Transmission de l'information visuelle : codage et restauration visuelle
Chef d'équipe : Olivier Marre
INTRODUCTION | EQUIPE | PRESENTATION | DOMAINE DE RECHERCHE | PUBLICATIONS | ILS NOUS SOUTIENNENT
Introduction
Notre équipe étudie la transmission de l'information visuelle en condition normale ou pathologique. Nous développons et testons également de nouvelles stratégies thérapeutiques pour prévenir la perte de vision et restaurer la vision chez les patients aveugles.
Présentation
Notre deuxième objectif est de développer des solutions pour restaurer la vision chez des patients aveugles, soit en utilisant des prothèses rétiniennes, soit avec une thérapie optogénétique. Pour les prothèses rétiniennes, nous contribuons au développement de nouveaux matériaux pour les électrodes (diamant, graphène). Les deux approches sont validées sur des animaux aveugles (souris, rat) puis testées sur le primate non-humain. Nous pouvons ainsi mesurer si la sortie de la rétine est modifiée par ces stratégies, afin de comprendre ce qui sera envoyé au cerveau. Nous pouvons aussi enregistrer l'activité corticale in vivo pour démontrer l'efficacité fonctionnelle de ces traitements. Le développement de la thérapie optogénétique est un projet tranversal avec les équipes de Deniz Dalkara et de Jens Duebel.
Enfin, notre troisème ambition est de développer des stratégies neuroprotectives. Nous caractérisons les phénotypes rétiniens dans des modèles animaux de différentes maladies de la rétine (OTX2, COL4A1, Usher, etc.) pour ensuite tester différentes stratégies thérapeutiques. Nous sommes particulièrement intéressés par l'effet délétère de la lumière sur la rétine sa prévention potentielle par des filtres de couleur ou des antioxidants comme la taurine. Dans des modèles cellulaires de la dégénérescence maculaire liée à l'age, nous déterminons quelles couleurs sont les plus toxiques. Nous avons aussi montré que la dépletion en taurine peut mener à un dégénérescence des cones et des cellules ganglionnaires dans différents modèles animaux. Nous cherchons actuellement à comprendre les mécanismes moléculaires et cellulaires induisant cette dégénérescence liée à des facteurs environnementaux.
Domaines de recherche
- Neuroscience, vision, ophtalmologie, neuroscience computationnelle, pharmacologie, thérapie optogénétique, thérapie génique, dispositif médical, prothèse rétinienne, prévention des maladies, régime, nutricament, taurine, phototoxicité, handicap, transmission de l'information rétinienne, modèles animaux, glaucome, syndrome de Usher, dégénérescence maculaire liée à l'âge, rétinopathie diabétique.
Team Leader
Publications
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Liste actualisée et complète par scientifique
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2020
Phototoxic damage to cone photoreceptors can be independent of the visual pigment: the porphyrin hypothesis
Mélanie Marie, Valérie Forster, Stéphane Fouquet, Pascal Berto, Coralie Barrau, Camille Ehrismann, José-Alain Sahel, Gilles Tessier & Serge Picaud
Cell Death & Disease volume 11, Article number: 711
VEGF is an autocrine/paracrine neuroprotective factor for injured retinal ganglion neurons.
Froger N, Matonti F, Roubeix C, Forster V, Ivkovic I, Brunel N, Baudouin C, Sahel JA, Picaud S. Sci Rep. 2020 Jul 24;10(1):12409. doi: 10.1038/s41598-020-68488-z. PMID: 32710087 Free PMC article.Towards optogenetic vision restoration with high resolution.
Ferrari U, Deny S, Sengupta A, Caplette R, Trapani F, Sahel JA, Dalkara D, Picaud S, Duebel J, Marre O. PLoS Comput Biol. 2020 Jul 15;16(7):e1007857. doi: 10.1371/journal.pcbi.1007857. eCollection 2020 Jul. PMID: 32667921 Free PMC article.Functional ultrasound imaging of deep visual cortex in awake nonhuman primates.
Blaize K, Arcizet F, Gesnik M, Ahnine H, Ferrari U, Deffieux T, Pouget P, Chavane F, Fink M, Sahel JA, Tanter M, Picaud S. Proc Natl Acad Sci U S A. 2020 Jun 23;117(25):14453-14463. doi: 10.1073/pnas.1916787117. Epub 2020 Jun 8. PMID: 32513717 Free PMC article.Impact of the COVID-19 lockdown on basic science research in ophthalmology: the experience of a highly specialized research facility in France.
Nassisi M, Audo I, Zeitz C, Varin J, Wohlschlegel J, Smirnov V, Santiard-Baron D, Picaud S, Sahel JA. Eye (Lond). 2020 Jul;34(7):1187-1188. doi: 10.1038/s41433-020-0944-7. Epub 2020 May 7. PMID: 32382143 Free PMC article. No abstract available.Impedance spectroscopy study of the retinal pigment epithelium: Application to the monitoring of blue light exposure effect on A2E-loaded in-vitro cell cultures.
Boutzen J, Valet M, Alviset A, Fradot V, Rousseau L, Français O, Picaud S, Lissorgues G. Biosens Bioelectron. 2020 Aug 1;161:112180. doi: 10.1016/j.bios.2020.112180. Epub 2020 Apr 5. PMID: 32365009PRIMA subretinal wireless photovoltaic microchip implantation in non-human primate and feline models.
Muqit MMK, Hubschman JP, Picaud S, McCreery DB, van Meurs JC, Hornig R, Buc G, Deterre M, Nouvel-Jaillard C, Bouillet E, Fovet CM, Hantraye P, Sahel J, Martel JN, Le Mer Y. PLoS One. 2020 Apr 8;15(4):e0230713. doi: 10.1371/journal.pone.0230713. eCollection 2020. PMID: 32267845 Free PMC article.IL-1β induces rod degeneration through the disruption of retinal glutamate homeostasis.
Charles-Messance H, Blot G, Couturier A, Vignaud L, Touhami S, Beguier F, Siqueiros L, Forster V, Barmo N, Augustin S, Picaud S, Sahel JA, Rendon A, Grosche A, Tadayoni R, Sennlaub F, Guillonneau X. J Neuroinflammation. 2020 Jan 3;17(1):1. doi: 10.1186/s12974-019-1655-5. PMID: 31900165 -
2019
Functional efficacy of stimulations with a photovoltaic sub-retinal prosthesis in non-human primates.
Prévot P.H., Gehere K., Arcizet F., Akolkar H., Khoei M. A., Blaize K., Oubari O., Daye P., Lanoë M., Valet M., Dalouz S., Langlois P., Esposito E., Forster V., Dubus E., Wattiez N., Brazhnikova E., Nouvel-Jaillard C., LeMer Y., Demilly J., Fovet C.M., Hantraye P., Weissenburger M., Lorach H., Bouillet E., Deterre M., Hornig R., Buc G., Sahel J.A., Chenegros G., Pouget P*., Benosman R*., Picaud S*. (* Co-last authors)
Nature Biomed. Eng. (2019, sous presse)Functional ultrasound imaging of the brain reveals propagation of task-related brain activity in behaving primates.
Dizeux A, Gesnik M, Ahnine H, Blaize K, Arcizet F, Picaud S, Sahel JA, Deffieux T, Pouget P, Tanter M
Nat Commun. 2019 Mar 28;10(1):1400.A biophysical model explains the spontaneous bursting behavior in the developing retina.
Matzakos-Karvouniari D, Gil L, Orendorff E, Marre O, Picaud S, Cessac B.
Sci Rep. (2019) 9:1859. doi: 10.1038/s41598-018-38299-4. -
2018
High‐Frequency Stimulation of Normal and Blind Mouse Retinas Using TiO2 Nanotubes
Ronzani C, Cottineau T, Gonzalez‐Valls I, Keller V, Picaud S, Keller N, Roux MJ
Advanced Functional Materials (2018) 28, 1804639Taurine Depletion Causes ipRGC Loss and Increases Light-Induced Photoreceptor Degeneration.Taurine Depletion Causes ipRGC Loss and Increases Light-Induced Photoreceptor Degeneration.
García-Ayuso D, Di Pierdomenico J, Hadj-Said W, Marie M, Agudo-Barriuso M, Vidal-Sanz M, Picaud S, Villegas-Pérez MP.
Invest Ophthalmol Vis Sci. (2018) 59:1396-1409.Light action spectrum on oxidative stress and mitochondrial damage in A2E-loaded retinal pigment epithelium cells.
Marie M, Bigot K, Angebault C, Barrau C, Gondouin P, Pagan D, Fouquet S, Villette T, Sahel JA, Lenaers G, Picaud S.
Cell Death Dis. (2018) 9:287. doi: 10.1038/s41419-018-0331-5Noninvasive gene delivery to foveal cones for vision restoration.
Khabou H, Garita-Hernandez M, Chaffiol A, Reichman S, Jaillard C, Brazhnikova E, Bertin S, Forster V, Desrosiers M, Winckler C, Goureau O, Picaud S, Duebel J, Sahel JA, Dalkara D.
JCI Insight. (2018) 3. pii: 96029.Cone degeneration is triggered by the absence of USH1 proteins but prevented by antioxidant treatments.
Trouillet A, Dubus E, Dégardin J, Estivalet A, Ivkovic I, Godefroy D, García-Ayuso D, Simonutti M, Sahly I, Sahel JA, El-Amraoui A, Petit C, Picaud S.
Sci Rep. (2018) 8:1968. doi: 10.1038/s41598-018-20171-0Nonlinear decoding of a complex movie from the mammalian retina.
Botella-Soler V, Deny S, Martius G, Marre O, Tkačik G.
PLoS Comput Biol. 2018 May 10;14(5):e1006057.A spike sorting toolbox for up to thousands of electrodes validated with ground truth recordings in vitro and in vivo.
Yger P, Spampinato GL, Esposito E, Lefebvre B, Deny S, Gardella C, Stimberg M, Jetter F, Zeck G, Picaud S, Duebel J, Marre O.
Elife. 2018 Mar 20;7.Blindfold learning of an accurate neural metric.
Gardella C, Marre O, Mora T.
Proc Natl Acad Sci U S A. 2018 Mar 27;115(13):3267-3272.Closed-Loop Estimation of Retinal Network Sensitivity by Local Empirical Linearization.
Ferrari U, Gardella C, Marre O, Mora T.
eNeuro. 2018 Jan 23;4(6).Toward a unified theory of efficient, predictive, and sparse coding.
Chalk M, Marre O, Tkačik G.
Proc Natl Acad Sci U S A. 2018 Jan 2;115(1):186-191. -
2017
Multiplexed computations in retinal ganglion cells of a single type
Deny S, Ferrari U, Mace E, Yger P, Caplette R, Picaud S, Tkacik G, Marre O
Nature Communications, 8:1964A new promoter allows optogenetic vision restoration with enhanced sensitivity in macaque retina.
Chaffiol, A., Caplette, R. Jaillard, C., Brazhnikova, E., Desrosiers, M., Dubus, E., Duhamel, L., Macé, E., Marre, O., Benoit, P., Hantraye, P., Bemelmans, A. P., Bamberg, E., Duebel, J., Sahel, J. A., Picaud, S., Dalkara, D.
Molecular Therapy (2017) 25: 2546-25603D functional ultrasound imaging of the cerebral visual system in rodents.
Gesnik M, Blaize K, Deffieux T, Gennisson JL, Sahel JA, Fink M, Picaud S, Tanter M
Neuroimage. 2017 Apr 1;149:267-274Col4a1 mutation generates vascular abnormalities correlated with neuronal damage in a mouse model of HANAC syndrome.
Trouillet A, Lorach H, Dubus E, El Mathari B, Ivkovic I, Dégardin J, Simonutti M, Paques M, Guillonneau X, Sennlaub F, Sahel JA, Ronco P, Plaisier E, Picaud S.
Neurobiol Dis. 2017 Jan 3;100:52-61.Random versus maximum entropy models of neural population activity.
Ferrari U, Obuchi T, Mora T
Phys Rev E. Apr;95(4-1):042321. -
2016
Tractable Method for Describing Complex Couplings between Neurons and Population Rate
Gardella C, Marre O, Mora T.
eNeuro. 2016 Aug 18;3(4).Red-shifted channelrhodopsin stimulation restores light responses in blind mice, macaque retina, and human retina
Sengupta, A., Chaffiol, A., Mace, E., Caplette, R., Desrosiers, M., Lampic, M., Forster, V., Marre, O., Lin, J.Y., Sahel, J.A., Picaud, S., Dalkara, D. & Duebel, J.
EMBO Mol Med (2016).Fast and accurate spike sorting in vitro and in vivo for up to thousands of electrodes
P. Yger, G. L. B. Spampinato, E. Esposito, B. Lefebvre, S. Deny, C. Gardella, M. Stimberg, F. Jetter, G. Zeck, S. Picaud, J. Duebel and O. Marre.
(2016), BioRxiv, 067843Nonlinear decoding of a complex movie from the mammalian retina.
Botella-Soler V, Deny S, Marre O, Tkacik G. (2016) arXiv: 1605.03373 [q-bio.NC]Relevant sparse codes with variational information bottleneck Neural Information Processing Systems
Chalk M, Marre O, Tkacik G. (2016) in press. arXiv: 1605.07332 [q-bio.NC]A tractable method for describing complex couplings between neurons and population rate
Gardella C, Marre O, Mora T. (2016) eNeuro 3(4) e0160-15Study of retinal alterations in a high fat diet-induced type ii diabetes rodent: Meriones shawi
Hammoum I, Mbarek S, Dellaa A, Dubus E, Baccouche B, Azaiz R, Charfeddine R, Picaud S, Ben Chaouacha-Chekir R.
Acta Histochem. 2016 Jun 2. pii: S0065-1281(16)30090-3. -
2015
Genotypic and phenotypic characterization of P23H line 1 rat model.
Orhan E, Dalkara D, Neuillé M, Lechauve C, Michiels C, Picaud S, Léveillard T, Sahel JA, Naash MI, Lavail MM, Zeitz C, Audo I.
PLoS One. 2015 May 26;10(5):e0127319. doi: 10.1371/journal.pone.0127319. eCollection 2015.
PMID:260098933D-nanostructured boron-doped diamond for microelectrode array neural interfacing.
Piret G, Hébert C, Mazellier JP, Rousseau L, Scorsone E, Cottance M, Lissorgues G, Heuschkel MO, Picaud S, Bergonzo P, Yvert B.
Biomaterials. 2015 Jun;53:173-83. doi: 10.1016/j.biomaterials.2015.02.021. Epub 2015 Mar 13.
PMID:25890717Sildenafil Acutely Decreases Visual Responses in ON and OFF Retinal Ganglion Cells.
Martins J, Kolomiets B, Caplette R, Sahel JA, Castelo-Branco M, Ambrósio AF, Picaud S.
Invest Ophthalmol Vis Sci. 2015 Apr 1;56(4):2639-48. doi: 10.1167/iovs.14-15964.PMID:2581400Thermodynamics and signatures of criticality in a network of neurons.
Tkacik G, Mora T, Marre O, Amodei D, Palmer SE, Berry MJ 2nd, Bialek W (2015). PNAS, Sep 1.Precise decoding of dynamical motion from a large retinal population.
Marre, O., Tkacik, G., Simmons, K., Mora, T., Berry, M.J. (2015). PloS Computational Biology, Jul 1;11(7):e1004304.Predictive information in a sensory population.
Palmer, S.E., Marre, O., Berry II, M.J., Bialek, W. (2015). PNAS, Jun 2;112(22):6908-13.What can asynchronous event-based acquisition add to pattern recognition?
Akolkar, H., Meyer, C., Clady, X., Marre, O., Bartolozzi, C., Panzeri,S., Benosman, R. (2015). Neural Computation, 27 561-593Dynamical criticality in the collective activity of a population of retinal neurons.
Mora, T., Deny, S., Marre O. (2015). Physical Review Letters, 114, 078105 -
2014
Determination of morphological, biometric and biochemical susceptibilities in healthy eurasier dogs with suspected inherited glaucoma.
Boillot T, Rosolen SG, Dulaurent T, Goulle F, Thomas P, Isard PF, Azoulay T, Lafarge-Beurlet S, Woods M, Lavillegrand S, Ivkovic I, Neveux N, Sahel JA, Picaud S, Froger N.
PLoS One. 2014 Nov 7;9(11):e111873. doi: 10.1371/journal.pone.0111873. eCollection 2014.Targeting Channelrhodopsin-2 to ON-bipolar Cells With Vitreally Administered AAV Restores ON and OFF Visual Responses in Blind Mice.
Macé E, Caplette R, Marre O, Sengupta A, Chaffiol A, Barbe P, Desrosiers M, Bamberg E, Sahel JA, Picaud S, Duebel J, Dalkara D. Mol Ther. 2014 Aug 6. doi: 10.1038/mt.2014.154. [Epub ahead of print]Taurine: the comeback of a neutraceutical in the prevention of retinal degenerations
Froger N, Moutsimilli L, Cadetti L, Jammoul F, Wang QP, Fan Y, Gaucher D, Rosolen SG, Neveux N, Cynober L, Sahel JA, Picaud S.
Prog Retin Eye Res. 2014 Jul;41:44-63. doi: 10.1016/j.preteyeres.2014.03.001. Epub 2014 Apr 8.Retinal prostheses: clinical results and future challenges.
Picaud S, Sahel JA.
C R Biol. 2014 Mar;337(3):214-22. doi: 10.1016/j.crvi.2014.01.001. Epub 2014 Feb 26. Review.Cell specific electrodes for neuronal network reconstruction and monitoring.
Bendali A, Bouguelia S, Roupioz Y, Forster V, Mailley P, Benosman R, Livache T, Sahel JA, Picaud S.
Analyst. 2014 Jul 7;139(13):3281-9. doi: 10.1039/c4an00048j.Distinctive glial and neuronal interfacing on nanocrystalline diamond.
Bendali A, Agnès C, Meffert S, Forster V, Bongrain A, Arnault JC, Sahel JA, Offenhäusser A, Bergonzo P, Picaud S.
PLoS One. 2014 Mar 24;9(3):e92562. doi: 10.1371/journal.pone.0092562. eCollection 2014.Graded Otx2 activities demonstrate dose-sensitive eye and retina phenotypes.
Bernard C, Kim HT, Torero Ibad R, Lee EJ, Simonutti M, Picaud S, Acampora D, Simeone A, Di Nardo AA, Prochiantz A, Moya KL, Kim JW.
Hum Mol Genet. 2014 Apr 1;23(7):1742-53. doi: 10.1093/hmg/ddt562. Epub 2013 Nov 13.Boron doped diamond biotechnology: from sensors to neurointerfaces.
Hébert C, Scorsone E, Bendali A, Kiran R, Cottance M, Girard HA, Degardin J, Dubus E, Lissorgues G, Rousseau L, Mailley P, Picaud S, Bergonzo P.
Faraday Discuss. 2014;172:47-59. doi: 10.1039/c4fd00040d. Epub 2014 Sep 26.Searching for collective behaviour in a network of real neurons.
Tkacik, G., Marre, O., Amodei, D., Schneidman, E., Bialek, W, Berry M.J., (2014). PloS Computational Biology Jan;10(1):e1003408 -
2013
Sahel JA, Leveillard T, Picaud S, Dalkara D, Marazova K, Safran A, Paques M, Duebel J, Roska B, Mohand-Said S. Functional rescue of cone photoreceptors in retinitis pigmentosa.
Graefes Arch Clin Exp Ophthalmol. 2013 Jul;251(7):1669-77.Bernard C, Kim HT, Torero Ibad R, Lee EJ, Simonutti M, Picaud S, Acampora D,Simeone A, Di Nardo AA, Prochiantz A, Moya KL, Kim JW. Graded Otx2 activities demonstrate dose-sensitive eye and retina phenotypes. Hum Mol Genet. 2013 Nov 28.
Linderholm P, Guyomard JL, Djilas M, Salzmann J, Simonutti M, Sahel JA, Safran AB, Renaud P, Picaud S. Long-term in vivo impedance changes of subretinal microelectrodes implanted in dystrophic P23H rats. Int J Artif Organs. 2013 Oct 3;36(9):612-9.Lorach H, Marre O, Sahel JA, Benosman R, Picaud S. Neural stimulation for visual rehabilitation: advances and challenges.
J Physiol Paris. 2013 Nov;107(5):421-31.Arnault E, Barrau C, Nanteau C, Gondouin P, Bigot K, Vienot F, Gutman E, Fontaine V, Villette T, Cohen-Tannoudji D, Sahel JA, Picaud S. Phototoxic action spectrum on a retinal pigment epithelium model of age-related macular degeneration exposed to sunlight normalized conditions. PLoS One. 2013 Aug 23;8(8):e71398.
Bendali A, Hess LH, Seifert M, Forster V, Stephan AF, Garrido JA, Picaud S.
Purified neurons can survive on peptide-free graphene layers. Adv Healthc Mater. 2013 Jul;2(7):929-33.Bendali A, Lorach H, Djilas M, Marre O, Bensoman R, Rousseau L, Lissorgues G, Scorsone E, Bergonzo P, Garrido JA, Sahel JA, Picaud S. Restoring vision in blind patients following photoreceptor degeneration: clinical results and future challenges. Biol Aujourdhui. 2013;207(2):123-32.
Roska B, Busskamp V, Sahel JA, Picaud S. Retinitis pigmentosa: eye sight restoration by optogenetic therapy.
Biol Aujourdhui. 2013; 207(2):109-21.Froger N, Jammoul F, Gaucher D, Cadetti L, Lorach H, Degardin J, Pain D, Dubus E, Forster V, Ivkovic I, Simonutti M, Sahel JA, Picaud S. Taurine is a crucial factor to preserve retinal ganglion cell survival. Adv Exp Med Biol. 2013;775:69-83.
Girmens JF, Picaud S, Sahel JA. Therapeutic innovation in AMD and other retinal diseases.
Rev Prat. 2013 Jan;63(1):68-73. ReviewThe simplest maximum entropy model for collective behavior in a neural network.
Tkacik, G., Marre, O., Mora, T., Amodei, D., Berry M.J., Bialek, W (2013). Journal of Statistical mechanics, P03011.Spatio-temporal spike trains analysis for large-scale networks using the maximum entropy principle and a Monte-Carlo method.
Nasser, H., Marre, O., Cessac, B. (2013). Journal of Statistical Mechanics, P03006. -
2011
Bougrid A, Claudepierre T, Picaud S, Ayad G, Mornet D, Dorbani-Mamine L, Rendon A, Darbeida H.Expression of Dystrophins and the Dystrophin-Associated-Protein Complex by Pituicytes in Culture.Neurochem Res. 2011 Apr 12. [Epub ahead of print]Ibad RT, Rheey J, Mrejen S, Forster V, Picaud S, Prochiantz A, Moya KL.Otx2 Promotes the Survival of Damaged Adult Retinal Ganglion Cells and Protects against Excitotoxic Loss of Visual Acuity In Vivo.J Neurosci. 2011 Apr 6;31(14):5495-5503.
Kasi H, Bertsch A, Guyomard JL, Kolomiets B, Picaud S, Pelizzone M, Renaud P.Simulations to study spatial extent of stimulation and effect of electrode-tissue gap in subretinal implants.Med Eng Phys. 2011 Feb 25. [Epub ahead of print]
Fournier I, Gaucher D, Chich JF, Bach C, Shooshtarizadeh P, Picaud S, Bourcier T, Speeg-Schatz C, Strub JM, Van Dorsselaer A, Corti A, Aunis D, Metz-Boutigue MH.Processing of chromogranins/secretogranin in patients with diabetic retinopathy.Regul Pept. 2011 Feb 25;167(1):118-24.Bougrid A, Claudepierre T, Picaud S, Ayad G, Mornet D, Dorbani-Mamine L, Rendon A, Darbeida H.Expression of Dystrophins and the Dystrophin-Associated-Protein Complex by Pituicytes in Culture.Neurochem Res. 2011 Apr 12. [Epub ahead of print]
Ibad RT, Rheey J, Mrejen S, Forster V, Picaud S, Prochiantz A, Moya KL.Otx2 Promotes the Survival of Damaged Adult Retinal Ganglion Cells and Protects against Excitotoxic Loss of Visual Acuity In Vivo.J Neurosci. 2011 Apr 6;31(14):5495-5503.
Kasi H, Bertsch A, Guyomard JL, Kolomiets B, Picaud S, Pelizzone M, Renaud P.Simulations to study spatial extent of stimulation and effect of electrode-tissue gap in subretinal implants.Med Eng Phys. 2011 Feb 25. [Epub ahead of print]
Fournier I, Gaucher D, Chich JF, Bach C, Shooshtarizadeh P, Picaud S, Bourcier T, Speeg-Schatz C, Strub JM, Van Dorsselaer A, Corti A, Aunis D, Metz-Boutigue MH.Processing of chromogranins/secretogranin in patients with diabetic retinopathy.Regul Pept. 2011 Feb 25;167(1):118-24.
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