Inner Retinal Layers Research Articles

Possible implications of acid-sensing ion channels in ischemia-induced retinal injury in rats

Retinal ischemia in eyes with diabetic retinopathy and retinal vein occlusion leads to local tissue acidosis. Acid-sensing ion channels (ASICs) are expressed in photoreceptors and other neurons in the retina, and may play a role in acid-induced cell injury. The purpose of this study was to investigate the neuroprotective effects of amiloride, an ASIC blocker, on induced retinal ischemia in rats. Transient retinal ischemia was induced in male Long-Evans rats by the temporary ligation of the optic nerve. Just before the induction of ischemia, the experimental eyes underwent intravitreal injection of amiloride. On day 7, the retinal damage in eyes that underwent amiloride treatment (and in those that did not undergo the treatment) was evaluated by histology and electroretinogram (ERG). Transient retinal ischemia caused retinal degeneration with thinning of the inner layer of the retina. The blockage of ASICs with amiloride significantly prevented retinal degeneration. ERG demonstrated that the reduction in a- and b-wave amplitudes induced by the transient retinal ischemia was significantly prevented by the application of amiloride. The present study suggests that ASICs might, at least in part, play a pathophysiological role in ischemia-induced neurodegeneration. Blockage of ASICs may have a potential neuroprotective effect in ocular ischemic diseases.

Effect of brain-derived neurotrophic factor on c-jun expression in the rd mouse retina.

To determine the location of c-jun protein, dynamic changes in c-jun mRNA and protein expression, and ultrastructure characteristics in the rd mouse retina, following a single dose of brain-derived neurotrophic factor (BDNF) in a short period of time. A single intravitreal injection of BDNF at two dosages (25µg/L or 50µg/L) was given to the right eye of the rd mouse at age 2 and 3 weeks respectively. Two weeks after injection, the location of c-jun protein in the retina was observed by immunofluorescence detection, c-jun mRNA and protein expression in retinas were detected by quantitative real time polymerase chain reaction (RT-PCR) and western immunoblotting analysis, ultrastructure characteristics of retinas were detected by transmission electron microscope (TEM) observation. c-jun protein was expressed in the inner nuclear layer (INL) of retina. BDNF at two dosages (25µg/L and 50µg/L) increased c-jun mRNA expression at PN-4 weeks respectively (P(1)=0.019, P(2)=0.021). 50µg/L BDNF increased c-jun protein expression at PN-4 weeks (P =0.000). The retinal ultrastructure was improved. The effects of BDNF exerts on the c-jun expression in the retina are dose-dependent and time-dependent, which may mediate photoreceptor rescue indirectly in the pathological process of retinitis pigmentosa (RP) at early stage.

Expression of prolyl hydroxylase-2 in retina of diabetic rats

Objective To investigate the expression and prolyl hydroxylase (PHD)-2 in retina of diabetic rats.Methods Wistar rats were randomly divided into the control group (n=48) and the diabetes group (n=60).The rats in diabetes group were induced with streptozotocin (STZ) injection creating a diabetic retinopathy model.The same volume of citric acid buffer was injected into the rats in the control group.Fluorescence microscope was used to observe the retinal vaseulature at one,three and six months after injection.Evans blue perfusion was used to detect the blood-retinal barrier (BRB) permeability.Immunohistochemical staining was used to observe the distribution of PHD-2 positive staining.Western blot was used to measure the protein expression of PHD-2,hypoxia inducible factor(HIF)-1α and vascular endothelial growth factor (VEGF) every month from one to six months after injection.Results The vascularization was normal and form was clear in retina of rats in control group.The retinal blood vessels of rats in diabetes group showed significantly increased fluorescence.Compared with the control group,the BRB permeability was significantly increased in diabetes group (P＜0.05).Abundant expression of PHD-2protein was detected in the inner layers of retina in control and diabetes group.Compared with the control group,the PHD-2 expression was decreased in diabetes group at one and two months after iujection (t=16.230,16.390;P＜0.05).The HIF-1α expression was significantly increased in diabetes group at one,two and three months after injection (t=27.073,36.709,10.176; P＜0.05).The VEGF expression was significantly increased in diabetes group every month from one to six months after injection (t =13.547,31.984,21.897,8.912,9.019,14.046; P＜0.05).Conclusions There is abundant expression of PHD-2 in the inner layer of retina in diabetic rats.PHD-2 may play an important role in diabetic retinopathy,which is correlated with VEGF. Key words: Diabetic retinopathy/pathophysiology; Dioxygenases/physiology; Animal experimentation

The Expression of irx7 in the Inner Nuclear Layer of Zebrafish Retina Is Essential for a Proper Retinal Development and Lamination

Irx7, a member in the zebrafish iroquois transcription factor (TF) family, has been shown to control brain patterning. During retinal development, irx7's expression was found to appear exclusively in the inner nuclear layer (INL) as soon as the prospective INL cells withdraw from the cell cycle and during retinal lamination. In Irx7-deficient retinas, the formation of a proper retinal lamination was disrupted and the differentiation of INL cell types, including amacrine, horizontal, bipolar and Muller cells, was compromised. Despite irx7's exclusive expression in the INL, photoreceptors differentiation was also compromised in Irx7-deficient retinas. Compared with other retinal cell types, ganglion cells differentiated relatively well in these retinas, except for their dendritic projections into the inner plexiform layer (IPL). In fact, the neuronal projections of amacrine and bipolar cells into the IPL were also diminished. These indicate that the retinal lamination issue in the Irx7-deficient retinas is likely caused by the attenuation of the neurite outgrowth. Since the expression of known TFs that can specify specific retinal cell type was also altered in Irx7-deficient retinas, thus the irx7 gene network is possibly a novel regulatory circuit for retinal development and lamination.

GABAA Receptors Containing the α2 Subunit Are Critical for Direction-Selective Inhibition in the Retina

Far from being a simple sensor, the retina actively participates in processing visual signals. One of the best understood aspects of this processing is the detection of motion direction. Direction-selective (DS) retinal circuits include several subtypes of ganglion cells (GCs) and inhibitory interneurons, such as starburst amacrine cells (SACs). Recent studies demonstrated a surprising complexity in the arrangement of synapses in the DS circuit, i.e. between SACs and DS ganglion cells. Thus, to fully understand retinal DS mechanisms, detailed knowledge of all synaptic elements involved, particularly the nature and localization of neurotransmitter receptors, is needed. Since inhibition from SACs onto DSGCs is crucial for generating retinal direction selectivity, we investigate here the nature of the GABA receptors mediating this interaction. We found that in the inner plexiform layer (IPL) of mouse and rabbit retina, GABAA receptor subunit α2 (GABAAR α2) aggregated in synaptic clusters along two bands overlapping the dendritic plexuses of both ON and OFF SACs. On distal dendrites of individually labeled SACs in rabbit, GABAAR α2 was aligned with the majority of varicosities, the cell's output structures, and found postsynaptically on DSGC dendrites, both in the ON and OFF portion of the IPL. In GABAAR α2 knock-out (KO) mice, light responses of retinal GCs recorded with two-photon calcium imaging revealed a significant impairment of DS responses compared to their wild-type littermates. We observed a dramatic drop in the proportion of cells exhibiting DS phenotype in both the ON and ON-OFF populations, which strongly supports our anatomical findings that α2-containing GABAARs are critical for mediating retinal DS inhibition. Our study reveals for the first time, to the best of our knowledge, the precise functional localization of a specific receptor subunit in the retinal DS circuit.

Recent progress in molecular genetics and therapy for X-linked juvenile retinoschisis

Juvenile retinoschisis (RS or XLRS,MIM#312700)is a rare X-linked inherited disorder,mainly affects bilateral retina,and is characterized by cartwheel-like changes of the macular region of the retina and schisis or splitting within the inner retinal layers,leading to visual deterioration.The electroretinogram is beneficial in the diagnosis of juvenile retinoschisis.The a-wave can be of normal or nearly normal amplitude in this disorder,whereas the amplitude of the b-wave is appreciably reduced,giving a decrease in the proportion of b/a.The responsible gene,XLRSl,maps to Xp22 and was identified by positional cloning.This paper makes a brief review about the latest XLRS research of pathogenesis,animal experiments,clinical therapy,and 25 references are cited. Key words: X-linked juvenile retinoschisis; Therapeutics; Model

3D segmentation of SBFSEM images of neuropil by a graphical model over supervoxel boundaries

The segmentation of large volume images of neuropil acquired by serial sectioning electron microscopy is an important step toward the 3D reconstruction of neural circuits. The only cue provided by the data at hand is boundaries between otherwise indistinguishable objects. This indistinguishability, combined with the boundaries becoming very thin or faint in places, makes the large body of work on region-based segmentation methods inapplicable. On the other hand, boundary-based methods that exploit purely local evidence do not reach the extremely high accuracy required by the application domain that cannot tolerate the global topological errors arising from false local decisions. As a consequence, we propose a supervoxel merging method that arrives at its decisions in a non-local fashion, by posing and approximately solving a joint combinatorial optimization problem over all faces between supervoxels. The use of supervoxels allows the extraction of expressive geometric features. These are used by the higher-order potentials in a graphical model that assimilate knowledge about the geometry of neural surfaces by automated training on a gold standard. The scope of this improvement is demonstrated on the benchmark dataset E1088 (Helmstaedter et al., 2011) of 7.5billionvoxels from the inner plexiform layer of rabbit retina. We provide C++ source code for annotation, geometry extraction, training and inference.

Inhibition of hypoxia-induced [ 3H]glycine release from chicken retina by the glycine transporter type-1 (GlyT-1) inhibitors NFPS and Org-24461

Chicken posterior eyecup lined by the retina were prepared, loaded with [ 3H]glycine and superfused in order to determine its release in various experimental conditions. Electrical field stimulation of the retina evoked [ 3H]glycine release with a voltage- and frequency-dependent manner and this release may be originated from glycinergic amacrine cell processes of the inner plexiform layer of the retina. Glycine released from an abundance of different amacrine cells may modulate retinal circuitry by activation of inhibitory glycine receptors and by acting as a coagonist on N-methyl- d-aspartate receptors on AII amacrine cells and retinal ganglion cells. The latter effect of glycine may be modulated by glycine transporter type-1. Cells with glycine transporter type-1 immunopositive staining were visualized in the inner nuclear layer and dens immunolabeling was also detected throughout the inner plexiform layer of chicken retina. Glycine and the substrate-type glycine transporter type-1 inhibitor sarcosine increased [ 3H]glycine release from glycinergic amacrine cells and/or glial cells by extrusion of glycine from cytoplasmic pools by homo- and heteroexchange mechanisms. Deprivation of oxygen and glucose from the buffer used for superfusion evoked a marked increase in [ 3H]glycine efflux, an effect probably due to reverse mode operation of glycine transporter type-1. The non-transportable glycine transporter type-1 inhibitors NFPS and Org-24461, which did not alter [ 3H]glycine efflux from isolated chicken retina by themselves in normoxic condition, inhibited oxygen and glucose deprivation-induced [ 3H]glycine release. It is concluded that reduction of the N-methyl- d-aspartate receptor coagonist glycine concentrations in hypoxic conditions by glycine transporter type-1 inhibitors may decrease N-methyl- d-aspartate receptor-mediated neuronal toxicity and cell death in retinal tissue.

Dynamics of Human Foveal Development after Premature Birth

To determine the dynamic morphologic development of the human fovea in vivo using portable spectral domain-optical coherence tomography (SD-OCT). Prospective, observational case series. Thirty-one prematurely born neonates, 9 children, and 9 adults. Sixty-two neonates were enrolled in this study. After examination for retinopathy of prematurity (ROP), SD-OCT imaging was performed at the bedside in nonsedated infants aged 31 to 41 weeks postmenstrual age (PMA) (= gestational age in weeks + chronologic age) and at outpatient follow-up ophthalmic examinations. Thirty-one neonates met eligibility criteria. Nine children and nine adults without ocular pathology served as control groups. Semiautomatic retinal layer segmentation was performed. Central foveal thickness, foveal to parafoveal (FP) ratio (central foveal thickness divided by thickness 1000 μm from the foveal center), and 3-dimensional thickness maps were analyzed. In vivo determination of foveal morphology, layer segmentation, analysis of subcellular changes, and spatiotemporal layer shifting. In contrast with the adult fovea, several signs of immaturity were observed in the neonates: a shallow foveal pit, persistence of inner retinal layers (IRLs), and a thin photoreceptor layer (PRL) that was thinnest at the foveal center. Three-dimensional mapping showed displacement of retinal layers out of the foveal center as the fovea matured and the progressive formation of the inner/outer segment band in the opposite direction. The FP-IRL ratios decreased as IRL migrated before term and minimally after that, whereas FP-PRL ratios increased as PRL subcellular elements formed closer to term and into childhood. A surprising finding was the presence of cystoid macular edema in 58% of premature neonates that appeared to affect inner foveal maturation. This study provides the first view into the development of living cellular layers of the human retina and of subcellular specialization at the fovea in premature infant eyes using portable SD-OCT. Our work establishes a framework of the timeline of human foveal development, allowing us to identify unexpected retinal abnormalities that may provide new keys to disease activity and a method for mapping foveal structures from infancy to adulthood that may be integral in future studies of vision and visual cortex development. Proprietary or commercial disclosure may be found after the references.

Surgical macular decompression for macular edema in retinal vein occlusion.

Recently, a number of surgical and laser approaches have been used to improve visual outcome in cases of central retinal vein occlusion (CRVO) and branch retinal vein occlusion (BRVO). Intravitreal steroid injection alone appears to offer only temporary improvement at best. Radial optic neurotomy for CRVO and arteriovenous adventitial sheathotomy for BRVO are the most frequently utilized surgical procedures for these conditions, but evidence regarding efficacy is still lacking. We have suggested that macular decompression by internal limiting membrane (ILM) peeling may reduce macular edema and hemorrhage and improve visual acuity by relieving elevated intraretinal tissue pressure and facilitating egress of blood and extracellular fluid out of inner retinal layers into the vitrectomized vitreous cavity. 50 cases of severe visual loss due to macular edema caused by CRVO or BRVO, not eligible for laser photocoagulation, underwent pars plana vitrectomy with removal of preretinal hyaloid, peeling of the ILM stained with indocyanine green dye, air-fluid exchange, and postoperative prone positioning. In all cases, intraretinal blood and retinal thickening diminished within 6 weeks of surgery. Visual acuity improved in 87% of CRVO cases and 68% of BRVO cases. Vision improved and stabilized at 39 days after surgery. Average improvement was 2.6 lines with a 6-line improvement in 1 case. There was no difference in outcome between cases with ischemic or nonischemic features on fluorescein angiography. Macular decompression using vitrectomy and ILM peeling is effective in the treatment of severe visual loss due to macular edema in CRVO and in those BRVO cases that do not qualify for laser photocoagulation.

Two types of tyrosine hydroxylase-immunoreactive neurons in the zebrafish retina

The purpose of the present study is to identify the dopaminergic amacrine (DA) cells in the inner nuclear layer (INL) of zebrafish retina through immunocytochemistry and quantitative analysis. Two types of tyrosine hydroxylase-immunoreactive (TH-IR) cells appeared on the basis of dendritic morphology and stratification patterns in the inner plexiform layer (IPL). The first (DA1) was bistratified, with branching planes in both s1 and s5 of the IPL. The second (DA2) was diffuse, with dendritic processes branched throughout the IPL. DA1 and DA2 cells corresponded morphologically to A on −s1/s5 and A diffuse −1 ( Connaughton et al., 2004). The average number of total TH-IR cells was 1088 ± 79 cells per retina ( n = 5), and the mean density was 250 ± 27 cells/mm 2. Their density was highest in the mid central region of ventrotemporal retina and lowest in the periphery of dorsonasal retina. Quantitatively, 45.71% of the TH-IR cells were DA1 cells, while 54.29% were DA2 cells. No TH-IR cells expressed calbindin D28K, calretinin or parvalbumin, markers for the various INL cells present in several animals. Therefore the TH-IR cells in zebrafish are limited to very specific subpopulations of the amacrine cells.

Identification of two arylalkylamine N-acetyltranferase 1 genes with different developmental expression profiles in the flatfish Solea senegalensis

The existence of two arylalkylamine N-acetyltransferase 1 (Aanat1) genes in the genome of some teleosts has been reported recently by in silico analysis. However, there are no data concerning the similarities and/or differences between them and many questions remain to be answered, such as their expression sites, development, or kinetics. Here, we report the cloning of Aanat1a and Aanat1b cDNAs from the sole retina and show for the first time that at least three Aanat genes are expressed in a vertebrate species. Because melatonin is involved in fish ontogeny, we analyzed the developmental transcript levels of Aanat1a and Aanat1b by quantitative real-time PCR, showing their inverse and stage-specific expression patterns. Aanat1a was more abundant during early than late larval stages. Before metamorphosis, nocturnal expression was higher. At metamorphosis, Aanat1a expression decreased and lost these day-night variations. In contrast, the abundance of Aanat1b transcripts, low during early developing stages, rose significantly throughout metamorphosis. This situation seemed to apply to the adult because Aanat1a expression was lower than Aanat1b expression in the retina of adults, where the former did not exhibit day-night variations, while the latter did so with much higher nocturnal transcript levels. In situ hybridization analysis detected Aanat1a and Aanat1b messengers in the outer and inner nuclear layers of retina. The differences in abundance and distinct day-night expression patterns between Aanat1a and Aanat1b during sole development suggest different functions for these two enzymes as well as the existence of interactions between the melatoninergic and thyroid hormone systems during flatfish metamorphosis.

Three-Dimensional Imaging of Macular Inner Structures in Glaucoma by Using Spectral-Domain Optical Coherence Tomography

To profile macular thickness changes in glaucoma by using three-dimensional spectral-domain optical coherence tomography (3D-SD-OCT). The study included 30 eyes with suspected glaucoma and preperimetric glaucoma (SGPPG) and 35 healthy eyes. The macular thickness, including those of the total retina, nerve fiber layer (NFL), and combined inner retinal layers (IRLs)-NFL+ganglion cell layer (GCL)+inner plexiform layer (IPL)-was measured by 3D-SD-OCT raster scans in a 6 mm(2) region. The average and sectoral thicknesses were calculated on an Early Treatment of Diabetic Retinopathy Study (ETDRS) chart and a ETDRS chart with a 45° rotation (glaucoma sector chart, GSC). The mean IRL thickness was significantly less in the SGPPG eyes than in the healthy eyes, but the mean total retinal and macular NFL thicknesses were not. In the SGPPG eyes, the IRLs were thinner in the outer macula than in the inner macula, in the inferior hemisphere than in the superior hemisphere, and in the temporal hemisphere than in the nasal hemisphere. The significantly thinned sectors were nearly identical on the GSC but only slightly overlapped on the ETDRS chart. The IRLs in the inferior temporal outer sector (GSC) had the greatest area under the receiver operating characteristic curve, which was significantly greater than those for the IRLs over the entire macula, inferior hemiretinal region, and inferior outer hemicircular region (macular subfields), and that for the circumpapillary NFL in the inferior sectors (P = 0.001-0.036). Macular IRL thickness measured by using 3D-SD-OCT is useful for profiling macular atrophy in SGPPG.

Heterogeneity of Glia in the Retina and Optic Nerve of Birds and Mammals

We have recently described a novel type of glial cell that is scattered across the inner layers of the avian retina [1]. These cells are stimulated by insulin-like growth factor 1 (IGF1) to proliferate, migrate distally into the retina, and up-regulate the nestin-related intermediate filament transitin. These changes in glial activity correspond with increased susceptibility of neurons to excitotoxic damage. This novel cell-type has been termed the Non-astrocytic Inner Retinal Glia-like (NIRG) cells. The purpose of the study was to investigate whether the retinas of non-avian species contain cells that resemble NIRG cells. We assayed for NIRG cells by probing for the expression of Sox2, Sox9, Nkx2.2, vimentin and nestin. NIRG cells were distinguished from astrocytes by a lack of expression for Glial Fibrilliary Acidic Protein (GFAP). We examined the retinas of adult mice, guinea pigs, dogs and monkeys (Macaca fasicularis). In the mouse retina and optic nerve head, we identified numerous astrocytes that expressed GFAP, S100β, Sox2 and Sox9; however, we found no evidence for NIRG-like cells that were positive for Nkx2.2, nestin, and negative for GFAP. In the guinea pig retina, we did not find astrocytes or NIRG cells in the retina, whereas we identified astrocytes in the optic nerve. In the eyes of dogs and monkeys, we found astrocytes and NIRG-like cells scattered across inner layers of the retina and within the optic nerve. We conclude that NIRG-like cells are present in the retinas of canines and non-human primates, whereas the retinas of mice and guinea pigs do not contain NIRG cells.

Detection of retinal changes in Parkinson's disease with spectral-domain optical coherence tomography

PurposeThis pilot study investigated whether high-resolution spectral-domain optical coherence tomography (SD-OCT) could detect differences in inner retinal layer (IRL), peripapillary retinal nerve fiber layer (RNFL), and macular thickness between patients with Parkinson’s disease (PD) and controls.MethodsBoth eyes of patients with PD and age-matched controls were imaged with the Heidelberg Spectralis® HRA + OCT. RNFL, IRL, and macular thickness were measured for each eye using Heidelberg software. These measurements were compared with validated, published normal values for macular and RNFL thickness, and compared with matched controls for IRL thickness.ResultsEighteen eyes from nine subjects with PD and 19 eyes of 16 control subjects were evaluated using SD-OCT. The average age of PD patients was 64 years with a range of 52–75 years. The average age of controls was 67 years with a range of 50–81 years. No significant reduction in IRL thickness was detected between PD patients and age-matched controls at 13 points along a 6 mm horizontal section through the fovea. No significant difference in RNFL thickness was detected between PD patients and published normal values. Overall average RNFL thickness was 97 μm for PD patients, which exactly matched the normative database value. However, significant differences in macular thickness were detected in three of nine subfields between PD subjects and published normal values. In PD subjects, the outer superior subfield was 2.8% thinner (P = 0.026), while the outer nasal and inner inferior subfields were 2.8% (P = 0.016) and 2.7% (P = 0.001) thicker compared to published normal values.ConclusionIn this pilot study, significant differences in macular thickness were detected in three of nine subfields by SD-OCT. However, SD-OCT did not detect significant reductions in peripapillary RNFL and IRL thickness between PD patients and controls. This suggests that macular thickness measurements by SD-OCT may potentially be used as an objective, noninvasive, and easily quantifiable in vivo biomarker in PD. Larger, longitudinal studies are needed to explore these relationships further.

Synaptic connections of calbindin-immunoreactive cone bipolar cells in the inner plexiform layer of rabbit retina

In the mammalian retina, information concerning various aspects of an image is transferred in parallel, and cone bipolar cells are thought to play a major role in this parallel processing. We have examined the synaptic connections of calbindin-immunoreactive (IR) ON cone bipolar cells in the inner plexiform layer (IPL) of rabbit retina and have compared these synaptic connections with those that we have previously described for neurokinin 1 (NK1) receptor-IR cone bipolar cells. A total of 325 synapses made by calbindin-IR bipolar axon terminals have been identified in sublamina b of the IPL. The axons of calbindin-IR bipolar cells receive synaptic inputs from amacrine cells through conventional synapses and are coupled to putative AII amacrine cells via gap junctions. The major output from calbindin-IR bipolar cells is to amacrine cell processes. These data resemble our findings for NK1 receptor-IR bipolar cells. However, the incidences of output synapses to ganglion cell dendrites of calbindin-IR bipolar cells are higher compared with the NK1-receptor-IR bipolar cells. On the basis of stratification level and synaptic connections, calbindin-IR ON cone bipolar cells might thus play an important role in the processing of various visual aspects, such as contrast, orientation, and approach sensing, and in transferring rod signals to the ON cone pathway.

Nodavirus associated with pathological changes in adult spotted coralgroupers ( Plectropomus maculatus) in Thailand with viral nervous necrosis

The present study characterized viral nervous necrosis in sea cage-reared adult spotted coralgroupers ( Plectropomus maculatus). Histopathological study showed extensive vacuolation and neuronal necrosis of the olfactory bulb and the optic lobe of the forebrain and the inner and outer nuclear layer of retina. Mild necrosis was observed in the spinal cord. Homogeneous intranuclear inclusion bodies were noted in the hyperplastic and hypertrophic glandular epithelial cells of the swim bladder suggesting viral etiology. Etiological diagnosis of VNN was confirmed by RT-PCR, immunohistochemistry and in situ hybridization. The immunohistochemistry and in situ hybridization gave strongly positive staining in the same area of the infected cells of the brain, spinal cord and retina correlating with histopathological changes. No positive reaction was detectable in the affected gas glandular epithelium and other organs, confirming the consistent neurotropism of this nodavirus. Nodavirus was mainly detected in the olfactory bulb of the brain. The result suggests nasal transmission was the major route of infection.

Eye-specific expression of an ancestral jellyfish PaxB gene interferes with Pax6 function despite its conserved Pax6/Pax2 characteristics

Pax transcription factors are evolutionarily conserved regulators of eye development and can be distinguished on the basis of three functional domains: two DNA-binding domains (the paired domain and the paired-type homeodomain), and the octapeptide motif. PaxB of the eyed cubozoan jellyfish, Tripedalia cystophora, is characterized by a Pax2-like paired domain and octapeptide, and a Pax6-like homeodomain. In mice, functionally distinct Pax6 and Pax2 proteins have unique as well as redundant roles in eye morphogenesis. Here, we show that expression of the jellyfish PaxB gene in mouse embryonic eye tissues impairs normal development of lens and retina. Our data show that PaxB misexpression leads to a downregulation of endogenous Pax6 protein in the prospective lens and in subsets of cells within the inner nuclear layer of transgenic retina. In addition to Pax6 downregulation, the expression of PaxB leads to an almost complete loss of amacrine cells in the adult transgenic retina, a phenotype that differs from a loss-of-function of the Pax6 gene. The present data suggest that PaxB, due to its Pax2-like paired domain and Pax-6 like homeodomain, disturbs the transcriptional network regulated by Pax6 in the developing lens and retina. Taken together, our data suggest that molecular properties of individual mouse Pax2 and Pax6 proteins are essential determinants of mouse eye development and cannot be substituted for by jellyfish PaxB which possesses elements of vertebrate Pax2 and Pax6.

Parvalbumin-immunoreactive neurons in the inner nuclear layer of zebrafish retina

The purpose of this investigation is to characterize parvalbumin-immunoreactive (IR) neurons in the inner nuclear layer (INL) of zebrafish retina through immunocytochemistry, quantitative analysis, and confocal microscopy. In the INL, parvalbumin-IR neurons were located in the inner marginal portion of the INL. On the basis of dendritic stratification in the inner plexiform layer (IPL), at least two types of amacrine cells were IR for parvalbumin. The first one formed distinctive laminar tiers within s4 (PVs4) of the IPL, and the second within s5 (PVs5). The average number of PVs4 cells was 8263 cells per retina (n=3), and the mean density was 1671cells/mm2. The average number of PVs5 cells was 1037 cells per retina (n=3), and the mean density was 210cells/mm2. Quantitatively, 88.9% of anti-parvalbumin labeled neurons were PVs4 cells and 11.1% were PVs5 cells. Their density was highest in the midcentral region of the ventrotemporal retina and lowest in the periphery of the dorsonasal retina. The average regularity index of the PVs4 cell mosaic was 4.09, while the average regularity index of the PVs5 cell mosaic was 3.46. No parvalbumin-IR cells expressed calretinin or disabled-1, markers for AII amacrine cells, in several animals. These results indicate that parvalbumin-IR neurons in zebrafish are limited to specific subpopulations of amacrine cells and the expressional pattern of parvalbumin may not correspond to AII amacrine cells in several other animals. Their distribution suggests that parvalbumin-IR neurons are mainly involved in ON pathway information flow.

The Expression Pattern of Heterogeneous Nuclear Ribonucleoprotein R in Rat Retina

The heterogeneous nuclear ribonucleoproteins (hnRNPs) play important roles in DNA repairing, cell signaling, telomere biogenesis, and in regulating gene expression at both transcriptional and translational levels. In the present study, we demonstrated that the expression of hnRNP-R1 and hnRNP-R2 is developmentally regulated in rat retina. The neural specific isoform hnRNP-R2 is expressed in 7-day postnatal rat retina, but not in the adult retina. The positive immunohistochemistry signal of hnRNP-R1 is extensively distributed in the outer plexiform layer, inner nuclear layer and ganglion cell layer of rat retina. Double staining experiments showed that the positive signal of hnRNP-R1 is distributed in ON-type bipolar cells and localized in the cytoplasm, dendrites and axon terminals. In addition, the hnRNP-R1 distribution is regulated in rat retina during circadian. The present investigation suggests that hnRNP-R may play roles in retinal development and light-elicited cellular activities.