optimized-protocol-for-retinal-wholemount-preparation-for-imaging

Optimized Protocol for Retinal Wholemount Preparation for Imaging and Immunohistochemistry

Working with delicate tissue can be a complicating factor when performing immunohistochemical assessment. Here, we present a method that utilizes a ...

Weill Medical College of Cornell University

To map the spatiotemporal decline in retinally driven activity in the superior colliculus (SC) of transgenic S334ter-line-3 rats that express a mutated rhodopsin, which causes photoreceptor degeneration. To determine whether transplantation of fetal retinal sheets into the subretinal space of these rats can recover visual activity in the SC.

Retinal transplantation-induced recovery of retinotectal visual function in a rodent model of retinitis pigmentosa.

Inhibition at bipolar cell axon terminals regulates excitatory signaling to ganglion cells and is mediated, in part, by GABAC receptors. We investigated GABAC receptor-mediated inhibition using pharmacological approaches and genetically altered mice that lack GABAC receptors. Responses to applied GABA showed distinct time courses in various bipolar cell classes, attributable to different proportions of GABAA and GABAC receptors. The elimination of GABAC receptors in GABAC null mice reduced and shortened GABA-activated currents and light-evoked inhibitory synaptic currents (L-IPSCs) in rod bipolar cells. ERG measurements and recordings from the optic nerve showed that inner retinal function was altered in GABAC null mice. These data suggest that GABAC receptors determine the time course and extent of inhibition at bipolar cell terminals that, in turn, modulates the magnitude of excitatory transmission from bipolar cells to ganglion cells.

GABAC receptor-mediated inhibition in the retina.

The electroretinogram (ERG) is routinely used to study retinal physiology in the clinic and in research. Due to their outstanding properties, contact lens electrodes (CLEs) are widely used for ERG recordings. Though the procedures for ERG recordings in mice are similar to those used in humans and larger vertebrates, use of CLEs in the mouse has been limited because of difficulties involved with the manufacturing of small contact lenses. We describe a simple instrument and method for manufacturing contact lenses and CLEs for stable ERG recordings in mice. The instrument operates like a hole-punch and is based on slip joint pliers incorporating a ball bearing on one jaw and forming plate on the other. These CLEs are simple to manufacture, inexpensive and provide stable, long-term recordings of corneal ERGs in mice. With minor modifications, these CLEs could be made for other small animals such as rats or fish.

Improved contact lens electrode for corneal ERG recordings in mice.

The aim of this study was to establish synapses between a transplant and a degenerated retina. To tackle this difficult task, a little-known but well-established CNS method was chosen: trans-synaptic pseudorabies virus (PRV) tracing. Sheets of E19 rat retina with or without retinal pigment epithelium (RPE) were transplanted to the subretinal space in 33 Royal College of Surgeons (RCS) and transgenic s334ter-5 rats with retinal degeneration. Several months later, PRV-BaBlu (expressing E. colibeta-galactosidase) or PRV-Bartha was injected into an area of the exposed superior colliculus (SC), topographically corresponding to the transplant placement in the retina. Twenty normal rats served as controls. After survival times of 1-5 days, retinas were examined for virus by X-gal histochemistry, immunohistochemistry and electron microscopy. In normal controls, virus was first seen in retinal ganglion cells and Müller glia after 1-1.5 days, and had spread to all retinal layers after 2-3 days. Virus-labeled cells were found in 16 of 19 transplants where the virus injection had retrogradely labeled the topographically correct transplant area of the host retina. Electron microscopically, enveloped and nonenveloped virus could clearly be detected in infected cells. Enveloped virus was found only in neurons. Infected glial cells contained only nonenveloped virus. Neurons in retinal transplants are labeled after PRV injection into the host brain, indicating synaptic connectivity between transplants and degenerated host retinas. This study provides evidence that PRV spreads in the retina as in other parts of the CNS and is useful to outline transplant-host circuitry.

Transsynaptic virus tracing from host brain to subretinal transplants.

The receptive field (RF) of most retinal ganglion cells (RGCs) is comprised of an excitatory center and an antagonistic surround. Interactions between these RF elements shape most of the visual responses of RGCs. To begin to investigate center-surround interactions of mouse RGCs quantitatively, we characterized their responses in an in vivo preparation to a variety of spot and full-field stimuli. When RGCs were stimulated with a spot that matched the cell's RF center diameter (optimal spot), all RGCs could be categorized as either ON- or OFF-center. In all RGCs, full-field stimulation significantly reduced both the peak and the mean firing rates evoked with an optimal spot stimulus. Full-field stimulation revealed differences in other response properties between ON- and OFF-center RGCs. With a full-field stimulus, the duration of the OFF-center RGCs response was reduced making them more transient, while the duration of the ON-center RGCs increased making them more sustained. Of most interest, full-field stimulation altered the RF center response sign in approximately half of the OFF-center RGCs, which became either OFF/ON or ON only. In contrast, all ON-center and the other OFF-center cells conserved their RF response sign in the presence of the full-field stimulus. We propose that sign-altering OFF-center RGCs possess an additional RF surround mechanism that underlies this alteration in their response. Of general interest these results suggest that the sole use of full-field stimulation to categorize visual response properties of RGCs does not adequately reflect their RF organization and, therefore, is not an optimal strategy for their classification.

Stimulus size and intensity alter fundamental receptive-field properties of mouse retinal ganglion cells in vivo.

Axon terminals from the two eyes initially overlap in the dorsal-lateral geniculate nucleus (dLGN) but subsequently refine to occupy nonoverlapping territories. Retinal activity is required to establish and maintain this segregation. We show that despite the presence of retinal activity, segregated projections desegregate when the structure of activity is altered. Early in development, spontaneous retinal activity in the no b-wave (nob) mouse is indistinguishable from that of wild-type mice, and eye-specific segregation proceeds normally. But, around eye-opening, spontaneous and visually evoked activity in nob retinas become abnormal, coincident with a failure to preserve precise eye-specific territories. Dark-rearing studies suggest that altered visual experience is not responsible. Transgenic rescue of the mutated protein (nyctalopin) within nob retinal interneurons, without rescuing expression in either retinal projection neurons or their postsynaptic targets in the dLGN, restores spontaneous retinal activity patterns and prevents desegregation. Thus, normally structured spontaneous retinal activity stabilizes newly refined retinogeniculate circuitry.

Failure to maintain eye-specific segregation in nob, a mutant with abnormally patterned retinal activity.

Sensory information is thought to be modulated by presynaptic inhibition. Although this form of inhibition is a well-studied phenomenon, it is still unclear what role it plays in shaping sensory signals in intact circuits. By visually stimulating the retinas of transgenic mice lacking GABAc receptor-mediated presynaptic inhibition, we found that this inhibition regulated the dynamic range of ganglion cell (GC) output to the brain. Presynaptic inhibition acted differentially upon two major retinal pathways; its elimination affected GC responses to increments, but not decrements, in light intensity across the visual scene. The GC dynamic response ranges were different because presynaptic inhibition limited glutamate release from ON, but not OFF, bipolar cells, which modulate the extent of glutamate spillover and activation of perisynaptic NMDA receptors at ON GCs. Our results establish a role for presynaptic inhibitory control of spillover in determining sensory output in the CNS.

Presynaptic inhibition modulates spillover, creating distinct dynamic response ranges of sensory output.

Subretinal prosthetics are designed to electrically stimulate second-order cells, replacing dysfunctional photoreceptors in diseases such as retinitis pigmentosa (RP). For functional vision to occur, this signal must also reach central visual structures. In the current study, a subretinally implanted prosthetic was evaluated in the Royal College of Surgeons (RCS) rat model of RP, to determine its capacity to activate the retinotectal pathway.

Stimulation via a subretinally placed prosthetic elicits central activity and induces a trophic effect on visual responses.

The prion protein PrP(C) is infamous for its role in disease, but its normal physiological function remains unknown. Here we found a previously unknown behavioral phenotype of Prnp(-/-) mice in an odor-guided task. This phenotype was manifest in three Prnp knockout lines on different genetic backgrounds, which provides strong evidence that the phenotype is caused by a lack of PrP(C) rather than by other genetic factors. Prnp(-/-) mice also showed altered behavior in a second olfactory task, suggesting that the phenotype is olfactory specific. Furthermore, PrP(C) deficiency affected oscillatory activity in the deep layers of the main olfactory bulb, as well as dendrodendritic synaptic transmission between olfactory bulb granule and mitral cells. Notably, both the behavioral and electrophysiological alterations found in Prnp(-/-) mice were rescued by transgenic neuronal-specific expression of PrP(C). These data suggest that PrP(C) is important in the normal processing of sensory information by the olfactory system.

Olfactory behavior and physiology are disrupted in prion protein knockout mice.

&#x670A;&#x86CB;&#x767D; PrP(C) &#x662F;&#x81ED;&#x540D;&#x662D;&#x8457;&#x7684;&#x75BE;&#x75C5;&#xFF0C;&#x5176;&#x4F5C;&#x7528;&#xFF0C;&#x4F46;&#x5176;&#x6B63;&#x5E38;&#x7684;&#x751F;&#x7406;&#x529F;&#x80FD;&#x4ECD;&#x662F;&#x672A;&#x77E5;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x53D1;&#x73B0;&#x4EE5;&#x524D;&#x672A;&#x77E5;&#x7684;&#x884C;&#x4E3A;&#x8868;&#x578B;&#x7684; Prnp(-/-) &#x5C0F;&#x9F20;&#x5728;&#x6C14;&#x5473;&#x5F15;&#x5BFC;&#x7684;&#x4EFB;&#x52A1;&#x3002;&#x6B64;&#x8868;&#x578B;&#x662F;&#x5BF9;&#x4E0D;&#x540C;&#x9057;&#x4F20;&#x80CC;&#x666F;&#xFF0C;&#x4E09;&#x4E2A;&#x670A;&#x75C5;&#x6BD2;&#x6316;&#x7A7A;&#x884C;&#x4E2D;&#x7684;&#x6E05;&#x5355;&#xFF0C;&#x5176;&#x4E2D;&#x63D0;&#x4F9B;&#x4E86;&#x5F3A;&#x6709;&#x529B;&#x7684;&#x8BC1;&#x636E;&#x8BC1;&#x660E;&#x8868;&#x578B;&#x5BFC;&#x81F4;&#x7684; PrP(C) &#x7684;&#x7F3A;&#x4E4F;&#xFF0C;&#x800C;&#x4E0D;&#x662F;&#x7531;&#x5176;&#x4ED6;&#x9057;&#x4F20;&#x56E0;&#x7D20;&#x3002;Prnp(-/-) &#x5C0F;&#x9F20;&#x4E5F;&#x663E;&#x793A;&#x7B2C;&#x4E8C;&#x55C5;&#x4EFB;&#x52A1;&#xFF0C;&#x8868;&#x660E;&#x55C5;&#x89C9;&#x7684;&#x7279;&#x5B9A;&#x7684;&#x8868;&#x578B;&#x6539;&#x53D8;&#x7684;&#x884C;&#x4E3A;&#x3002;&#x6B64;&#x5916;&#xFF0C;PrP(C) &#x7F3A;&#x5F71;&#x54CD;&#x632F;&#x8361;&#x6D3B;&#x52A8;&#x4E2D;&#x4E3B;&#x8981;&#x7684;&#x55C5;&#x7403;&#xFF0C;&#x4EE5;&#x53CA; dendrodendritic &#x55C5;&#x7403;&#x9897;&#x7C92;&#x548C;&#x50E7;&#x5E3D;&#x7EC6;&#x80DE;&#x4E4B;&#x95F4;&#x7684;&#x7A81;&#x89E6;&#x4F20;&#x9012;&#x7684;&#x6DF1;&#x5C42;&#x3002;&#x503C;&#x5F97;&#x6CE8;&#x610F;&#x7684;&#x662F;&#xFF0C;&#x7531; PrP(C) &#x8F6C;&#x57FA;&#x56E0;&#x795E;&#x7ECF;&#x5143;&#x7279;&#x5F02;&#x6027;&#x8868;&#x8FBE;&#x6551;&#x8D77;&#x53D1;&#x73B0; Prnp(-/-) &#x5C0F;&#x9F20;&#x7684;&#x884C;&#x4E3A;&#x53CA;&#x7535;&#x751F;&#x7406;&#x6539;&#x53D8;&#x3002;&#x8FD9;&#x4E9B;&#x6570;&#x636E;&#x8868;&#x660E; PrP(C) &#x662F;&#x5728;&#x611F;&#x5B98;&#x4FE1;&#x606F;&#x7531;&#x55C5;&#x89C9;&#x7CFB;&#x7EDF;&#x6B63;&#x5E38;&#x5904;&#x7406;&#x7684;&#x91CD;&#x8981;&#x3002;

&#x55C5;&#x7684;&#x884C;&#x4E3A;&#x548C;&#x751F;&#x7406;&#x670A;&#x86CB;&#x767D;&#x57FA;&#x56E0;&#x6572;&#x9664;&#x5C0F;&#x9F20;&#x5728;&#x4E2D;&#x65AD;&#x3002;

La prot&eacute;ine prion PrP(C) est tristement c&eacute;l&egrave;bre pour son r&ocirc;le dans la maladie, mais sa fonction physiologique normale reste inconnue. Ici, nous avons trouv&eacute; un ph&eacute;notype comportemental jusque-l&agrave; inconnu des souris Prnp(-/-) dans une t&acirc;che guid&eacute;e par l&#39;odeur. Ce ph&eacute;notype est manifeste dans trois lignes de knock-out du g&egrave;ne Prnp sur diff&eacute;rents fonds g&eacute;n&eacute;tiques, qui apporte la preuve manifeste que le ph&eacute;notype est caus&eacute; par un manque de PrP(C), plut&ocirc;t que par d&#39;autres facteurs g&eacute;n&eacute;tiques. PRNP(-/-) souris a &eacute;galement montrent des comportement alt&eacute;r&eacute; dans une t&acirc;che deuxi&egrave;me olfactive, ce qui sugg&egrave;re que le ph&eacute;notype est sp&eacute;cifique olfactif. En outre, carence en PrP(C) touch&eacute;s activit&eacute; oscillatoire dans les couches profondes de la bulbe olfactif, ainsi que dendrodendritic la transmission synaptique entre bulbe olfactif granule et les cellules mitrales. Notamment, les deux les alt&eacute;rations comportementales et &eacute;lectrophysiologiques trouv&eacute;es chez la souris Prnp(-/-) ont &eacute;t&eacute; secourues par l&#39;expression neuronale sp&eacute;cifique transg&eacute;nique de PrP(C). Ces donn&eacute;es sugg&egrave;rent que la PrP(C) est importante dans le traitement normal de l&#39;information sensorielle par le syst&egrave;me olfactif.

Physiologie et comportement olfactif sont perturb&eacute;es dans chez des souris knockout prot&eacute;ine prion.

Das Prionprotein PrP(C) ist ber&uuml;chtigt f&uuml;r seine Rolle in der Krankheit, aber seine normale physiologische Funktion ist noch unbekannt. Hier fanden wir einen bisher unbekannten Verhalten Ph&auml;notyp Prnp(-/-) M&auml;use in einem Geruch-gef&uuml;hrte ausf&uuml;hren. Dieser Ph&auml;notyp war Manifest in drei Prnp-k.o.-Linien auf unterschiedlichen genetischen Hintergr&uuml;nden, die starke nachweist, die den Ph&auml;notyp durch einen Mangel an PrP(C) und nicht durch andere genetischen Faktoren verursacht wird. PRNP(-/-) M&auml;usen zeigte auch ge&auml;nderte Verhalten in einer zweiten olfaktorischen Aufgabe, was darauf hindeutet, dass der Ph&auml;notyp olfaktorischen bezogen ist. PrP(C)-Mangel betroffen dar&uuml;ber hinaus Oszillatorische Aktivit&auml;t in die tieferen Schichten der wichtigsten Riechkolben sowie Dendrodendritic synaptischen &Uuml;bertragung zwischen Riechkolben Granulat- und mitral Zellen. Insbesondere wurden die Verhaltens- und elektrophysiologischen Ver&auml;nderungen, die Prnp(-/-) fanden von transgenen neuronal-spezifischen Ausdruck der PrP(C) gerettet. Diese Daten legen nahe, dass PrP(C) in die normale Verarbeitung von sensorischen Informationen von olfaktorischen Systems wichtig.

Olfaktorische Verhalten und Physiologie werden im Prion-Protein-Knockout-M&auml;use gest&ouml;rt.

La proteina prionica PrP(C) &egrave; famoso per il suo ruolo nella malattia, ma la normale funzione fisiologica rimane sconosciuto. Qui abbiamo trovato un fenotipo comportamentale precedentemente sconosciuto di Prnp(-/-) topi in un&#39;attivit&agrave; guidate di odore. Questo fenotipo &egrave; stato manifesto in tre linee di Prnp knockout su diversi background genetico, che fornisce la forte evidenza che il fenotipo &egrave; causato da una mancanza di PrP(C), piuttosto che da altri fattori genetici. PRNP(-/-) topi ha mostrato anche comportamento alterato in un compito secondo olfattivo, suggerendo che il fenotipo &egrave; olfattivo specifico. Inoltre, PrP(C) deficit interessato oscillatorio attivit&agrave; negli strati profondi del bulbo olfattivo principale, come pure dendrodendritic Trasmissione sinaptica tra granulo bulbo olfattivo e cellule della valvola mitrale. In particolare, sia le alterazioni comportamentali ed elettrofisiologiche trovate in Prnp(-/-) topi furono salvate da transgenici neuronale specifica espressione di PrP(C). Questi dati suggeriscono che la PrP(C) &egrave; importante per la normale elaborazione dell&#39;informazione sensoriale dal sistema olfattivo.

Fisiologia e comportamento olfattiva sono interrotti in topi knockout della proteina di prion.

&#x30D7;&#x30EA;&#x30AA;&#x30F3;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x306B;&#x95A2;&#x3059;&#x308B;&#x8A66;&#x9A13;&#x306F;&#x75BE;&#x60A3;&#x306B;&#x304A;&#x3051;&#x308B;&#x305D;&#x306E;&#x5F79;&#x5272;&#x3067;&#x6709;&#x540D;&#x3067;&#x3059;&#x304C;&#x3001;&#x305D;&#x306E;&#x6B63;&#x5E38;&#x306A;&#x751F;&#x7406;&#x7684;&#x6A5F;&#x80FD;&#x306F;&#x4E0D;&#x660E;&#x306E;&#x307E;&#x307E;&#x3002;&#x3053;&#x3053;&#x3067;&#x81ED;&#x6C17;&#x30AC;&#x30A4;&#x30C9;&#x4ED8;&#x304D;&#x306E;&#x30BF;&#x30B9;&#x30AF;&#x3067;&#x306F;&#x3001;&#x4EE5;&#x524D;&#x306F;&#x672A;&#x77E5;&#x306E;&#x884C;&#x52D5;&#x8868;&#x73FE;&#x578B; Prnp(-/-) &#x30DE;&#x30A6;&#x30B9;&#x3092;&#x767A;&#x898B;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x8868;&#x73FE;&#x578B;&#x8868;&#x73FE;&#x578B;&#x306B;&#x95A2;&#x3059;&#x308B;&#x8A66;&#x9A13;&#x306E;&#x6B20;&#x5982;&#x3067;&#x306F;&#x306A;&#x304F;&#x4ED6;&#x306E;&#x907A;&#x4F1D;&#x7684;&#x8981;&#x56E0;&#x306B;&#x3088;&#x3063;&#x3066;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3055;&#x308C;&#x308B;&#x5F37;&#x3044;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x30DE;&#x30CB;&#x30D5;&#x30A7;&#x30B9;&#x30C8; 3 &#x306E; Prnp &#x306E;&#x30CE;&#x30C3;&#x30AF;&#x30A2;&#x30A6;&#x30C8;&#x884C;&#x7570;&#x306A;&#x308B;&#x907A;&#x4F1D;&#x7684;&#x80CC;&#x666F;&#x3067;&#x3042;&#x3063;&#x305F;&#x3002;Prnp(-/-) &#x30DE;&#x30A6;&#x30B9;&#x306F;&#x307E;&#x305F;&#x8868;&#x73FE;&#x578B;&#x55C5;&#x899A;&#x56FA;&#x6709;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x5506;&#x7B2C;&#x4E8C;&#x55C5;&#x899A;&#x30BF;&#x30B9;&#x30AF;&#x3067;&#x306F;&#x3001;&#x5909;&#x66F4;&#x3055;&#x308C;&#x305F;&#x52D5;&#x4F5C;&#x3092;&#x793A;&#x3057;&#x305F;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x306B;&#x95A2;&#x3059;&#x308B;&#x8A66;&#x9A13;&#x6B20;&#x4E4F;&#x55C5;&#x7403;&#x9846;&#x7C92;&#x3068;&#x50E7;&#x5E3D;&#x7D30;&#x80DE;&#x306E;&#x6A39;&#x72B6;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x4F1D;&#x9054;&#x3068;&#x540C;&#x69D8;&#x3001;&#x4E3B;&#x55C5;&#x7403;&#x306E;&#x6DF1;&#x3044;&#x5C64;&#x306E;&#x632F;&#x52D5;&#x6D3B;&#x52D5;&#x306E;&#x5F71;&#x97FF;&#x3092;&#x53D7;&#x3051;&#x307E;&#x3059;&#x3002;&#x7279;&#x306B;&#x3001;Prnp(-/-) &#x30DE;&#x30A6;&#x30B9;&#x304C;&#x898B;&#x3064;&#x304B;&#x308A;&#x307E;&#x3057;&#x305F;&#x4E21;&#x65B9;&#x884C;&#x52D5;&#x304A;&#x3088;&#x3073;&#x751F;&#x7406;&#x53F7;&#x306B;&#x95A2;&#x3059;&#x308B;&#x8A66;&#x9A13;&#x306E;&#x30C8;&#x30E9;&#x30F3;&#x30B9;&#x30B8;&#x30A7;&#x30CB;&#x30C3;&#x30AF;&#x795E;&#x7D4C;&#x7D30;&#x80DE;&#x7279;&#x7570;&#x7684;&#x767A;&#x73FE;&#x306B;&#x3088;&#x3063;&#x3066;&#x6551;&#x51FA;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x30C7;&#x30FC;&#x30BF;&#x306B;&#x95A2;&#x3059;&#x308B;&#x8A66;&#x9A13;&#x304C;&#x55C5;&#x899A;&#x7CFB;&#x306B;&#x3088;&#x308B;&#x77E5;&#x899A;&#x60C5;&#x5831;&#x306E;&#x901A;&#x5E38;&#x306E;&#x51E6;&#x7406;&#x3067;&#x91CD;&#x8981;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x304A;&#x52E7;&#x3081;&#x3057;&#x307E;&#x3059;&#x3002;

&#x55C5;&#x899A;&#x884C;&#x52D5;&#x30FB;&#x751F;&#x7406;&#x30D7;&#x30EA;&#x30AA;&#x30F3;&#x86CB;&#x767D;&#x8CEA;&#x30CE;&#x30C3;&#x30AF;&#x30A2;&#x30A6;&#x30C8; &#x30DE;&#x30A6;&#x30B9;&#x3067;&#x306F;&#x4E2D;&#x65AD;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;

&#xD504;&#xB9AC;&#xC628; &#xB2E8;&#xBC31;&#xC9C8; Prp(c)&#xB294;, &#xC9C8;&#xBCD1;&#xC5D0;&#xC11C;&#xC758; &#xC5ED;&#xD560;&#xC5D0; &#xB300; &#xD55C; &#xC720;&#xBA85;&#xD55C; &#xADF8;&#xB300;&#xB85C; &#xADF8;&#xAC83;&#xC758; &#xC815;&#xC0C1;&#xC801;&#xC778; &#xC0DD;&#xB9AC; &#xAE30;&#xB2A5;&#xC744; &#xC54C; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30; &#xC6B0;&#xB9AC;&#xB294; &#xB0C4;&#xC0C8; &#xAE30;&#xBC18; &#xC791;&#xC5C5;&#xC5D0; Prnp(-/-) &#xB9C8;&#xC6B0;&#xC2A4;&#xC758; &#xC774;&#xC804;&#xC5D0; &#xC54C; &#xC218; &#xC5C6;&#xB294; &#xD589;&#xB3D9; &#xD45C;&#xD604; &#xD615;&#xC744; &#xBC1C;&#xACAC;. &#xC774; &#xD615; phenotype &#xB2E4;&#xB978; &#xC720;&#xC804;&#xC801; &#xC694;&#xC778; &#xBCF4;&#xB2E4;&#xB294; Prp(c)&#xC758; &#xBD80;&#xC871;&#xC5D0; &#xC758;&#xD574; &#xBC1C;&#xC0DD; &#xD558;&#xB294; &#xAC15;&#xB825;&#xD55C; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xD558;&#xB294; &#xB2E4;&#xB978; &#xC720;&#xC804;&#xC801; &#xBC30;&#xACBD;&#xC5D0; &#xC138; Prnp &#xB179;&#xC544;&#xC6C3; &#xC904;&#xC5D0; &#xB9E4;&#xB2C8;&#xD398;&#xC2A4;&#xD2B8;&#xB294;. Prnp(-/-) &#xB9C8;&#xC6B0;&#xC2A4; &#xB610;&#xD55C; phenotype &#xD6C4; &#xAC01; &#xAD00;&#xB828; &#xC81C;&#xC548; &#xB450; &#xBC88;&#xC9F8; &#xD6C4; &#xAC01; &#xC791;&#xC5C5;&#xC5D0;&#xC11C; &#xBCC0;&#xACBD; &#xB41C; &#xB3D9;&#xC791;&#xC744; &#xBCF4;&#xC600;&#xB2E4;. &#xB610;&#xD55C;, PrP(C) &#xACB0;&#xD54D;&#xC99D; dendrodendritic &#x55C5;&#xACFC; &#xB9BD; &#xBC0F; mitral &#xC138;&#xD3EC; &#xAC04;&#xC758; &#xC2DC; &#xB0C5; &#xC2A4; &#xC804;&#xC1A1; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C; &#xC8FC;&#xC694; &#x55C5;&#xC758; &#xAE4A;&#xC740; &#xCE35;&#xC5D0; &#xC9C4;&#xB3D9; &#xD65C;&#xB3D9;&#xC5D0; &#xC601;&#xD5A5;&#xC744;. &#xD2B9;&#xD788;, &#xB450; &#xD589;&#xB3D9; &#xBC0F; electrophysiological &#xBCC0;&#xACBD; Prnp(-/-) &#xB9C8;&#xC6B0;&#xC2A4;&#xC5D0; Prp(c)&#xC758; &#xC720;&#xC804;&#xC790; &#xBCC0;&#xD615; &#xAD00;&#xB828; &#xC2E0;&#xACBD; &#xC2DD;&#xC73C;&#xB85C; &#xAD6C;&#xCD9C; &#xD588;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xB370;&#xC774;&#xD130; PrP(C) &#xD6C4; &#xAC01; &#xC2DC;&#xC2A4;&#xD15C;&#xC5D0; &#xC758;&#xD574; &#xAC10;&#xAC01; &#xC815;&#xBCF4;&#xC758; &#xC815;&#xC0C1;&#xC801;&#xC778; &#xCC98;&#xB9AC;&#xC5D0; &#xC911;&#xC694; &#xD558;&#xB2E4;&#xB294; &#xAC83;&#xC774; &#xC88B;&#xC2B5;&#xB2C8;&#xB2E4;.

&#xD6C4; &#xAC01; &#xB3D9;&#xC791; &#xBC0F; &#xC0DD;&#xB9AC;&#xD559;&#xC740; &#xD504;&#xB9AC;&#xC628; &#xB2E8;&#xBC31;&#xC9C8; &#xB179;&#xC544;&#xC6C3; &#xC0DD;&#xC950;&#xC5D0;&#xC11C; &#xC911;&#xB2E8; &#xB429;&#xB2C8;&#xB2E4;.

La prote&iacute;na del pri&oacute;n PrP(C) es famoso por su papel en la enfermedad, pero se desconoce su funci&oacute;n fisiol&oacute;gica normal. Aqu&iacute; encontramos un fenotipo conductual previamente desconocido de ratones Prnp(-/-) en una tarea guiados por el olor. Este fenotipo era manifiesto en tres l&iacute;neas de nocaut de Prnp en diferentes fondos gen&eacute;ticos, que proporciona fuertes evidencias de que el fenotipo es causado por una falta de PrP(C) en lugar de otros factores gen&eacute;ticos. PRNP(-/-) ratones tambi&eacute;n mostraron comportamiento alterado en una tarea segundo olfativa, lo que sugiere que el fenotipo es espec&iacute;fica olfativa. Adem&aacute;s, PrP(C) deficiencia afecta la actividad oscilatoria en las capas profundas de la principal bulbo olfatorio, as&iacute; como transmisi&oacute;n sin&aacute;ptica dendrodendritic entre gr&aacute;nulo de bulbo olfatorio y las c&eacute;lulas mitrales. En particular, las alteraciones conductuales y electrofisiol&oacute;gicas encontradas ratones Prnp(-/-) fueron rescatadas por transg&eacute;nico expresi&oacute;n neuronal espec&iacute;fica de PrP(C). Estos datos sugieren que la PrP(C) es importante en el proceso normal de la informaci&oacute;n sensorial por el sistema olfativo.

Fisiolog&iacute;a y comportamiento olfativo se interrumpen en ratones de knockout de la prote&iacute;na del pri&oacute;n.

&#x6210;&#x4EBA;&#x795E;&#x7ECF;&#x518D;&#x751F;&#x808C;&#x80A4;&#x55C5;&#x7403; (OB) interneurons &#x5728;&#x5927;&#x591A;&#x6570;&#x54FA;&#x4E73;&#x52A8;&#x7269;&#x7684;&#x4E00;&#x751F;&#xFF0C;&#x4F46;&#x671F;&#x95F4;&#x6B64;&#x6052;&#x6D41;&#x91CF;&#x4ECD;&#x7136;&#x4E0D;&#x6E05;&#x695A;&#x5982;&#x4F55; OB &#x4FDD;&#x6301;&#x6052;&#x5B9A;&#x7684;&#x7ED3;&#x6784;&#x548C;&#x529F;&#x80FD;&#x3002;OB &#x9F20;&#x6807;&#xFF0C;&#x5728;&#x6211;&#x4EEC;&#x8C03;&#x67E5;&#x8425;&#x4E1A;&#x989D;&#xFF0C;&#x5176;&#x5F71;&#x54CD;&#x55C5;&#x89C9;&#x529F;&#x80FD;&#x7684;&#x52A8;&#x6001;&#x70E7;&#x8680;&#x6210;&#x4EBA;&#x795E;&#x7ECF;&#x53D1;&#x751F;&#x5728; sub-ventricular &#x533A; &#xFF08;SVZ) x &#x7EBF;&#x75C5;&#x53D8;&#x3002;&#x65E0;&#x8BBA;&#x5BF9; SVZ &#x75C5;&#x53D8;&#x7684;&#x4E25;&#x91CD;&#x6027;&#xFF0C;&#x6211;&#x4EEC;&#x53D1;&#x73B0;&#x6CA1;&#x6709;&#x6539;&#x53D8;&#x7EC6;&#x80DE;&#x5B58;&#x6D3B;&#x7684;&#x5E74;&#x8F7B;&#x6210;&#x4EBA;&#x51FA;&#x751F;&#x7684;&#x9897;&#x7C92; (Gc) &#x51FA;&#x751F;&#x540E;&#x75C5;&#x53D8;&#x90E8;&#x4F4D;&#x548C;&#x75C5;&#x53D8;&#x90E8;&#x4F4D;&#x4E4B;&#x524D;&#x51FA;&#x751F;&#x7684;&#x5730;&#x65B9;&#x9009;&#x533A;&#x7684;&#x4EBA;&#x53E3;&#x9010;&#x6E10;&#x51CF;&#x5C11;&#x3002;&#x7ECF;&#x8FC7;&#x751F;&#x4EA7;&#x4F20;&#x5165;&#x6210;&#x4EBA; OB &#x6240;&#x751F;&#x7684; Gc &#x7684;&#x51CF;&#x5C11;&#x4E86; 96%&#x7684;&#x75C5;&#x53D8;&#xFF0C;&#x6211;&#x4EEC;&#x53D1;&#x73B0;&#x51CF;&#x5C11;&#x884C;&#x4E3A;&#x6050;&#x60E7;&#x53CD;&#x5E94;&#x6761;&#x4EF6;&#x6027;&#x7684;&#x6C14;&#x5473;&#x63D0;&#x793A;&#xFF0C;&#x800C;&#x4E0D;&#x662F;&#x58F0;&#x97F3;&#x63D0;&#x793A;&#x3002;&#x6709;&#x8DA3;&#x7684;&#x662F;&#xFF0C;&#x5C3D;&#x7BA1;&#x8FD9;&#x79CD;&#x884C;&#x4E3A;&#x7684;&#x8D64;&#x5B57;&#x548C;&#x9010;&#x6B65;&#x7684;&#x89E3;&#x5256;&#x7ED3;&#x6784;&#x53D8;&#x5316;&#xFF0C;&#x6211;&#x4EEC;&#x53D1;&#x73B0;&#x6CA1;&#x6709;&#x7535;&#x751F;&#x7406;&#x53D8;&#x5316;&#x4E2D;&#x6D4B;&#x5B9A;&#x7684; GC &#x4EBA;&#x53E3;&#x4F53;&#x5185;&#x901A;&#x8FC7;&#x53D1;&#x751F;&#x6811;&#x7A81;&#x7A81;&#x89E6;&#x53EF;&#x5851;&#x6027;&#x548C;&#x6C14;&#x5473;&#x8BF1;&#x53D1;&#x5C40;&#x90E8;&#x573A;&#x7535;&#x4F4D;&#x632F;&#x8361;&#x3002;&#x8FD9;&#x4E9B;&#x6570;&#x636E;&#x8868;&#x660E;&#x9897;&#x7C92;&#x7EC6;&#x80DE;&#x5C42;&#x4E2D;&#x7684;&#x8425;&#x4E1A;&#x989D;&#x4E00;&#x822C;&#x8131;&#x94A9;&#x7387;&#x7684;&#x6210;&#x4EBA;&#x795E;&#x7ECF;&#x53D1;&#x751F;&#xFF0C;&#x5728;&#x8BE5; OB &#x6210;&#x4EBA;&#x795E;&#x7ECF;&#x53D1;&#x751F;&#x4E2D;&#x7684;&#x4F5C;&#x7528;&#x5E7F;&#x6CDB;&#x884C;&#x4E3A;&#x7684;&#x7CFB;&#x7EDF;&#x8D85;&#x51FA;&#x600E;&#x6837;&#xFF1F;

&#x6D88;&#x878D;&#x7684;&#x9F20;&#x6807;&#x6210;&#x4EBA;&#x795E;&#x7ECF;&#x53D1;&#x751F;&#x6539;&#x53D8;&#x7ED3;&#x6784;&#x55C5;&#x7403;&#x548C;&#x55C5;&#x6050;&#x60E7;&#x7A7A;&#x8C03;&#x3002;

Adult neurogenesis replenishes olfactory bulb (OB) interneurons throughout the life of most mammals, yet during this constant flux it remains unclear how the OB maintains a constant structure and function. In the mouse OB, we investigated the dynamics of turnover and its impact on olfactory function by ablating adult neurogenesis with an x-ray lesion to the sub-ventricular zone (SVZ). Regardless of the magnitude of the lesion to the SVZ, we found no change in the survival of young adult born granule cells (GCs) born after the lesion, and a gradual decrease in the population of GCs born before the lesion. After a lesion producing a 96% reduction of incoming adult born GCs to the OB, we found a diminished behavioral fear response to conditioned odor cues but not to audio cues. Interestingly, despite this behavioral deficit and gradual anatomical changes, we found no electrophysiological changes in the GC population assayed in vivo through dendro-dendritic synaptic plasticity and odor-evoked local field potential oscillations. These data indicate that turnover in the granule cell layer is generally decoupled from the rate of adult neurogenesis, and that OB adult neurogenesis plays a role in a wide behavioral system extending beyond the OB.

Ablation of mouse adult neurogenesis alters olfactory bulb structure and olfactory fear conditioning.

Neurogen&egrave;se adulte r&eacute;approvisionne interneurones du bulbe olfactif (OB) tout au long de la vie de la plupart des mammif&egrave;res, mais au cours de ce flux constant, on voit mal comment l&#39;OB maintient une structure constante et une fonction. Chez la souris OB, nous avons &eacute;tudi&eacute; la dynamique du chiffre d&#39;affaires et son impact sur la fonction olfactive par ablation de la neurogen&egrave;se adulte avec une l&eacute;sion de rayons x &agrave; la zone sub-ventricular (SVZ). Quel que soit l&#39;ampleur de la l&eacute;sion &agrave; la SVZ, nous avons ne constat&eacute; aucun changement dans la survie des cellules (GCs) de la granule de n&eacute; adultes jeunes n&eacute;e apr&egrave;s la l&eacute;sion et une diminution progressive de la population de GCs n&eacute;s avant la l&eacute;sion. Apr&egrave;s une l&eacute;sion, produisant une r&eacute;duction de 96 % des entrants adultes n&eacute;s GCs de l&#39;OB, nous avons trouv&eacute; une r&eacute;ponse diminu&eacute;e peur comportementales &agrave; des stimulus conditionn&eacute; odeur mais pas aux signaux audio. Fait int&eacute;ressant, malgr&eacute; ce d&eacute;ficit comportemental et des changements anatomiques progressive, nous avons ne trouv&eacute; aucune modifications &eacute;lectrophysiologiques dans la population de GC dos&eacute;e in vivo par la plasticit&eacute; synaptique dendro-dendritique et oscillations potentiel de champ local induite par l&#39;odeur. Ces donn&eacute;es indiquent que le chiffre d&#39;affaires dans la couche de cellules de granule est g&eacute;n&eacute;ralement d&eacute;coupl&eacute; du taux de la neurogen&egrave;se adulte, et cette neurogen&egrave;se adulte OB joue un r&ocirc;le dans un syst&egrave;me comportemental large s&#39;&eacute;tendant au-del&agrave; de l&#39;ob.

Ablation de la neurogen&egrave;se adulte souris modifie la structure de bulbe olfactif et conditionnement olfactif de la peur.

Adulten Neurogenese f&uuml;llt die Riechkolben (OB) Interneuronen w&auml;hrend der gesamten Lebensdauer der meisten S&auml;ugetiere, noch w&auml;hrend diese st&auml;ndigen Wandel bleibt unklar, wie die OB eine Konstante Struktur und Funktion verwaltet. In der Maus OB untersucht wir die Dynamik von Umsatz und seine Auswirkungen auf Riechverm&ouml;gen durch Abtragen der adulten Neurogenese mit einer R&ouml;ntgen-L&auml;sion der sub-ventricular Zone (SVZ). Unabh&auml;ngig von dem Ausma&szlig; der L&auml;sion in der SVZ fanden wir keine &Auml;nderung in das &Uuml;berleben der jungen Erwachsenen geboren Untermodul Zellen (GCs), geb. nach L&auml;sion und eine allm&auml;hliche Abnahme der Bev&ouml;lkerung der GCs geboren vor der L&auml;sion. Nach einer L&auml;sion, produzieren eine Reduzierung der eingehenden Erwachsener GCs geboren, um die OB 96 % fanden wir eine verminderte Verhaltens Angst Reaktion konditionierten Geruch Hinweise aber nicht audio-Signale. Interessanterweise fand trotz dieses Verhaltens Defizit und schrittweise anatomischen Ver&auml;nderungen, wir keine elektrophysiologischen Ver&auml;nderungen in der GC-Bev&ouml;lkerung normalisiert in-vivo durch Dendro-dendritische synaptische Plastizit&auml;t und Geruch hervorgerufen lokales Feld m&ouml;glicher Schwingungen. Diese Daten zeigen, dass der Umsatz in dem Granulat Zellschicht ist im Allgemeinen von der Rate der adulten Neurogenese entkoppelt, und, OB adulten Neurogenese spielt eine Rolle in einem breiten Verhaltens System, die jenseits der OB.

Ablation der adulten Neurogenese Maus &auml;ndert Riechkolben Struktur und olfaktorischen Furcht-Konditionierung.

Neurogenesi adulta riempie interneurons bulbo olfattivo (OB) per tutta la vita della maggior parte dei mammiferi, ma durante questo flusso costante rimane poco chiaro come l&#39;OB mantiene una struttura costante e la funzione. Nel topo OB, abbiamo studiato la dinamica del fatturato e il suo impatto sulla funzione olfattiva di ablazione neurogenesi adulta con una lesione radiografia alla zona sub-ventricular (SVZ). Indipendentemente dalla grandezza della lesione per il SVZ, abbiamo non trovato nessun cambiamento nella sopravvivenza delle cellule giovani adulti nati granulo (GCs) nata dopo la lesione e una graduale diminuzione della popolazione di GCs nati prima della lesione. Dopo una lesione producendo una riduzione di 96% di adulti in arrivo nato GCs per l&#39;OB, abbiamo trovato una risposta diminuita paura comportamentale condizionata odore spunti ma non segnali audio. &Egrave; interessante notare che, nonostante questo deficit comportamentali e graduali cambiamenti anatomici, abbiamo non trovato nessun cambiamenti elettrofisiologico nella popolazione GC dosata in vivo attraverso la plasticit&agrave; sinaptica fornite-dendritiche e le oscillazioni di potenziale evocato odore campo locale. Questi dati indicano che il fatturato nello strato delle cellule granulo generalmente &egrave; disaccoppiato dal tasso di neurogenesi adulta, e che neurogenesi adulta OB svolge un ruolo in un sistema ampio comportamentale che si estende oltre la OB.

Ablazione della neurogenesi adulta del mouse altera la struttura del bulbo olfattivo e olfattive paura condizionata.

&#x6210;&#x4F53;&#x795E;&#x7D4C;&#x65B0;&#x751F;&#x3092;&#x4E00;&#x751F;&#x306E;&#x307B;&#x3068;&#x3093;&#x3069;&#x306E;&#x54FA;&#x4E73;&#x985E;&#x306E;&#x55C5;&#x7403; (OB) &#x4ECB;&#x5728;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x88DC;&#x5145;&#x307E;&#x3060;&#x3053;&#x306E;&#x4E00;&#x5B9A;&#x306E;&#x30D5;&#x30E9;&#x30C3;&#x30AF;&#x30B9;&#x4E2D;&#x305D;&#x308C; OB &#x304C;&#x4E00;&#x5B9A;&#x306E;&#x69CB;&#x9020;&#x3068;&#x6A5F;&#x80FD;&#x3092;&#x7DAD;&#x6301;&#x3059;&#x308B;&#x65B9;&#x6CD5;&#x4E0D;&#x660E;&#x77AD;&#x306A;&#x307E;&#x307E;&#x3067;&#x3059;&#x3002;OB &#x306E;&#x30DE;&#x30A6;&#x30B9;&#x6210;&#x4F53;&#x795E;&#x7D4C;&#x65B0;&#x751F;&#x3092; x &#x7DDA;&#x75C5;&#x5909;&#x3092; sub-ventricular &#x30BE;&#x30FC;&#x30F3; (SVZ) &#x3092;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3060;&#x58F2;&#x308A;&#x4E0A;&#x3052;&#x9AD8;&#x3068;&#x55C5;&#x899A;&#x6A5F;&#x80FD;&#x3078;&#x306E;&#x5F71;&#x97FF;&#x306E;&#x30C0;&#x30A4;&#x30CA;&#x30DF;&#x30AF;&#x30B9;&#x3092;&#x8ABF;&#x3079;&#x305F;&#x3002;&#x4E0B;&#x5E2F;&#x306B;&#x75C5;&#x5909;&#x306E;&#x5927;&#x304D;&#x3055;&#x306B;&#x95A2;&#x4FC2;&#x306A;&#x304F;&#x3001;&#x6211;&#x3005; &#x306E;&#x5909;&#x66F4;&#x306F;&#x3001;&#x75C5;&#x5909;&#x3068; Gc &#x306E;&#x524D;&#x306B;&#x75C5;&#x5909;&#x3092;&#x751F;&#x307E;&#x308C;&#x306E;&#x4EBA;&#x53E3;&#x306F;&#x3060;&#x3093;&#x3060;&#x3093;&#x6E1B;&#x3063;&#x3066;&#x5F8C;&#x306B;&#x751F;&#x307E;&#x308C;&#x305F;&#x82E5;&#x3044;&#x5927;&#x4EBA;&#x751F;&#x307E;&#x308C;&#x9846;&#x7C92;&#x7D30;&#x80DE; (Gc) &#x306E;&#x751F;&#x5B58;&#x898B;&#x3064;&#x304B;&#x308A;&#x307E;&#x305B;&#x3093;&#x3067;&#x3057;&#x305F;&#x3002;GCs OB &#x306B;&#x751F;&#x307E;&#x308C;&#x7740;&#x4FE1;&#x5927;&#x4EBA;&#x306E; 96% &#x524A;&#x6E1B;&#x3092;&#x751F;&#x7523;&#x3001;&#x75C5;&#x5909;&#x5F8C;&#x6E1B;&#x5C11;&#x306E;&#x884C;&#x52D5;&#x306E;&#x6050;&#x6016;&#x306E;&#x5FDC;&#x7B54;&#x30A8;&#x30A2;&#x30B3;&#x30F3;&#x81ED;&#x624B;&#x304C;&#x304B;&#x308A;&#x304C;&#x30AA;&#x30FC;&#x30C7;&#x30A3;&#x30AA;&#x624B;&#x304C;&#x304B;&#x308A;&#x3092;&#x767A;&#x898B;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x8208;&#x5473;&#x6DF1;&#x3044;&#x3053;&#x3068;&#x306B;&#x3001;&#x3053;&#x306E;&#x884C;&#x52D5;&#x306E;&#x8D64;&#x5B57;&#x3068;&#x6BB5;&#x968E;&#x7684;&#x306A;&#x89E3;&#x5256;&#x5B66;&#x7684;&#x5909;&#x5316;&#x306B;&#x3082;&#x304B;&#x304B;&#x308F;&#x3089;&#x305A;&#x3001;&#x79C1;&#x305F;&#x3061;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x7684;&#x5909;&#x66F4;&#x306E;&#x8A66;&#x91D1;&#x3055;&#x308C;&#x308B; GC &#x306E;&#x4EBA;&#x53E3;&#x306B;&#x30C7;&#x30F3;&#x30C9;&#x30ED;&#x30D3;&#x30E5;&#x30FC;&#x30E0;&#x6A39;&#x72B6;&#x7A81;&#x8D77;&#x306E;&#x30B7;&#x30CA;&#x30D7;&#x30B9;&#x53EF;&#x5851;&#x6027;&#x3068;&#x30ED;&#x30FC;&#x30AB;&#x30EB; &#x30D5;&#x30A3;&#x30FC;&#x30EB;&#x30C9;&#x81ED;&#x6C17;&#x8A98;&#x767A;&#x96FB;&#x4F4D;&#x632F;&#x52D5;&#x3092;&#x751F;&#x4F53;&#x5185;&#x898B;&#x3064;&#x304B;&#x308A;&#x307E;&#x305B;&#x3093;&#x3002;&#x9846;&#x7C92;&#x7D30;&#x80DE;&#x5C64;&#x306E;&#x8EE2;&#x63DB;&#x306F;&#x3001;&#x4E00;&#x822C;&#x7684;&#x306B;&#x6210;&#x4F53;&#x795E;&#x7D4C;&#x65B0;&#x751F;&#x306E;&#x30EC;&#x30FC;&#x30C8;&#x304B;&#x3089;&#x5207;&#x308A;&#x96E2;&#x3055;&#x308C;&#x3001;&#x305D;&#x306E; OB &#x6210;&#x4F53;&#x3067;&#x306F;&#x5F79;&#x5272;&#x3092;&#x679C;&#x305F;&#x3057;&#x3066;&#x3044;&#x308B; OB &#x3092;&#x8D85;&#x3048;&#x3066;&#x62E1;&#x5F35;&#x3059;&#x308B;&#x5E83;&#x3044;&#x884C;&#x52D5;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x3067;&#x3053;&#x308C;&#x3089;&#x306E;&#x30C7;&#x30FC;&#x30BF;&#x3092;&#x793A;&#x3059;

&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306E;&#x30DE;&#x30A6;&#x30B9;&#x6210;&#x4F53;&#x55C5;&#x7403;&#x306E;&#x69CB;&#x9020;&#x3068;&#x55C5;&#x6050;&#x6016;&#x6761;&#x4EF6;&#x3065;&#x3051;&#x3092;&#x5909;&#x66F4;&#x3057;&#x307E;&#x3059;&#x3002;

&#xC131;&#xCCB4; &#xBAA8;&#xC544; &#x55C5; (OB) &#xC218; &#xB300;&#xBD80;&#xBD84;&#xC758; &#xD3EC;&#xC720; &#xB3D9;&#xBB3C;&#xC758; &#xC218;&#xBA85; &#xC804;&#xCCB4;&#xC5D0; &#xAC78;&#xCCD0; &#xC544;&#xC9C1; &#xC77C;&#xC815; &#xC720;&#xB7C9;&#xC774; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xADF8;&#xAC83;&#xC740; &#xBD88;&#xBD84;&#xBA85; &#xB0A8;&#xC544; &#xC0B0;&#xBD80;&#xC778;&#xACFC; &#xC9C0;&#xC18D;&#xC801;&#xC778; &#xAD6C;&#xC870;&#xC640; &#xAE30;&#xB2A5;&#xC744; &#xC720;&#xC9C0; &#xD558;&#xB294; &#xBC29;&#xBC95;. OB &#xB9C8;&#xC6B0;&#xC2A4;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC; ablating sub-ventricular &#xC601;&#xC5ED; (SVZ) x-&#xC120; &#xBCD1; &#xBCC0;&#xC73C;&#xB85C; &#xC131;&#xCCB4; &#xD68C;&#xC804;&#xC728; &#xBC0F; &#xD6C4; &#xAC01; &#xAE30;&#xB2A5;&#xC5D0; &#xBBF8;&#xCE58;&#xB294; &#xC601;&#xD5A5;&#xC758; &#xC5ED;&#xD559; &#xC870;&#xC0AC;. SVZ &#xD558; &#xBCD1; &#xBCC0; &#xD06C;&#xAE30;&#xC5D0; &#xAD00;&#xACC4; &#xC5C6;&#xC774; &#xC6B0;&#xB9AC; &#xBCD1; &#xBCC0;, &#xBCD1; &#xBCC0; &#xC804;&#xC5D0; &#xD0DC;&#xC5B4;&#xB09C; Gc&#xC758; &#xC778;&#xAD6C;&#xC5D0;&#xC11C; &#xC810;&#xC9C4;&#xC801; &#xAC10;&#xC18C; &#xD6C4; &#xD0DC;&#xC5B4;&#xB09C; &#xC80A;&#xC740; &#xC131;&#xC778; &#xD0C4;&#xC0DD;&#xB41C;&#xACFC; &#xB9BD; &#xC138;&#xD3EC; (Gc)&#xC758; &#xC0DD;&#xC874;&#xC5D0; &#xC544;&#xBB34;&#xB7F0; &#xBCC0;&#xD654;&#xB97C; &#xBC1C;&#xACAC;. Gc &#xC0B0;&#xBD80;&#xC778;&#xACFC; &#xD0DC;&#xC5B4;&#xB09C; &#xB4E4;&#xC5B4;&#xC624;&#xB294; &#xC131;&#xC778;&#xC758; 96% &#xAC10;&#xC18C;&#xB97C; &#xC0DD;&#xC0B0; &#xD55C; &#xBCD1; &#xBCC0;, &#xD6C4; &#xC6B0;&#xB9AC;&#xB294; &#xBC14;&#xB978;&#xB41C; &#xB0C4;&#xC0C8; &#xB2E8;&#xC11C; &#xC544;&#xB2C8;&#xB77C; &#xC624;&#xB514;&#xC624; &#xC2E0;&#xD638; &#xC800;&#xD558; &#xD589;&#xB3D9; &#xACF5;&#xD3EC; &#xBC18;&#xC751;&#xC744; &#xBC1C;&#xACAC;. &#xD765;&#xBBF8;&#xB86D;&#xAC8C;&#xB3C4;,&#xC774; &#xD589;&#xB3D9; &#xC801;&#xC790;&#xC640; &#xC810;&#xC9C4;&#xC801; &#xD574;&#xBD80;&#xD559; &#xC801; &#xBCC0;&#xD654;&#xC5D0;&#xB3C4; &#xBD88;&#xAD6C; &#xD558; &#xACE0; &#xC6B0;&#xB9AC; &#xC788;&#xB294; electrophysiological &#xBCC0;&#xACBD; &#xBD84;&#xC11D; GC &#xC778;&#xAD6C; dendro &#xC218;&#xC9C0;&#xC0C1; &#xC2DC; &#xB0C5; &#xC2A4;&#xAC00; &#xC18C;&#xC131;&#xC744; &#xD1B5;&#xD574; &#xB85C;&#xCEEC; &#xD544;&#xB4DC; &#xB0C4;&#xC0C8; &#xAC16;&#xB294; &#xC7A0;&#xC7AC;&#xC801;&#xC778; &#xC9C4;&#xB3D9; vivo&#xC5D0;&#xC11C;. &#xC774;&#xB7EC;&#xD55C; &#xB370;&#xC774;&#xD130;&#xB97C; &#xB098;&#xD0C0;&#xB0B4;&#xB294; &#xD68C;&#xC804;&#xC728;&#xACFC; &#xB9BD; &#xC138;&#xD3EC; &#xCE35;&#xC5D0;&#xB294; &#xC77C;&#xBC18;&#xC801;&#xC73C;&#xB85C; &#xC131;&#xCCB4;, &#xC18D;&#xB3C4;&#xC5D0;&#xC11C; &#xBD84;&#xB9AC; &#xD558; &#xACE0; &#xADF8; &#xC0B0;&#xBD80;&#xC778;&#xACFC; &#xC131;&#xCCB4; &#xC0B0;&#xBD80;&#xC778;&#xACFC; &#xB118;&#xC5B4; &#xD655;&#xC7A5; &#xB113;&#xC740; &#xD589;&#xB3D9; &#xC2DC;&#xC2A4;&#xD15C;&#xC5D0; &#xC5ED;&#xD560;&#xC744; &#xD55C;&#xB2E4;

&#xB9C8;&#xC6B0;&#xC2A4; &#xC131;&#xCCB4;&#xC758; &#xC808;&#xC81C; &#x55C5; &#xAD6C;&#xC870; &#xBC0F; &#xD6C4; &#xAC01; &#xB450;&#xB824;&#xC6C0; &#xCEE8;&#xB514;&#xC154;&#xB2DD; &#xBCC0;&#xACBD;&#xD569;&#xB2C8;&#xB2E4;.

La neurog&eacute;nesis adulta repone interneuronas del bulbo olfatorio (OB) a lo largo de la vida de la mayor&iacute;a de los mam&iacute;feros, sin embargo, durante este flujo constante no es claro c&oacute;mo el OB mantiene una estructura constante y funci&oacute;n. En el rat&oacute;n OB, investigamos la din&aacute;mica de rotaci&oacute;n y su impacto en la funci&oacute;n olfatoria por ablaci&oacute;n neurog&eacute;nesis adulta con una lesi&oacute;n de rayos x a la zona sub-ventricular (SVZ). Independientemente de la magnitud de la lesi&oacute;n a la SVZ, no encontramos ning&uacute;n cambio en la supervivencia de las c&eacute;lulas del gr&aacute;nulo nacido adulto joven (GCs) nacida despu&eacute;s de la lesi&oacute;n y una disminuci&oacute;n gradual de la poblaci&oacute;n de GCs nacidos antes de la lesi&oacute;n. Despu&eacute;s de una lesi&oacute;n que se produce una reducci&oacute;n del 96% de entrante adulto nacido GCs al OB, encontramos una respuesta disminuida comportamiento miedo a se&ntilde;ales de olor condicionada pero no a las se&ntilde;ales de audio. Curiosamente, a pesar de este d&eacute;ficit conductual y graduales cambios anat&oacute;micos, no encontramos cambios electrofisiol&oacute;gicos en la poblaci&oacute;n de GC ensayada en vivo a trav&eacute;s de la plasticidad sin&aacute;ptica dendro-dendr&iacute;tica y oscilaciones potenciales evocados por el olor del campo local. Estos datos indican que el volumen de negocios en la capa de c&eacute;lulas del gr&aacute;nulo es generalmente desacoplado de la tasa de neurog&eacute;nesis adulta, y esa neurog&eacute;nesis adulta OB desempe&ntilde;a un papel en un sistema amplio de comportamiento que se extiende m&aacute;s all&aacute; de la OB.

Ablaci&oacute;n de la neurog&eacute;nesis adulta de rat&oacute;n altera la estructura del bulbo olfatorio y acondicionado de miedo olfativa.

The visual system is highly sensitive to dynamic features in the visual scene. However, it is not known how or where this enhanced sensitivity first occurs. We investigated this phenomenon by studying interactions between excitatory and inhibitory synapses in the second synaptic layer of the mouse retina. We found that these interactions showed activity-dependent changes that enhanced signaling of dynamic stimuli. Excitatory signaling from cone bipolar cells to ganglion cells exhibited strong synaptic depression, attributable to reduced glutamate release from bipolar cells. This depression was relieved by amacrine cell inhibitory feedback that activated presynaptic GABA(C) receptors. We found that the balance between excitation and feedback inhibition depended on stimulus frequency; at short interstimulus intervals, excitation was enhanced, attributable to reduced inhibitory feedback. This dynamic interplay may enrich visual processing by enhancing retinal responses to closely spaced temporal events, representing rapid changes in the visual environment.

Nonlinear interactions between excitatory and inhibitory retinal synapses control visual output.

Neural oscillations play an important role in normal brain activity, but also manifest during Parkinson's disease, epilepsy, and other pathological conditions. The contribution of these aberrant oscillations to the function of the surviving brain remains unclear. In recording from retina in a mouse model of retinal degeneration (RD), we found that the incidence of oscillatory activity varied across different cell classes, evidence that some retinal networks are more affected by functional changes than others. This aberrant activity was driven by an independent inhibitory amacrine cell oscillator. By stimulating the surviving circuitry at different stages of the neurodegenerative process, we found that this dystrophic oscillator further compromises the function of the retina. These data reveal that retinal remodeling can exacerbate the visual deficit, and that aberrant synaptic activity could be targeted for RD treatment.

Network deficiency exacerbates impairment in a mouse model of retinal degeneration.

In this manuscript, we describe a protocol for capturing both physiological and structural properties of living neuronal tissue. An essential aspect of this method is its flexibility and fast turnaround time. It is a streamlined process that includes recording of electrophysiological neuronal activity, calcium imaging, and structural analysis. This is accomplished by placing intact tissue on a modified Millicell Biopore insert. The Biopore membrane suspends the tissue in the perfusion solution, allowing for complete access to nutrients, oxygen, and pharmacological agents. The ring that holds the membrane ensures its structural stability; forceps can be used to grip the ring without contacting the filter or the tissue, for easy transfer between multiple setups. We show that tissue readily adheres to the surface of the membrane, its entire surface is visible in transmitted light and accessible for recording and imaging, and remains responsive to physiological stimuli for extended periods of time.

A time and cost efficient approach to functional and structural assessment of living neuronal tissue.

Retinal degeneration leads to progressive photoreceptor cell death, resulting in vision loss. Subsequently, inner retinal neurons develop aberrant synaptic activity, compounding visual impairment. In retinal ganglion cells, light responses driven by surviving photoreceptors are obscured by elevated levels of aberrant spiking activity. Here, we demonstrate in rd10 mice that targeting disruptive neuronal circuitry with a gap junction antagonist can significantly reduce excessive spiking. This treatment increases the sensitivity of the degenerated retina to light stimuli driven by residual photoreceptors. Additionally, this enhances signal transmission from inner retinal neurons to ganglion cells, potentially allowing the retinal network to preserve the fidelity of signals either from prosthetic electronic devices, or from cells optogenetically modified to transduce light. Thus, targeting maladaptive changes to the retina allows for treatments to use existing neuronal tissue to restore light sensitivity, and to augment existing strategies to replace lost photoreceptors.

Block of gap junctions eliminates aberrant activity and restores light responses during retinal degeneration.

Spontaneous rhythmic activity is a hallmark feature of the developing retina, where propagating retinal waves instruct axonal targeting and synapse formation. Retinal waves cease around the time of eye-opening; however, the fate of the underlying synaptic circuitry is unknown. Whether retinal waves are unique to the developing retina or if they can be induced in adulthood is not known. Combining patch-clamp techniques with calcium imaging, we demonstrate that propagative events persist in adult mouse retina when it is deprived of inhibitory input. This activity originates in bipolar cells, resembling glutamatergic stage III retinal waves. We find that, as it develops, the network interactions progressively curtail this activity. Together, this provides evidence that the correlated propagative neuronal activity can be induced in adult retina following the blockade of inhibitory interactions.

Botir T Sagdullaev

Ophthalmology

Weill Medical College of Cornell University

Optimized Protocol for Retinal Wholemount Preparation for Imaging and Immunohistochemistry