Combining ganglion cell topology and data of patients with glaucoma to determine a structure-function map

PURPOSE

Aligned nanofibers from polypyrrole/graphene as electrodes for regeneration of optic nerve via electrical stimulation

The damage of optic nerve will cause permanent visual field loss and irreversible ocular diseases, such as glaucoma. The damage of optic nerve is mainly derived from the atrophy, apoptosis or death of retinal ganglion cells (RGCs). Though some progress has been achieved on electronic retinal implants that can electrically stimulate undamaged parts of RGCs or retina to transfer signals, stimulated self-repair/regeneration of RGCs has not been realized yet. The key challenge for development of electrically stimulated regeneration of RGCs is the selection of stimulation electrodes with a sufficient safe charge injection limit (Q(inj), i.e., electrochemical capacitance). Most traditional electrodes tend to have low Q(inj) values. Herein, we synthesized polypyrrole functionalized graphene (PPy-G) via a facile but efficient polymerization-enhanced ball milling method for the first time. This technique could not only efficiently introduce electron-acceptor nitrogen to enhance capacitance, but also remain a conductive platform-the π-π conjugated carbon plane for charge transportation. PPy-G based aligned nanofibers were subsequently fabricated for guided growth and electrical stimulation (ES) of RGCs. Significantly enhanced viability, neurite outgrowth and antiaging ability of RGCs were observed after ES, suggesting possibilities for regeneration of optic nerve via ES on the suitable nanoelectrodes.

Effect of vascular endothelial growth factor upregulation on retinal gene expression in the Kimba mouse

Background:  The Kimba mouse carries a human vascular endothelial growth factor transgene causing retinal neovascularisation similar to that seen in diabetic retinopathy. Here, we examine the relationship between differential gene expression induced by vascular endothelial growth factor overexpression and the architectural changes that occur in the retinae of these mice.

Methods:  Retinal gene expression changes in juvenile and adult Kimba mice were assayed by microarray and compared with age-matched wild-type littermates. Transcription of selected genes was validated by quantitative real-time polymerase chain reaction. Protein translation was determined using immunohistochemistry and enzyme-linked immunosorbent assay.

Results:  Semaphorin 3C was upregulated, and nuclear receptor subfamily 2, group 3, member 3 (Nr2e3) was downregulated in juvenile Kimba mice. Betacellulin and endothelin 2 were upregulated in adults. Semaphorin 3C colocalized with glial fibrillary acidic protein in Müller cells of Kimba retinae at greater signal intensities than in wild type. Endothelin 2 colocalised to Müller cell end feet and extended into the outer limiting membrane. Endothelin receptor type B staining was most pronounced in the inner nuclear layer, the region containing Müller cell somata.

Conclusions:  An early spike in vascular endothelial growth factor induced significant long-term retinal neovascularisation associated with changes to the retinal ganglion, photoreceptor and Müller cells. Overexpression of vascular endothelial growth factor led to dysregulation of photoreceptor metabolism through differential expression of Nr2e3, endothelin 2, betacellulin and semaphorin 3C. Alterations in the expression of these genes may therefore play key roles in the pathological mechanisms that result from retinal neovascularisation.

Photoreceptor topography and spectral sensitivity in the common brushtail possum (Trichosurus vulpecula)

Marsupials are believed to be the only non-primate mammals with both trichromatic and dichromatic color vision. The diversity of color vision systems present in marsupials remains mostly unexplored. Marsupials occupy a diverse range of habitats, which may have led to considerable variation in the presence, density, distribution, and spectral sensitivity of retinal photoreceptors. In this study we analyzed the distribution of photoreceptors in the common brushtail possum (Trichosurus vulpecula). Immunohistochemistry in wholemounts revealed three cone subpopulations recognized within two spectrally distinct cone classes. Long-wavelength sensitive (LWS) single cones were the largest cone subgroup (67-86%), and formed a weak horizontal visual streak (peak density 2,106 ± 435/mm2) across the central retina. LWS double cones were strongly concentrated ventrally (569 ± 66/mm2), and created a "negative" visual streak (134 ± 45/mm2) in the central retina. The strong regionalization between LWS cone topographies suggests differing visual functions. Short-wavelength sensitive (SWS) cones were present in much lower densities (3-10%), mostly located ventrally (179 ± 101/mm2). A minority population of cones (0-2.4%) remained unlabeled by both SWS- and LWS-specific antibodies, and may represent another cone population. Microspectrophotometry of LWS cone and rod visual pigments shows peak spectral sensitivities at 544 nm and 500 nm, respectively. Cone to ganglion cell convergences remain low and constant across the retina, thereby maintaining good visual acuity, but poor contrast sensitivity during photopic vision. Given that brushtail possums are so strongly nocturnal, we hypothesize that their acuity is set by the scotopic visual system, and have minimized the number of cones necessary to serve the ganglion cells for photopic vision.

Ultraviolet vision and mate choice in zebra finches

SEXUAL selection is one of the most actively studied areas of evolutionary biology(1-3), and ever since Darwin(1) birds have been probably the most popular taxon for testing the predictions about colour variation. Humans have been used to assess 'colour', an approach which may be flawed(4,5) as many birds see in the ultraviolet (to which humans are blind), and have at least four spectral classes of retinal cone cells (humans have only three), Here we report experiments on zebra finches which test the hypothesis that the ultraviolet waveband (300-400 nm) is used in avian mate-choice decisions. We found that the ultraviolet is used, and that it probably contributes to hue perception. This finding may have,vide implications for future studies of avian sexual selection and colour, and supports one hypothesized function of avian ultraviolet vision, the role of which is largely unknown.(4,6,7)

Differential expression of ATP7A, ATP7B and CTR1 in adult rat dorsal root ganglion tissue

Background: ATP7A, ATP7B and CTR1 are metal transporting proteins that control the cellular disposition of copper and platinum drugs, but their expression in dorsal root ganglion (DRG) tissue and their role in platinum-induced neurotoxicity are unknown. To investigate the DRG expression of ATP7A, ATP7B and CTR1, lumbar DRG and reference tissues were collected for real time quantitative PCR, RT-PCR, immunohistochemistry and Western blot analysis from healthy control adult rats or from animals treated with intraperitoneal oxaliplatin (1.85 mg/kg) or drug vehicle twice weekly for 8 weeks.
Results: In DRG tissue from healthy control animals, ATP7A mRNA was clearly detectable at levels similar to those found in the brain and spinal cord, and intense ATP7A immunoreactivity was localised to the cytoplasm of cell bodies of smaller DRG neurons without staining of satellite cells, nerve fibres or co-localisation with phosphorylated heavy neurofilament subunit (pNF-H). High levels of CTR1 mRNA were detected in all tissues from healthy control animals, and strong CTR1 immunoreactivity was associated with plasma membranes and vesicular cytoplasmic structures of the cell bodies of larger-sized DRG neurons without co-localization with ATP7A. DRG neurons with strong expression of ATP7A or CTR1 had distinct cell body size profiles with minimal overlap between them. Oxaliplatin treatment did not alter the size profile of strongly ATP7A-immunoreactive neurons but significantly reduced the size profile of strongly CTR1-immunoreactive neurons. ATP7B mRNA was barely detectable, and no specific immunoreactivity for ATP7B was found, in DRG tissue from healthy control animals.
Conclusions: In conclusion, adult rat DRG tissue exhibits a specific pattern of expression of copper transporters with distinct subsets of peripheral sensory neurons intensely expressing either ATP7A or CTR1, but not both or ATP7B. The neuron subtype-specific and largely non-overlapping distribution of ATP7A and CTR1 within rat DRG tissue may be required to support the potentially differing cuproenzyme requirements of distinct subsets of sensory neurons, and could influence the transport and neurotoxicity of oxaliplatin.

Retinal tissue thickness is reduced in diabetic peripheral neuropathy

AIM: To investigate the relationship between diabetic peripheral neuropathy (DPN) and retinal tissue thickness.

METHODS: Full retinal thickness in the central retinal, parafoveal, and perifoveal zones and thickness of the ganglion cell complex and retinal nerve fiber layer (RNFL) were assessed in 193 individuals (84 with type 1 diabetes, 67 with type 2 diabetes, and 42 healthy controls) using spectral domain optical coherence tomography. Among those with diabetes, 44 had neuropathy defined using a modified neuropathy disability score recorded on a 0-10 scale. Multiple regression analysis was performed to investigate the relationship between diabetic neuropathy and retinal tissue thickness, adjusted for the presence of diabetic retinopathy (DR), age, sex, duration of diabetes, and HbA1c levels.

RESULTS: In individuals with diabetes, perifoveal thickness was inversely related to the severity of neuropathy (p < 0.05), when adjusted for age, sex, duration of diabetes, and HbA1c levels. DR was associated with reduced thickness in parafovea (p < 0.01). The RNFL was thinner in individuals with greater degrees of neuropathy (p < 0.04).

CONCLUSIONS: DPN is associated with structural compromise involving several retinal layers. This compromise may represent a threat to visual integrity and therefore warrants examination of functional correlates.

Retinal tissue thickness in type 1 and type 2 diabetes

BACKGROUND: The objective was to investigate full retinal and inner retinal thickness in individuals with type 1 and type 2 diabetes. METHODS: Eighty-four individuals with type 1 diabetes (T1DM), 67 individuals with type 2 diabetes (T2DM) and 42 non-diabetic individuals (control group) were enrolled. Participants underwent full retinal thickness evaluation in the central retinal, parafoveal and perifoveal zones and in the retinal nerve fibre layer (RNFL) and ganglion cell complex (GCC), using spectral domain optical coherence tomography. As a preliminary step, the key variables of interest - age, sex, diabetic retinopathy (DR), duration of diabetes and HbA1c levels - were analysed and compared between the three groups. Full retinal thickness, RNFL and GCC thicknesses were also compared between the groups. The relationship between the type of diabetes and retinal tissue thickness was explored, adjusting for the five potential confounders. RESULTS: Compared to individuals with T1DM, individuals with T2DM had significantly reduced full retinal thickness in the parafovea and perifovea and reduced RNFL and GCC thickness. The mean differences were six (p = 0.020), seven (p = 0.008), six (p = 0.021) and four micrometres (p = 0.013) for the parafovea, perifovea, RNFL and GCC thicknesses, respectively. Thicknesses within the central zone (p = 0.018) and at the parafovea (p = 0.007) were significantly reduced in T2DM when compared to the control group. After adjusting for age, sex, diabetic retinopathy, duration of diabetes and HbA1c levels, the relationship between type of diabetes and retinal tissue thickness was not statistically significant (p > 0.056). CONCLUSION: Retinal tissue thickness is not significantly different between type 1 and type 2 diabetes, when adjusted for age, sex, diabetic retinopathy, duration of diabetes and HbA1c levels.

Retinal thickness profile of individuals with diabetes

PURPOSE: To examine the retinal thickness profiles of individuals with and without diabetic retinopathy (DR).

METHODS: Full retinal thickness in the central zone, overall and hemisphere thicknesses of the parafovea and perifovea, ganglion cell complex (GCC) thickness and retinal nerve fibre layer (RNFL) thickness were assessed in 185 individuals using spectral domain optical coherence tomography (88 individuals with diabetes but no DR, 55 with DR, and 42 non-diabetic controls). The DR group comprised of 60% of participants with very mild non-proliferative diabetic retinopathy (NPDR) (representing microaneurysms only) and 40% with mild NPDR (hard exudates, cotton-wool spots, and/or mild retinal haemorrhages). Regression analysis was performed to determine the factors associated with retinal tissue thickness, taking into account, age, sex, presence of DR, duration of diabetes, HbA1c levels and type of diabetes.

RESULTS: The mean (S.D.) of the overall parafoveal thickness was 306 (16) in the DR group and 314 (14) in the control group (p = 0.02). The mean (S.D.) of the superior hemisphere parafoveal thickness was 309 (16) in the DR group and 318 (14) in the control group (p = 0.02). The mean (S.D.) of the inferior hemisphere parafoveal thickness was 303 (17) in the DR group and 311 (15) in the control group (p = 0.02). There were no significant differences in retinal thickness between groups in the central zone (p = 0.27) or perifovea (p > 0.41). Neither the overall nor the hemisphere RNFL (p > 0.75) and GCC thickness (p > 0.37) were significantly different between the groups. Regression analysis revealed that parafoveal thickness in diabetic individuals was reduced in association with presence of DR (B = -5.9 μm, p = 0.02) and with advancing age (B = -4.5 μm, p = 0.004, for every 10 year increase in age) when adjusted for sex, duration of diabetes, HbA1c levels and type of diabetes.

CONCLUSION: The inner macula is thinner in the presence of clinical signs of diabetic retinopathy and is compounded by advancing age. The influence of any macular oedema or that by cotton-wool spots could not be ruled out and may still confound these results.