Tag Archives: retinitis pigmentosa

Current Perspective on Retinal Remodeling: Implications for Therapeutics

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We have a new paper out of the lab, a perspectives paper on Retinal Remodeling: Implications for Therapeutics. (pdf here).

Authors are Rebecca L. Pfeiffer @BeccaPfeiffer19, and Bryan W. Jones @BWJones.

Abstract: The retinal degenerative diseases retinitis pigmentosa and age-related macular degeneration are a leading cause of irreversible vision loss. Both present with progressive photoreceptor degeneration that is further complicated by processes of retinal remodeling. In this perspective, we discuss the current state of the field of retinal remodeling and its implications for vision-restoring therapeutics currently in development. Here, we discuss the challenges and pitfalls retinal remodeling poses for each therapeutic strategy under the premise that understanding the features of retinal remodeling in totality will provide a basic framework with which therapeutics can interface. Additionally, we discuss the potential for approaching therapeutics using a combined strategy of using diffusible molecules in tandem with other vision-restoring therapeutics. We end by discussing the potential of the retina and retinal remodeling as a model system for more broadly understanding the progression of neurodegeneration across the central nervous system.

Müller Cell Metabolic Chaos During Retinal Degeneration

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We have a new publication out (direct link, open access), Müller Cell Metabolic Chaos During Retinal Degeneration authored by Rebecca Pfeiffer, Robert Marc, Mineo Kondo, Hiroko Terasaki and Bryan W. Jones.

Abstract:

Müller cells play a critical role in retinal metabolism and are among the first cells to demonstrate metabolic changes in retinal stress or disease. The timing, extent, regulation, and impacts of these changes are not yet known. We evaluated metabolic phenotypes of Müller cells in the degenerating retina.

Retinas harvested from wild-type (WT) and rhodopsin Tg P347L rabbits were fixed in mixed aldehydes and resin embedded for computational molecular phenotyping (CMP). CMP facilitates small molecule fingerprinting of every cell in the retina, allowing evaluation of metabolite levels in single cells.

CMP revealed signature variations in metabolite levels across Müller cells from TgP347L retina. In brief, neighboring Müller cells demonstrated variability in taurine, glutamate, glutamine, glutathione, glutamine synthetase (GS), and CRALBP. This variability showed no correlation across metabolites, implying the changes are functionally chaotic rather than simply heterogeneous. The inability of any clustering algorithm to classify Müller cell as a single class in the TgP347L retina is a formal proof of metabolic variability in the present in degenerating retina.

Although retinal degeneration is certainly the trigger, Müller cell metabolic alterations are not a coherent response to the microenvironment. And while GS is believed to be the primary enzyme responsible for the conversion of glutamate to glutamine in the retina, alternative pathways appear to be unmasked in degenerating retina. Somehow, long term remodeling involves loss of Müller cell coordination and identity, which has negative implications for therapeutic interventions that target neurons alone.

Progressive Retinal Remodeling In A Transgenic Rabbit Model Of Retinitis Pigmentosa

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This poster was presented today, May 2th at the 2016 Association for Research in Vision and Opthalmology (ARVO) meetings in Seattle, Washington by Rebecca L. Pfeiffer, Bryan W. Jones, and Robert E. Marc.

Posterboard #: D0246

Abstract Number: 2256 – D0246

Author Block: Rebecca L. Pfeiffer1,2 , Bryan W. Jones1,2 , Robert E. Marc1,2 
1 Ophthalmology, University of Utah, Salt Lake City, Utah, United States; 2 Interdepartmental Program in Neuroscience, University of Utah, Salt Lake City, Utah, United States

Purpose:Retinal degenerations are a collection of neural disorders, usually precipitated by photoreceptor degeneration. All display progressive metabolic alterations and neural loss following the death of the photoreceptors. Although it has been demonstrated that the metabolism of Müller cells (MCs) is drastically altered in degeneration, the full impact of these changes on surrounding neurons and long-term characterization of remodeling was previously unavailable, due to short lifespans of model organisms.

Methods:Retinal samples were collected from WT and Tg P347L rabbits at ages ranging from 3 months to 6 years. Following enucleation, retinas were divided into fragments and incubated for 10 minutes at 35 degrees C in D-isomers of Glutamate (dE), Glutamine (dQ), or Aspartate (dD) and Ames-bicarbonate medium to explore retinal transport capabilities at each stage of degeneration. Retinas were then fixed in mixed aldehyde buffer and processed for transmission electron microscope connectomics, immunocytochemistry for a range of macromolecules, and computational molecular phenotyping for small molecules (CMP) (J Comp Neurol. 464:1, 2003).

Results:CMP reveals that single metabolic phenotype of MCs splits and diverges into many phenotypes continuously throughout degeneration. Further, all neuronal classes continue to die throughout degeneration. By 6 years, over 90% of neurons are lost, and the remaining glutamatergic neurons have altered metabolic signatures with a large increase in aspartate levels, at times exceeding glutamate. Transport of the D-isomers indicates that glutamate transport capabilities remain intact until the latest stages of degeneration. This may not be true of their GABA transporters.

Conclusions:These results suggest three main conclusions. First, retinal remodeling in degeneration is relentlessly progressive long after all photoreceptors have disappeared. Second, cell types previously thought to remain after degeneration onset, such as ganglion cells, will also ultimately die and the cells not lost often will change their metabolism. The consequence of this metabolic change in neurons is not yet fully explored. Third, the persistent robust glutamate transport capabilities of Müller cells indicate Müller cells can metabolize glutamate despite the massive loss of glutamine synthetase activity, likely unmasking alternate metabolic pathways.

Retinal Remodeling in Human Retinitis Pigmentosa

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We have a new publication out (Direct Link, Free Open Access), Retinal Remodeling in Human Retinitis Pigmentosa authored by Bryan W. Jones, Rebecca Pfeiffer, Drew Ferrell, Carl Watt, Michael Marmor and Robert Marc.

Abstract: Retinitis Pigmentosa (RP) in the human is a progressive, currently irreversible neural degenerative disease usually caused by gene defects that disrupt the function or architecture of the photoreceptors. While RP can initially be a disease of photoreceptors, there is increasing evidence that the inner retina becomes progressively disorganized as the outer retina degenerates. These alterations have been extensively described in animal models, but remodeling in humans has not been as well characterized. This study, using computational molecular phenotyping (CMP) seeks to advance our understanding of the retinal remodeling process in humans. We describe cone mediated preservation of overall topology, retinal reprogramming in the earliest stages of the disease in retinal bipolar cells, and alterations in both small molecule and protein signatures of neurons and glia. Furthermore, while Müller glia appear to be some of the last cells left in the degenerate retina, they are also one of the first cell classes in the neural retina to respond to stress which may reveal mechanisms related to remodeling and cell death in other retinal cell classes. Also fundamentally important is the finding that retinal network topologies are altered. Our results suggest interventions that presume substantial preservation of the neural retina will likely fail in late stages of the disease. Even early intervention offers no guarantee that the interventions will be immune to progressive remodeling. Fundamental work in the biology and mechanisms of disease progression are needed to support vision rescue strategies.

Webvision Chapter: Retinal Degeneration, Remodeling and Plasticity

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We have published a new chapter in Webvision, Retinal Degeneration, Remodeling and Plasticity that covers the history of the study of retinal degenerations and some of the implications for vision rescue.  Authors are Bryan W. Jones, Rebecca L. Pfeiffer and Robert E. Marc.  It will, like other Webvision chapters evolve over time, which is the whole point of Webvision, but we hope it will generate some discussion.

Metabolic Changes Associated With Müller Cells In A Transgenic Rabbit Model Of Retinal Degeneration

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Retina-RLP

This abstract was presented today at the 2014 Association for Research in Vision and Opthalmology (ARVO) meetings in Orlando, Florida by  Rebecca L. Pfeiffer, Bryan W. Jones and Robert E. Marc.

Purpose: Müller cells play a central role in retinal metabolism via the glutamate cycle. During retinal degeneration Müller cells are among the first to demonstrate changes, reflected in alterations of metabolic signatures and morphology. The timing, extent and regulation of these changes is not fully characterized. To address this issue, we evaluated Müller cell metabolic phenotypes at multiple stages of retinal remodeling.

Methods: Samples were collected post-mortem from both WT and P347L rabbits. The retinas were then divided into fragments, fixed in buffered aldehydes, and embedded in epoxy resins. Tissues were sectioned at 200nm followed by classification with computational molecular phenotyping (CMP) using an array of small and macromolecular signatures (aspartate (D), glutamate (E), glycine (G), glutamine (Q), glutathione (J), GABA (yy), taurine (T), CRALBP, Glutamine Synthetase (GS), and GFAP). Levels of amino acid or protein were quantified by selecting a region of interest either within the Müller cell population or surrounding neurons and evaluating the intensity of the signal within that region.

Results: CMP reveals overall decreases in GS levels over the course of degeneration. Of notable importance, we saw that in regions of near complete photoreceptor loss neighboring Müller cells may express independent variation in metabolic signatures of E, Q, and GS. Also observed in these Müller cells, ratios of GS:E and GS:Q are not consistent with the ratios seen in WT retina. These results are inconsistent with the current models of both E to Q metabolism and microenvironment regulation of Müller cell phenotypes.

Conclusions: These observations indicate two conclusions. First, although the degenerate state of the retina is the likely trigger inducing Müller cells to express altered metabolic signatures, the rate at which the metabolic state changes is not purely a product of the surrounding environment, but also a stochastic change within individual Müller cells. Second, although it is commonly accepted that GS is the primary enzyme which converts Q to E as part of the glutamate cycle, in degenerate retina alternative pathways may be utilized following decrease in GS.

Support: NIH EY02576 (RM), NIH EY015128 (RM), NSF 0941717 (RM), NIH EY014800 Vision Core (RM), RPB CDA (BWJ), Thome AMD Grant (BWJ).