scholarly journals A Battery of Cell- and Structure-specific Markers for the Adult Porcine Retina

2010 ◽  
Vol 58 (4) ◽  
pp. 377-389 ◽  
Author(s):  
Ulrica Englund Johansson ◽  
Sajedeh Eftekhari ◽  
Karin Warfvinge
Keyword(s):  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Experimental Studies ◽  
Müller Cells ◽  
Amacrine Cells ◽  
Specific Cell ◽  
Muller Cells ◽  
Primate Model ◽  
Specific Staining ◽  
Displaced Amacrine Cells

The pig is becoming an increasingly used non-primate model in experimental studies of human retinal diseases and disorders. The anatomy, size, and vasculature of the porcine eye and retina closely resemble their human counterparts, which allows for application of standard instrumentation and diagnostics used in the clinic. Despite many reports that demonstrate immunohistochemistry as a useful method for exploring neuropathologi-cal changes in the mammalian central nervous system, including the pig, the porcine retina has been sparsely described. Hence, to facilitate further immunohistochemical analysis of the porcine retina, we report on the successful use of a battery of antibodies for staining of paraformaldehyde-fixed cryosectioned retina. The following antibodies were evaluated for neuronal cells and structures: recoverin (cones and rods), Rho4D2 (rods), transducin-γ (cones), ROM-1 (photoreceptor outer segments), calbindin (horizontal cells), PKC-α (bipolar cells), parvalbumin (amacrine and displaced amacrine cells), and NeuN (ganglion cells and displaced amacrines). For detecting synaptic connections in fiber layers, we used an antibody against synaptobrevin. For detecting retinal pigment epithelium, we studied antibodies against cytokeratin and RPE65, respectively. The glial cell markers used were bFGF (Müller cells and displaced amacrine cells), GFAP (Müller cells and astrocytes), and vimentin (Müller cells). Each staining effect was evaluated with regard to its specificity, sensitivity, and repro-ducibility in the identification of individual cells, specific cell structures, and fiber layers, respectively. The markers parvalbumin and ROM-1 were tested here for the first time for the porcine retina. All antibodies tested resulted in specific staining of high quality. In conclusion, all immunohistochemical protocols presented here will be applicable in fixed, cryosectioned pig retina.

scholarly journals Muller cells of chicken retina synthesize 11-cis-retinol

1992 ◽  
Vol 285 (3) ◽  
pp. 907-913 ◽  
Author(s):  
S R Das ◽  
N Bhardwaj ◽  
H Kjeldbye ◽  
P Gouras
Keyword(s):  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Light And Electron Microscopy ◽  
Müller Cells ◽  
Neural Retina ◽  
Retinyl Palmitate ◽  
Immunocytochemical Localization ◽  
Muller Cells ◽  
Trans Isomers ◽  
Chicken Retina

The amounts of endogenous retinyl palmitate, retinol and retinaldehyde were measured in the neural retina and retinal pigment epithelium (RPE) of predominantly cone (chicken), rod (rat) and more mixed (cat, human) retinae. The ratio of 11-cis to all-trans isomers of retinyl palmitate and retinol in the neural retina and the RPE increases progressively with the increase in diurnality of the species from rat to chicken. The membrane fractions of both chicken and bovine RPE enzymically isomerize all-trans retinol to 11-cis-retinol. Chicken neural retina membranes enzymically form 11-cis-retinol and all-trans-retinyl palmitate from all-trans-retinol. Light and electron microscopy revealed no contamination of chicken neural retina by RPE. Muller cells from chicken retina were isolated, cultured and characterized by immunocytochemical localization of cellular retinaldehyde-binding protein. Cultured chicken Muller cells form all-trans-retinyl palmitate, 11-cis-retinol and 11-cis-retinyl palmitate from all-trans-retinol and release most of the 11-cis-retinol into the medium. The results indicate that chicken neural retina and Muller cells in particular synthesize 11-cis-retinoids from all-trans-retinol.

Expression of monocarboxylate transporters in rat ocular tissues

2005 ◽  
Vol 288 (2) ◽  
pp. C416-C428 ◽  
Author(s):  
Glyn Chidlow ◽  
John P. M. Wood ◽  
Mark Graham ◽  
Neville N. Osborne
Keyword(s):  
Ciliary Body ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Müller Cells ◽  
Basal Membrane ◽  
Monocarboxylate Transporter ◽  
Muller Cells ◽  
Ciliary Processes ◽  
Unique Distribution ◽  
Plexiform Layers

The aim of the present study was to determine the distribution of monocarboxylate transporter (MCT) subtypes 1-4 in the various structures of the rat eye by using a combination of conventional and real-time RT-PCR, immunoblotting, and immunohistochemistry. Retinal samples expressed mRNAs encoding all four MCTs. MCT1 immunoreactivity was observed in photoreceptor inner segments, Müller cells, retinal capillaries, and the two plexiform layers. MCT2 labeling was concentrated in the inner and outer plexiform layers. MCT4 immunolabeling was present only in the inner retina, particularly in putative Müller cells, and the plexiform layers. No MCT3 labeling could be observed. The retinal pigment epithelium (RPE)/choroid expressed high levels of MCT1 and MCT3 mRNAs but lower levels of MCT2 and MCT4 mRNAs. MCT1 was localized to the apical and MCT3 to the basal membrane of the RPE, whereas MCT2 staining was faint. Although MCT1-MCT4 mRNAs were all detectable in iris and ciliary body samples, only MCT1 and MCT2 proteins were expressed. These were present in the iris epithelium and the nonpigmented epithelium of the ciliary processes. MCT4 was localized to the smooth muscle lining of large vessels in the iris-ciliary body and choroid. In the cornea, MCT1 and MCT2 mRNAs and proteins were detectable in the epithelium and endothelium, whereas evidence was found for the presence of MCT4 and, to a lesser extent, MCT1 in the lens epithelium. The unique distribution of MCT subtypes in the eye is indicative of the pivotal role that these transporters play in the maintenance of ocular function.

scholarly journals Creatine transporter localization in developing and adult retina: importance of creatine to retinal function

2005 ◽  
Vol 289 (4) ◽  
pp. C1015-C1023 ◽  
Author(s):  
Monica L. Acosta ◽  
Michael Kalloniatis ◽  
David L. Christie
Keyword(s):  
Ganglion Cells ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Müller Cells ◽  
Nerve Fibers ◽  
Retinal Function ◽  
Mouse Retina ◽  
Muller Cells ◽  
Creatine Transporter ◽  
Subsequent Decrease

Creatine and phosphocreatine are required to maintain ATP needed for normal retinal function and development. The aim of the present study was to determine the distribution of the creatine transporter (CRT) to gain insight to how creatine is transported into the retina. An affinity-purified antibody raised against the CRT was applied to adult vertebrate retinas and to mouse retina during development. Confocal microscopy was used to identify the localization pattern as well as co-localization patterns with a range of retinal neurochemical markers. Strong labeling of the CRT was seen in the photoreceptor inner segments in all species studied and labeling of a variety of inner neuronal cells (amacrine, bipolar, and ganglion cells), the retinal nerve fibers and sites of creatine transport into the retina (retinal pigment epithelium, inner retinal blood vessels, and perivascular astrocytes). The CRT was not expressed in Müller cells of any of the species studied. The lack of labeling of Müller cells suggests that neurons are independent of this glial cell in accumulating creatine. During mouse retinal development, expression of the CRT progressively increased throughout the retina until approximately postnatal day 10, with a subsequent decrease. Comparison of the distribution patterns of the CRT in vascular and avascular vertebrate retinas and studies of the mouse retina during development indicate that creatine and phosphocreatine are important for ATP homeostasis.

scholarly journals Extracellular K+ activity changes related to electroretinogram components. II. Rabbit (E-type) retinas.

1985 ◽  
Vol 85 (6) ◽  
pp. 911-931 ◽  
Author(s):  
E Dick ◽  
R F Miller ◽  
S Bloomfield
Keyword(s):  
Retinal Pigment Epithelium ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Müller Cells ◽  
Extracellular Potassium ◽  
Muller Cells ◽  
Positive Component ◽  
Pharmacological Agents ◽  
Slow Piii ◽  
K Activity

Electroretinogram (ERG) and extracellular potassium activity (K+o) measurements were carried out in isolated superfused rabbit eyecup preparations under control conditions and during the application of pharmacological agents that selectively modify the light-responsive retinal network. Light-evoked K+o changes in the rabbit (E-type) retina resemble those previously described in amphibian (I-type) retinas. Different components of the light-evoked K+o changes can be distinguished on the bases of retinal depth, V vs. log I properties, and their responses to pharmacological agents. We find two separable sources of light-evoked increases in extracellular K+: a proximal source and a distal source. The properties of the distal light-evoked K+o increase are consistent with the hypothesis that it initiates a K+-mediated current through Müller cells that is detected as the primary voltage of the electroretinographic b-wave. These experiments also support previous studies indicating that both the corneal-positive component of c-wave and the corneal-negative slow PIII potential result from K+-mediated influences on, respectively, the retinal pigment epithelium and Müller cells.

scholarly journals Cell type-specific complement expression from healthy and diseased retinae

2018 ◽  
Author(s):  
Diana Pauly ◽  
Nicole Schäfer ◽  
Felix Grassmann ◽  
Anna M. Pfaller ◽  
Tobias Straub ◽  
...  
Keyword(s):  
Complement System ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Müller Cells ◽  
Cell Types ◽  
Negative Regulator ◽  
Retinal Cell ◽  
Muller Cells ◽  
Complement Components ◽  
The Individual

AbstractRetinal degeneration is associated with complement system activation, but retinal sources of complement are unknown. Here, we describe the human and murine complement transcriptomes of Müller cells, microglia/macrophages, vascular cells, neurons and retinal pigment epithelium (RPE) in health and disease. All cell populations expressed c1s, c3, cfb, cfp, cfh and cfi. Murine Müller cells contributed the highest amount of complement activators (c1s, c3, cfb). RPE mainly expressed cfh, while cfi and cfp transcripts were most abundant in neurons. The main complement negative regulator in the human retina was cfi, while cfh dominated in the murine retina. Importantly, the expression of c1s, cfb, cfp, cfi increased and that of cfh decreased with aging. Impaired photoreceptor recycling led to an enhanced c3 expression in RPE and to a reduced cfi expression in microglia/macrophages. Expression of complement components was massively upregulated after transient retinal ischemia in murine microglia, Müller cells and RPE. The individual signature of complement expression in distinct murine and human retinal cell types indicates a local, well-orchestrated regulation of the complement system in both species.

scholarly journals Brimonidine Can Prevent In Vitro Hydroquinone Damage on Retinal Pigment Epithelium Cells and Retinal Müller Cells

2016 ◽  
Vol 32 (2) ◽  
pp. 102-108 ◽  
Author(s):  
Claudio Ramírez ◽  
Javier Cáceres-del-Carpio ◽  
Justin Chu ◽  
Joshua Chu ◽  
M. Tarek Moustafa ◽  
...  
Keyword(s):  
Retinal Pigment Epithelium ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Müller Cells ◽  
Muller Cells ◽  
Retinal Pigment Epithelium Cells ◽  
Epithelium Cells
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