direct-coupled-electroretinogram-dc-erg-for-recording-light-evoked

Direct-Coupled Electroretinogram (DC-ERG) for Recording the Light-Evoked Electrical Responses of the Mouse Retinal Pigment Epithelium

The retinal pigment epithelium (RPE) is a specialized monolayer of cells strategically located between the retina and the choriocapillaris that maintain ...

National Institutes of Health

&#xC5F0;&#xB839; &#xAD00;&#xB828; &#xD669; &#xBC18; &#xC138;&#xB300; (AMD)&#xC758; pathogenesis &#xBC0F; &#xBCF4;&#xD638;&#xC790; &#xB2E8;&#xBC31;&#xC9C8;&#xACFC; &#xC2E0;&#xACBD; &#xD1F4;&#xD589; &#xC131; &#xC9C8;&#xD658; &#xC0AC;&#xC774; &#xADF8;&#xB824;&#xC9C4; &#xB9C1;&#xD06C;&#xC5D0;&#xC11C; &#xB9DD;&#xB9C9; &#xC0C9;&#xC18C; &#xC0C1;&#xD53C; (RPE)&#xC758; &#xC911;&#xC694; &#xD55C; &#xC5ED;&#xD560;&#xC744; &#xAC10;&#xC548;&#xD560; &#xB54C;, &#xC6B0;&#xB9AC; AMD&#xC758; Ccl2(-/-)/Cx3cr1(-/-) &#xB9C8;&#xC6B0;&#xC2A4; &#xBAA8;&#xB378;&#xC5D0;&#xC11C; &#xBB38;&#xD654; RPE &#xBAA9;&#xC801;&#xC744; &#xBCF4;&#xD638;&#xC790; &#xB2E8;&#xBC31;&#xC9C8; Erp29&#xC758; &#xC2DD;&#xC744; &#xACC4;&#xC0B0; &#xD569;&#xB2C8;&#xB2E4;.

&#xAD50;&#xC591;&#xB41C; &#xB9DD;&#xB9C9;&#xC5D0; ERp29 &#xC2DD;&#xC758; &#xBE44;&#xADF9;&#xC801; &#xC548;&#xB8CC; &#xC0C1;&#xD53C; &#xC138;&#xD3EC; &#xB098;&#xC774; &#xAD00;&#xB828; &#xD669; &#xBC18; &#xBCC0;&#xC131;&#xC758; Ccl2/Cx3cr1 &#xACB0;&#xD54D; &#xB9C8;&#xC6B0;&#xC2A4; &#xBAA8;&#xB378;&#xC5D0;&#xC11C;.

Dado el papel cr&iacute;tico del epitelio pigmentario de la retina (EPR) en la patogenesia de generaci&oacute;n macular relacionada con la edad (AMD) y los enlaces entre las prote&iacute;nas chaperonas y enfermedades neurodegenerativas, pretende la cultura RPE por el modelo de rat&oacute;n de Ccl2(-/-)/Cx3cr1(-/-) de AMD y evaluar la expresi&oacute;n de la Prote&iacute;na chaperona ERp29.

Constancia de ERp29 expresi&oacute;n en retina cultivada pigmento c&eacute;lulas epiteliales en el modelo de rat&oacute;n deficiente de Ccl2/Cx3cr1 de degeneraci&oacute;n macular relacionada con la edad.

&#x7DB2;&#x819C;&#x8272;&#x7D20;&#x4E0A;&#x76AE; (RPE) &#x306E;&#x91CD;&#x8981;&#x306A;&#x5F79;&#x5272;&#x306E;&#x5E74;&#x9F62;&#x95A2;&#x9023;&#x9EC4;&#x6591;&#x4E16;&#x4EE3; (AMD) &#x306E;&#x75C5;&#x56E0;&#x3068;&#x30B7;&#x30E3;&#x30DA;&#x30ED;&#x30F3;&#x86CB;&#x767D;&#x8CEA;&#x3068;&#x795E;&#x7D4C;&#x5909;&#x6027;&#x75BE;&#x60A3;&#x306E;&#x9593;&#x306B;&#x63CF;&#x304B;&#x308C;&#x305F;&#x30EA;&#x30F3;&#x30AF;&#x3092;&#x4E0E;&#x3048;&#x3001;&#x6211;&#x3005; &#x306F; AMD &#x306E; Ccl2(-/-)/Cx3cr1(-/-) &#x30DE;&#x30A6;&#x30B9; &#x30E2;&#x30C7;&#x30EB;&#x304B;&#x3089;&#x6587;&#x5316; RPE &#x3092;&#x76EE;&#x7684;&#x3057;&#x3001;&#x30B7;&#x30E3;&#x30DA;&#x30ED;&#x30F3;&#x86CB;&#x767D;&#x8CEA; ERp29 &#x306E;&#x5F0F;&#x3092;&#x8A55;&#x4FA1;&#x3057;&#x307E;&#x3059;&#x3002;

ERp29 &#x5F0F;&#x57F9;&#x990A;&#x7DB2;&#x819C;&#x306E;&#x6052;&#x5E38;&#x6027;&#x8272;&#x7D20;&#x4E0A;&#x76AE;&#x7D30;&#x80DE;&#x306E;&#x5E74;&#x9F62;&#x95A2;&#x9023;&#x9EC4;&#x6591;&#x5909;&#x6027;&#x75C7; Ccl2/Cx3cr1 &#x6B20;&#x640D;&#x30DE;&#x30A6;&#x30B9; &#x30E2;&#x30C7;&#x30EB;&#x306E;&#x3002;

&#x6211;&#x4EEC;&#x5728;&#x4E0E;&#x5E74;&#x9F84;&#x6709;&#x5173;&#x7684;&#x9EC4;&#x6591;&#x4EE3; (AMD) &#x7684;&#x53D1;&#x75C5;&#x673A;&#x5236;&#x548C;&#x7ED8;&#x5236;&#x4F34;&#x4FA3;&#x86CB;&#x767D;&#x4E0E;&#x795E;&#x7ECF;&#x9000;&#x884C;&#x6027;&#x75BE;&#x75C5;&#x4E4B;&#x95F4;&#x7684;&#x8054;&#x7CFB;&#x7531;&#x4E8E;&#x89C6;&#x7F51;&#x819C;&#x8272;&#x7D20;&#x4E0A;&#x76AE; (RPE) &#x7684;&#x5173;&#x952E;&#x4F5C;&#x7528;&#xFF0C;&#x4ECE; Ccl2(-/-)/Cx3cr1(-/-) &#x5C0F;&#x9F20;&#x6A21;&#x578B;&#x7684; AMD &#x65E8;&#x5728;&#x57F9;&#x517B;&#x89C6;&#x7F51;&#x819C;&#x8272;&#x7D20;&#x4E0A;&#x76AE;&#x548C;&#x8BC4;&#x4F30;&#x4F34;&#x4FA3;&#x86CB;&#x767D; ERp29 &#x7684;&#x8868;&#x8FBE;&#x3002;

&#x5728;&#x57F9;&#x517B;&#x7684;&#x89C6;&#x7F51;&#x819C;&#x4E2D;&#x7684; ERp29 &#x8868;&#x8FBE;&#x7684;&#x6052;&#x5E38;&#x6027;&#x8272;&#x7D20;&#x4E0A;&#x76AE;&#x7EC6;&#x80DE;&#x7684;&#x5E74;&#x9F84;&#x76F8;&#x5173;&#x6027;&#x9EC4;&#x6591;&#x53D8;&#x6027; Ccl2/Cx3cr1 &#x7F3A;&#x4E4F;&#x5C0F;&#x9F20;&#x6A21;&#x578B;&#x4E2D;&#x3002;

Given the critical role of the retinal pigment epithelium (RPE) in the pathogenesis of age-related macular generation (AMD) and the links drawn between chaperone proteins and neurodegenerative disease, we aimed to culture RPE from the Ccl2(-/-)/Cx3cr1(-/-) mouse model of AMD and evaluate expression of chaperone protein ERp29.

Constancy of ERp29 expression in cultured retinal pigment epithelial cells in the Ccl2/Cx3cr1 deficient mouse model of age-related macular degeneration.

&Eacute;tant donn&eacute; le r&ocirc;le crucial de l&#39;&eacute;pith&eacute;lium pigmentaire r&eacute;tinien (RPE) dans la pathogen&egrave;se de la g&eacute;n&eacute;ration de maculaire li&eacute;e &agrave; l&#39;&acirc;ge (DMLA) et les liens entre les prot&eacute;ines chaperons et maladies neuro-d&eacute;g&eacute;n&eacute;ratives, nous destin&eacute; &agrave; la culture RPE du mod&egrave;le de souris Ccl2(-/-)/Cx3cr1(-/-) de la DMLA et d&#39;&eacute;valuer l&#39;expression de la prot&eacute;ine chaperon ERp29.

Constance de l&#39;expression de ERp29 dans la r&eacute;tine cultiv&eacute;e des pigments des cellules &eacute;pith&eacute;liales dans le mod&egrave;le de souris d&eacute;ficientes de Ccl2/Cx3cr1 de la d&eacute;g&eacute;n&eacute;rescence maculaire li&eacute;e &agrave; l&#39;&acirc;ge.

Angesichts der kritische Rolle des retinalen Pigmentepithels (RPE) in der Pathogenese der altersbedingten Makula-Generation (AMD) und die Verbindungen zwischen Chaperon-Proteine und Neurodegenerative Krankheit, wir sollen Kultur RPE aus dem Ccl2(-/-)/Cx3cr1(-/-)-Maus-Modell von AMD und Ausdruck von Chaperon-Protein ERp29 auswerten.

Konstanz der ERp29 Ausdruck in kultivierten Retinal pigment Epithelzellen im Ccl2/Cx3cr1 mangelhaft Mausmodell der altersbedingten Makula-Degeneration.

Dato il ruolo critico dell&#39;epitelio pigmentato retinico (RPE) nella patogenesi della generazione maculare legata all&#39;et&agrave; (AMD) e i legami tracciati proteine chaperone e malattie neurodegenerative, abbiamo rivolto alla cultura RPE dal modello del mouse Ccl2(-/-)/Cx3cr1(-/-) di AMD e valutare l&#39;espressione della proteina chaperone ERp29.

Costanza di ERp29 espressione colta della retina pigmento cellule epiteliali nel modello del mouse carenti Ccl2/Cx3cr1 della degenerazione maculare legata all&#39;et&agrave;.

: Induced pluripotent stem cells (iPSCs) can be efficiently differentiated into retinal pigment epithelium (RPE), offering the possibility of autologous cell replacement therapy for retinal degeneration stemming from RPE loss. The generation and maintenance of epithelial apical-basolateral polarity is fundamental for iPSC-derived RPE (iPSC-RPE) to recapitulate native RPE structure and function. Presently, no criteria have been established to determine clonal or donor based heterogeneity in the polarization and maturation state of iPSC-RPE. We provide an unbiased structural, molecular, and physiological evaluation of 15 iPSC-RPE that have been derived from distinct tissues from several different donors. We assessed the intact RPE monolayer in terms of an ATP-dependent signaling pathway that drives critical aspects of RPE function, including calcium and electrophysiological responses, as well as steady-state fluid transport. These responses have key in vivo counterparts that together help determine the homeostasis of the distal retina. We characterized the donor and clonal variation and found that iPSC-RPE function was more significantly affected by the genetic differences between different donors than the epigenetic differences associated with different starting tissues. This study provides a reference dataset to authenticate genetically diverse iPSC-RPE derived for clinical applications.

In Pursuit of Authenticity: Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium for Clinical Applications.

MicroRNA-204 (miR-204) is expressed in pulmonary, renal, mammary and eye tissue, and its reduction can result in multiple diseases including cancer. We first generated miR-204-/- mice to study the impact of miR-204 loss on retinal and retinal pigment epithelium (RPE) structure and function. The RPE is fundamentally important for maintaining the health and integrity of the retinal photoreceptors. miR-204-/- eyes evidenced areas of hyper-autofluorescence and defective photoreceptor digestion, along with increased microglia migration to the RPE. Migratory Iba1+ microglial cells were localized to the RPE apical surface where they participated in the phagocytosis of photoreceptor outer segments (POSs) and contributed to a persistent build-up of rhodopsin. These structural, molecular and cellular outcomes were accompanied by decreased light-evoked electrical responses from the retina and RPE. In parallel experiments, we suppressed miR-204 expression in primary cultures of human RPE using anti-miR-204. In vitro suppression of miR-204 in human RPE similarly showed abnormal POS clearance and altered expression of autophagy-related proteins and Rab22a, a regulator of endosome maturation. Together, these in vitro and in vivo experiments suggest that the normally high levels of miR-204 in RPE can mitigate disease onset by preventing generation of oxidative stress and inflammation originating from intracellular accumulation of undigested photoreactive POS lipids. More generally, these results implicate RPE miR-204-mediated regulation of autophagy and endolysosomal interaction as a critical determinant of normal RPE/retina structure and function.

Regulation of phagolysosomal activity by miR-204 critically influences structure and function of retinal pigment epithelium/retina.

Aberrant microRNA (miRNA) expression affects biologic processes and downstream genes that are crucial to CKD initiation or progression. The miRNA miR-204-5p is highly expressed in the kidney but whether miR-204-5p plays any role in the development of chronic renal injury is unknown.

Congxiao Zhang

Ocular and Stem Cell Translational Research Section,
National Eye Institute,
Ocular and Stem Cell Translational Research Section, National Eye Institute

National Institutes of Health

Direct-Coupled Electroretinogram (DC-ERG) for Recording the Light-Evoked Electrical Responses of the Mouse Retinal Pigment Epithelium

Congxiao Zhang

Ocular and Stem Cell Translational Research Section,National Eye Institute,Ocular and Stem Cell Translational Research Section, National Eye Institute

National Institutes of Health

Direct-Coupled Electroretinogram (DC-ERG) for Recording the Light-Evoked Electrical Responses of the Mouse Retinal Pigment Epithelium

Ocular and Stem Cell Translational Research Section,
National Eye Institute,
Ocular and Stem Cell Translational Research Section, National Eye Institute