Evidence for glutamate-mediated excitotoxic mechanisms during photoreceptor degeneration in the rd1 mouse retina

PURPOSE: Kinetic studies of photoreceptor cell death in the retinal degeneration (rd1) mouse model suggest that photoreceptor degeneration could result from cumulative damage. Since alterations in glutamate metabolism have been described in different models of retinitis pigmentosa, we investigated in the present work whether changes in glutamate turnover occur in the degenerating rd1 retina and whether glutamate-mediated excitotoxic mechanisms may contribute to rod photoreceptor death in this model. METHODS: Free amino acid levels were quantified in rd1 and wild-type retinas using an amino acid analyzer selecting times corresponding to early, intermediate, and terminal phases of rod photoreceptor degeneration. Reverse transcription-polymerase chain reaction (RT-PCR) was used to compare the mRNA expression levels of the glial L-glutamate/L-aspartate transporter GLAST, glutamine synthetase (GS), and vimentin, a marker for retinal glia, between rd1 and wild-type mouse retinas. 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), an antagonist of both AMPA and kainate subtypes of ionotropic glutamate receptors, was then daily administered from postnatal day 3 (PN3) to PN21 to rd1 mice while control rd1 mice received only physiological saline solution (7 per treatment). At PN22, the respective numbers of surviving rods i

A Generalized Typicality for Abstract Alphabets

A new notion of typicality for arbitrary probability measures on standard Borel spaces is proposed, which encompasses the classical notions of weak and strong typicality as special cases. Useful lemmas about strong typical sets, including conditional typicality lemma, joint typicality lemma, and packing and covering lemmas, which are fundamental tools for deriving many inner bounds of various multi-terminal coding problems, are obtained in terms of the proposed notion. This enables us to directly generalize lots of results on finite alphabet problems to general problems involving abstract alphabets, without any complicated additional arguments. For instance, quantization procedure is no longer necessary to achieve such generalizations. Another fundamental lemma, Markov lemma, is also obtained but its scope of application is quite limited compared to others. Yet, an alternative theory of typical sets for Gaussian measures, free from this limitation, is also developed. Some remarks on a possibility to generalize the proposed notion for sources with memory are also given.Comment: 44 pages; submitted to IEEE Transactions on Information Theor

The matricial relaxation of a linear matrix inequality

Given linear matrix inequalities (LMIs) L_1 and L_2, it is natural to ask: (Q1) when does one dominate the other, that is, does L_1(X) PsD imply L_2(X) PsD? (Q2) when do they have the same solution set? Such questions can be NP-hard. This paper describes a natural relaxation of an LMI, based on substituting matrices for the variables x_j. With this relaxation, the domination questions (Q1) and (Q2) have elegant answers, indeed reduce to constructible semidefinite programs. Assume there is an X such that L_1(X) and L_2(X) are both PD, and suppose the positivity domain of L_1 is bounded. For our "matrix variable" relaxation a positive answer to (Q1) is equivalent to the existence of matrices V_j such that L_2(x)=V_1^* L_1(x) V_1 + ... + V_k^* L_1(x) V_k. As for (Q2) we show that, up to redundancy, L_1 and L_2 are unitarily equivalent. Such algebraic certificates are typically called Positivstellensaetze and the above are examples of such for linear polynomials. The paper goes on to derive a cleaner and more powerful Putinar-type Positivstellensatz for polynomials positive on a bounded set of the form {X | L(X) PsD}. An observation at the core of the paper is that the relaxed LMI domination problem is equivalent to a classical problem. Namely, the problem of determining if a linear map from a subspace of matrices to a matrix algebra is "completely positive".Comment: v1: 34 pages, v2: 41 pages; supplementary material is available in the source file, or see http://srag.fmf.uni-lj.si

Translational Retinal Research and Therapies.

The following review summarizes the state of the art in representative aspects of gene therapy/translational medicine and evolves from a symposium held at the School of Veterinary Medicine, University of Pennsylvania on November 16, 2017 honoring Dr. Gustavo Aguirre, recipient of ARVO's 2017 Proctor Medal. Focusing on the retina, speakers highlighted current work on moving therapies for inherited retinal degenerative diseases from the laboratory bench to the clinic

2020-2021 Georgia Southern University Women\u27s Golf Statistics

Georgia Southern Eagles 2020-21 Women\u27s Golf Statistic

Present Molecular Limitations of ON-Bipolar Cell Targeted Gene Therapy

Recent studies have demonstrated the safety and efficacy of ocular gene therapy based on adeno-associated viral vectors (AAVs). Accordingly, a surge in promising new gene therapies is entering clinical trials, including the first optogenetic therapy for vision restoration. To date, optogenetic therapies for vision restoration target either the retinal ganglion cells (GCs) or presynaptic ON-bipolar cells (OBCs). Initiating light responses at the level of the OBCs has significant advantages over optogenetic activation of GCs. For example, important neural circuitries in the inner retina, which shape the receptive fields of GCs, remain intact when activating the OBCs. Current drawbacks of AAV-mediated gene therapies targeting OBCs include (1) a low transduction efficiency, (2) off-target expression in unwanted cell populations, and (3) a poor performance in human tissue compared to the murine retina. Here, we examined side-by-side the performance of three state-of-the art AAV capsid variants, AAV7m8, AAVBP2, and AAV7m8(Y444F) in combination with the 4xGRM6-SV40 promoter construct in the healthy and degenerated mouse retina and in human post-mortem retinal explants. We find that (1) the 4xGRM6-SV40 promoter is not OBC specific, (2) that all AAV variants possess broad cellular transduction patterns, with differences between the transduction patterns of capsid variants AAVBP2 and AAV7m8 and, most importantly, (3) that all vectors target OBCs in healthy tissue but not in the degenerated rd1 mouse model, potentially limiting the possibilities for an OBC-targeted optogenetic therapy for vision restoration in the blind

Functional Availability of ON-Bipolar Cells in the Degenerated Retina: Timing and Longevity of an Optogenetic Gene Therapy

Degenerative diseases of the retina are responsible for the death of photoreceptors and subsequent loss of vision in patients. Nevertheless, the inner retinal layers remain intact over an extended period of time, enabling the restoration of light sensitivity in blind retinas via the expression of optogenetic tools in the remaining retinal cells. The chimeric Opto-mGluR6 protein represents such a tool. With exclusive ON-bipolar cell expression, it combines the light-sensitive domains of melanopsin and the intracellular domains of the metabotropic glutamate receptor 6 (mGluR6), which naturally mediates light responses in these cells. Albeit vision restoration in blind mice by Opto-mGluR6 delivery was previously shown, much is left to be explored in regard to the effects of the timing of the treatment in the degenerated retina. We performed a functional evaluation of Opto-mGluR6-treated murine blind retinas using multi-electrode arrays (MEAs) and observed long-term functional preservation in the treated retinas, as well as successful therapeutical intervention in later stages of degeneration. Moreover, the treatment decreased the inherent retinal hyperactivity of the degenerated retinas to levels undistinguishable from healthy controls. Finally, we observed for the first time micro electroretinograms (mERGs) in optogenetically treated animals, corroborating the origin of Opto-mGluR6 signalling at the level of mGluR6 of ON-bipolar cells