubMed1999

Role of nitric oxide in photoreceptor survival in embryonic chick retinal cell culture.

The presence of nitric oxide synthase (NOS) in chick retina during development has allowed us to study the role of nitric oxide (NO) during retinal differentiation in dissociated chick retinal cell culture from embryonic day 6. We have demonstrated the presence of nicotinamide adenine dinucleotide phosphate diaphorase staining in these cultures after 3 days in vitro (Div), with a maximal intensity after 8 Div, corresponding to embryonic day 14. Immunohistochemistry studies confirmed the presence of the two isoforms of NOS, NOS-I and -III, in dissociated retinal cell cultures at 8 Div. Addition of NG-monomethyl-L-arginine, a NOS inhibitor, to retinal cell cultures prevented NO production but did not modify the appearance and the survival of ganglion and amacrine cells. However, immunohistochemical analysis with distinct markers for photoreceptor cells (rods and cones) showed that inhibition of endogenous NOS in retinal cell cultures prevented the developmental decrease of rod number between 5 and 8 Div, thus supporting the hypothesis that NO may be involved in the cell death of rods during the development of the retina.

Animals↗
PubMed2007

Dynamic expression of the basic helix-loop-helix transcription factor neuroD in the rod and cone photoreceptor lineages in the retina of the embryonic and larval zebrafish.

NeuroD is a basic helix-loop-helix (bHLH) transcription factor critical for determining neuronal cell fate and regulating withdrawal from the cell cycle. We showed previously that, in goldfish, neuroD is expressed in the rod photoreceptor lineage, and we inferred that neuroD is also expressed in a subset of amacrine cells and nascent cone photoreceptors. Here we extended that study by examining the temporal and spatial expression pattern of neuroD in the embryonic and larval zebrafish and by identifying the cell types that express this gene. NeuroD expression in the developing zebrafish retina is dynamic, spanning early retinogenesis and the maturation of cone photoreceptors. In early retinogenesis neuroD expression expands from a small patch in the ventronasal retina, through the remaining retinal neuroepithelium. As retinogenesis progresses, neuroD expression becomes restricted to amacrine cells, immature cones, and cells of rod and cone lineages. This expression achieves an adult pattern by 96 hours postfertilization (hpf), whereupon the temporal pattern of neuroD expression in central retina is spatially recapitulated at the germinative margin. The cellular pattern of expression suggests that neuroD regulates aspects of rod and cone genesis, but through separate cellular lineages. Furthermore, neuroD is coexpressed with the cone-rod-homeobox transcription factor (Crx) in putative cone progenitors and nascent cone photoreceptors, suggesting that, in the zebrafish retina, as in other vertebrate retinas, similar genetic cascades regulate photoreceptor genesis and maturation.

Animals↗
PubMed2003

Topography of photoreceptors and retinal ganglion cells in the spotted hyena (Crocuta crocuta).

The spatial distributions of photoreceptors and retinal ganglion cells were examined in the spotted hyena (Crocuta crocuta). Two populations of cones were identified by immunocytochemical labeling. The hyena retina contains approximately 2.3 million middle- to long-wavelength sensitive (M/L) cones that reach peak densities of about 7,500/mm(2) in the vicinity of the optic nerve head. A sparser population of short-wavelength sensitive (S) cones, totaling about 0.3 million, was also detected. There is a striking disparity in the spatial distributions of the two cone types with S cones achieving peak density in a region located well below the optic nerve head. The differences in the spatial distributions of the two cone types have implications both for visual sensitivity and for color vision. Hyena rods outnumber cones by about 100:1 with rod density falling off modestly along a central-peripheral gradient. Ganglion cells were identified in retinal wholemounts by Nissl staining patterns. Their distribution defines a prominent visual streak with highest spatial packing (approx. 4,200/mm(2)) in an area centralis that is located in the temporal retina. The total number of ganglion cells is estimated at about 260,000. Using standard assumptions the maximum spatial resolution of the spotted hyena is calculated to be about 8.4 cycles/degree, a value similar to estimates obtained for other terrestrial carnivores.

Animals↗
PubMed2003

Goalpha labels ON bipolar cells in the tiger salamander retina.

By using double-label immunocytochemistry and confocal microscopy, we studied rod and cone synaptic contacts, photoreceptor-bipolar cell convergence, and patterns of axon terminal ramification of ON bipolar cells in the tiger salamander retina. An antibody to recoverin, a calcium-binding protein found in photoreceptors and other retinal neurons in various vertebrates, differentially labeled rods and cones by lightly staining rod cell bodies, axons, and synaptic pedicles and heavily staining cone cell bodies and pedicles. An antibody to G(oalpha) labeled most ON bipolar cells, with axon terminals ramified mainly in strata 6-9 and a minor band in stratum 3 of the inner plexiform layer (IPL). Stratum 10 of the IPL was G(oalpha) negative, and previous studies showed that axon terminals of rod-dominated ON bipolar cells are monostratified in that stratum. The axonal morphology of G(oalpha)-positive cells resembled that of the cone-dominated (DBC(C)) or mixed rod and cone ON (DBC(M)) bipolar cells. The G(oalpha)-positive dendritic processes made close contact with all cone pedicles and superficial contact with some rod pedicles, consistent with the idea that G(oalpha) subunits are present in DBC(C)s and DBC(M)s. The size and density of these cells were analyzed, and their spatial distributions were determined. To our knowledge, this is the first study to characterize photoreceptor inputs and axon terminal morphology of a population of ON bipolar cell with the use of a G(oalpha) antibody as an immunomarker in the salamander retina.

Ambystoma↗