Stereotactic Injection of MicroRNA-expressing Lentiviruses to the Mouse Hippocampus CA1 Region and Assessment of the Behavioral Outcome

MicroRNAs (miRNAs) are small regulatory single-stranded RNA molecules around 22 nucleotides long that may each target numerous mRNA transcripts and dim an ...

The Hebrew University of Jerusalem

Do contrasting neuropathic pain diagnoses share common pathophysiological mechanisms? Selective breeding was used to derive rat lines with a common genetic background but a striking difference in the degree of spontaneous pain behavior expressed in the neuroma model of neuropathic pain (HA rats (high autotomy) and LA rats (low autotomy)). The contrasting pain phenotype in these lines is attributable to allelic differences at a small number of genetic loci. Here we show that HA and LA rats also differ in their nocifensive response to applied stimuli in the Chung (spinal nerve ligation, SNL) model of neuropathic pain. This includes tactile allodynia and hyperalgesia, and heat allodynia. The degree of hypersensibility varied with sex, age at the time of nerve injury, and the extent of the nerve lesion. F1 crosses of HA and LA rats and inbred Lewis rats showed low levels of autotomy but variable levels of hypersensibility to applied stimuli. Results indicate that alleles which predispose to spontaneous neuropathic pain also predispose to stimulus-evoked pain (allodynia and hyperalgesia). This, in turn, suggests that despite contrasting etiology and behavioral endpoints, pain phenotype in the neuroma and the SNL models shares common pathophysiological mechanisms.

Unity vs. diversity of neuropathic pain mechanisms: Allodynia and hyperalgesia in rats selected for heritable predisposition to spontaneous pain.

Faire contrast&eacute;es de la douleur neuropathique Diagnostics part m&eacute;canismes physiopathologiques communs ? La reproduction s&eacute;lective a &eacute;t&eacute; utilis&eacute;e pour obtenir des lign&eacute;es de rats avec un fond g&eacute;n&eacute;tique commun mais une diff&eacute;rence frappante dans la mesure du comportement de douleur spontan&eacute;e exprim&eacute;e dans le mod&egrave;le de neurinome de la douleur neuropathique (HA rats (autotomie &eacute;lev&eacute;) et LA rat (faible autotomie)). Le ph&eacute;notype de douleur contrast&eacute;es dans ces lignes est attribuable aux diff&eacute;rences all&eacute;liques &agrave; un petit nombre de loci g&eacute;n&eacute;tiques. Ici, nous montrons que les rats HA et LA diff&egrave;rent aussi dans leur r&eacute;ponse de nocifensive aux stimuli appliqu&eacute;s dans le Chung (ligature du nerf spinal, SNL) mod&egrave;le de la douleur neuropathique. Cela inclut l&#39;allodynie tactile et hyperalg&eacute;sie et chaleur allodynie. Le degr&eacute; d&#39;hypersensibilit&eacute; varie avec le sexe, l&#39;&acirc;ge au moment de la l&eacute;sion nerveuse et l&#39;&eacute;tendue de la l&eacute;sion du nerf. F1 traverse des rats HA et LA et des rats Lewis a montr&eacute; de faibles niveaux d&#39;autotomie mais des niveaux variables de l&#39;hypersensibilit&eacute; aux stimuli appliqu&eacute;s consanguins. Les r&eacute;sultats indiquent que les all&egrave;les qui pr&eacute;disposent &agrave; la douleur neuropathique spontan&eacute;e &eacute;galement pr&eacute;disposent &agrave; la douleur induite par stimulation (allodynie et hyperalg&eacute;sie). Cela, &agrave; son tour, laisse entendre que, malgr&eacute; l&#39;&eacute;tiologie contrast&eacute;e et des points de terminaison comportementales, ph&eacute;notype de douleur dans le neurinome et les m&eacute;canismes physiopathologiques communs SNL de mod&egrave;les actions.

Unit&eacute; et diversit&eacute; des m&eacute;canismes de la douleur neuropathique : allodynie et hyperalg&eacute;sie chez les rats s&eacute;lectionn&eacute;s pour une pr&eacute;disposition h&eacute;r&eacute;ditaire &agrave; la douleur spontan&eacute;e.

Fare diagnosi di dolore neuropatico Condividi meccanismi fisiopatologici comuni a contrasto? Allevamento selettivo &egrave; stato utilizzato per derivare linee di ratto con un background genetico comune ma una differenza enorme nel grado di comportamento spontaneo dolore espressa nel modello neuroma di dolore neuropatico (ratti HA (alta Autotomia) e LA ratti (bassa Autotomia)). Il fenotipo di dolore contrastanti in queste righe &egrave; attribuibile alle differenze alleliche a un piccolo numero di loci genetici. Qui vi mostriamo che HA e LA ratti anche differiscono nella loro nocifensive in risposta a stimoli applicati in Chung (legatura nervo spinale, SNL) modello di dolore neuropatico. Questo include esso tattile e iperalgesia e calore esso. Il grado di odontoiatria variata con il sesso, l&#39;et&agrave; al momento della lesione del nervo e la misura della lesione nervosa. F1 incroci di ratti HA e LA e inbred di ratti Lewis hanno mostrati bassi livelli di Autotomia ma livelli variabili di odontoiatria a stimoli applicati. I risultati indicano che gli alleli che predispongono al dolore neuropatico spontaneo anche predispongono a dolore evocato da stimolo (interessato e iperalgesia). Questo, a sua volta, suggerisce che nonostante eziologia contrasto e comportamentale endpoint, fenotipo dolore il neuroma e il SNL modelli azioni comuni meccanismi fisiopatologici.

Unit&agrave; vs diversit&agrave; dei meccanismi del dolore neuropatico: esso e iperalgesia nei ratti selezionati per predisposizione ereditaria al dolore spontaneo.

Exercise and inherited factors both affect recovery from stroke and head injury, but the underlying mechanisms and interconnections between them are yet unknown. Here, we report that similar cation channels mediate the protective effect of exercise and specific genetic background in a kainate injection model of cerebellar stroke. Microinjection to the cerebellum of the glutamatergic agonist, kainate, creates glutamatergic excito\xE2\x80\x90toxicity characteristic of focal stroke, head injury or alcoholism. Inherited protection and prior exercise were both accompanied by higher cerebellar expression levels of the Kir6.1 ATP-dependent potassium channel in adjacent Bergmann glia, and voltage-gated KVbeta2 and cyclic nucleotide-gated cation HCN1 channels in basket cells. Sedentary FVB/N and exercised C57BL/6 mice both expressed higher levels of these cation channels compared to sedentary C57BL/6 mice, and were both found to be less sensitive to glutamate toxicity. Moreover, blocking ATP-dependent potassium channels with Glibenclamide enhanced kainate-induced cell death in cerebellar slices from the resilient sedentary FVB/N mice. Furthermore, exercise increased the number of acetylcholinesterase-positive fibres in the molecular layer, reduced cerebellar cytokine levels and suppressed serum acetylcholinesterase activity, suggesting anti-inflammatory protection by enhanced cholinergic signalling. Our findings demonstrate for the first time that routine exercise and specific genetic backgrounds confer protection from cerebellar glutamatergic damages by similar molecular mechanisms, including elevated expression of cation channels. In addition, our findings highlight the involvement of the cholinergic anti-inflammatory pathway in insult-inducible cerebellar processes. These mechanisms are likely to play similar roles in other brain regions and injuries as well, opening new venues for targeted research efforts.

Similar cation channels mediate protection from cerebellar exitotoxicity by exercise and inheritance.

Exercice et facteurs h&eacute;r&eacute;ditaires les deux affectent la r&eacute;cup&eacute;ration de blessures accident vasculaire c&eacute;r&eacute;bral et de la t&ecirc;te, mais les m&eacute;canismes sous-jacents et les interconnexions entre eux sont encore inconnus. Nous rapportons ici que des canaux cationiques similaires m&eacute;dient l&#39;effet protecteur de l&#39;exercice et les fond g&eacute;n&eacute;tique sp&eacute;cifique dans un mod&egrave;le d&#39;injection de ka&iuml;nate d&#39;AVC c&eacute;r&eacute;belleux. Microinjection au cervelet de l&#39;agoniste glutamatergiques ka&iuml;nate, cr&eacute;e glutamatergique excito\xE2\x80\x90toxicity caract&eacute;ristique de focale accident vasculaire c&eacute;r&eacute;bral, traumatisme cr&acirc;nien ou d&#39;alcoolisme. Protection h&eacute;rit&eacute;e et exercice pr&eacute;alable &eacute;taient tous deux accompagn&eacute;s d&#39;expression c&eacute;r&eacute;belleux sup&eacute;rieurs le canal potassique Kir6.1 ATP-d&eacute;pendante dans adjacente Bergmann glia et KVbeta2 voltage-d&eacute;pendants et les canaux de cation de d&eacute;coupage en nucl&eacute;otides cycliques HCN1 dans les cellules de panier. S&eacute;dentaire FVB/N et des souris C57BL/6 lev&eacute;es les deux exprim&eacute; des niveaux plus &eacute;lev&eacute;s de ces canaux cationiques par rapport aux s&eacute;dentaires souris C57BL/6, et tous deux se sont av&eacute;r&eacute;s pour &ecirc;tre moins sensibles &agrave; la toxicit&eacute; du glutamate. En outre, blocage des canaux potassiques ATP-d&eacute;pendants avec le Glibenclamide am&eacute;lior&eacute; la mort cellulaire induite par le ka&iuml;nate dans tranches de cervelet des souris FVB/N s&eacute;dentaires r&eacute;silients. En outre, exercice a augment&eacute; le nombre de fibres de l&#39;ac&eacute;tylcholinest&eacute;rase positifs dans la couche mol&eacute;culaire, r&eacute;duit les niveaux de cytokine c&eacute;r&eacute;belleux et supprim&eacute; l&#39;activit&eacute; de l&#39;ac&eacute;tylcholinest&eacute;rase de s&eacute;rum, sugg&eacute;rant des anti-inflammatoires protection de signalisation cholinergiques am&eacute;lior&eacute;e. Nos r&eacute;sultats d&eacute;montrent pour la premi&egrave;re fois que l&#39;exercice routinier et fonds g&eacute;n&eacute;tiques sp&eacute;cifiques conf&egrave;rent protection contre les dommages c&eacute;r&eacute;belleux glutamatergique par des m&eacute;canismes mol&eacute;culaires semblables, y compris expression &eacute;lev&eacute;e des canaux cationiques. En outre, nos r&eacute;sultats mettent en &eacute;vidence l&#39;implication de la voie cholinergique anti-inflammatoires dans les processus c&eacute;r&eacute;belleuses insultes inductibles. Ces m&eacute;canismes sont susceptibles de jouer un r&ocirc;le similaire dans d&#39;autres r&eacute;gions du cerveau et des blessures ainsi, ouverture de nouveaux sites pour les efforts de recherche cibl&eacute;s.

Des canaux cationiques similaires m&eacute;dient protection contre exitotoxicity c&eacute;r&eacute;belleux par exercice et h&eacute;ritage.

Esercizio sia fattori ereditari possono influenzare recupero da ictus e testa ferita, ma i meccanismi e le interconnessioni tra di loro sottostanti sono ancora sconosciuto. Qui, segnaliamo che simili canali cazione mediano l&#39;effetto protettivo di esercizio e patrimonio genetico specifico in un modello ad iniezione Kainato stroke cerebellare. Microinjection al cervelletto di agonista glutamatergico, Kainato, crea glutamatergico excito\xE2\x80\x90toxicity caratteristica focale ictus, trauma cranico o alcolismo. Protezione ereditati ed esercizio precedente erano accompagnate da livelli pi&ugrave; elevati di espressione cerebellare di cyclic nucleotide-gated cazione HCN1 canali nelle cellule del cestino e il canale del potassio Kir6.1 trifosfato di Adenosina-dipendente nelle adiacente glia di Bergmann e KVbeta2 di tensione-gated. FVB/N sedentario e topi C57BL/6 esercitati sia espressero livelli pi&ugrave; elevati di questi canali cazione rispetto ai topi C57BL/6 sedentari, ed entrambi sono stati trovati per essere meno sensibili alla tossicit&agrave; del glutammato. Inoltre, bloccando i canali potassio trifosfato di Adenosina-dipendente con Glibenclamide enhanced morte cellulare indotta da Kainato cerebellare fette dai topi sedentari resilienti FVB/N. Inoltre, esercizio aumentato il numero di fibre dell&#39;acetilcolinesterasi positivo nello strato molecolare, ha ridotto i livelli di citochine cerebellare e soppresso l&#39;attivit&agrave; dell&#39;acetilcolinesterasi sierica, suggerendo protezione anti-infiammatori di segnalazione avanzata colinergici. I nostri risultati dimostrano per la prima volta che routine di esercizio e background genetico specifico conferisce protezione da danni glutamatergico cerebellare da simili meccanismi molecolari, tra cui elevata espressione dei canali del catione. Inoltre, i nostri risultati evidenziano il coinvolgimento del percorso antinfiammatorio colinergico in insulto-inducible processi cerebellari. Questi meccanismi sono propensi a giocare ruoli simili in altre regioni cerebrali e lesioni pure, aprendo nuove sedi per gli sforzi di ricerca mirata.

Simili canali cazione mediano protezione da exitotoxicity cerebellare di esercizio e l&#39;eredit&agrave;.

Cholinergic signaling suppresses inflammation in blood and brain and attenuates apoptosis in other tissues, but whether it blocks inflammation in skeletal muscle under toxicant exposure, injuries and diseases remained unexplored. Here, we report nicotinic attenuation of inflammation and alteration of apoptotic protein expression pattern in murine muscle tissue and cultured myotubes, involving the RNA-binding protein, Tristetraprolin, and the anti-apoptotic protein, Mcl-1. In muscles and C2C12 myotubes, cholinergic excitation by exposure to nicotine or the organophosphorous pesticide, Paraoxon, induced Tristetraprolin overproduction while reducing pro-inflammatory transcripts such as IL-6, CXCL1 (KC) and CCL2 (MCP-1). Furthermore, nicotinic excitation under exposure to the bacterial endotoxin LPS attenuated over-expression of the CCL2 and suppressed the transcriptional activity of NF-Ä¸B and AP-1. Tristetraprolin was essential for this anti-inflammatory effect of nicotine in basal conditions. However, its knockdown also impaired the pro-inflammatory response to LPS. Finally, in vivo administration of Paraoxon or recombinant Acetylcholinesterase, leading respectively to either gain or loss of cholinergic signaling, modified muscle expression of key mRNA processing factors and several of their apoptosis-related targets. Specifically, cholinergic imbalances enhanced the kinase activators of the Serine-Arginine splicing kinases, Clk1 and Clk3. Moreover, Paraoxon raised the levels of the anti-apoptotic protein, Mcl-1, through a previously unrecognized polyadenylation site selection mechanism, producing longer, less stable Mcl-1 mRNA transcripts. Together, our findings demonstrate that in addition to activating muscle function, acetylcholine regulates muscle inflammation and cell survival, and point to Tristetraprolin and the choice of Mcl-1 mRNA polyadenylation sites as potential key players in muscle reactions to insults.

Nicotinic stimulation induces Tristetraprolin over-production and attenuates inflammation in muscle.

Signalisation cholinergiques supprime l&#39;inflammation dans le sang et le cerveau et att&eacute;nue l&#39;apoptose dans d&#39;autres tissus, mais si il bloque l&#39;inflammation dans le muscle squelettique au titre de l&#39;exposition toxique, blessures et maladies sont rest&eacute;s inexplor&eacute;s. Nous rapportons ici, nicotinique att&eacute;nuation de l&#39;inflammation et la modification du mod&egrave;le d&#39;expression de prot&eacute;ine apoptotique dans le tissu musculaire murine et cultiv&eacute;es myotubes, impliquant la prot&eacute;ine de liaison &agrave; l&#39;ARN, Tristetraprolin et la prot&eacute;ine anti-apoptotique, Mcl-1. Dans les muscles et les myotubes C2C12, excitation cholinergique par exposition &agrave; la nicotine ou les pesticides organophosphor&eacute;s, Paraoxon, induite par le surproduction de tristetraprolin tout en r&eacute;duisant les transcriptions pro-inflammatoires telles que l&#39;IL-6, CXCL1 (KC) et CCL2 (MCP-1). En outre, excitation nicotinique sous l&#39;exposition &agrave; l&#39;endotoxine bact&eacute;rienne LPS a att&eacute;nu&eacute; la surexpression de la CCL2 et supprim&eacute; l&#39;activit&eacute; transcriptionnelle de NF-&#x138;B et AP-1. Tristetraprolin a &eacute;t&eacute; essentiel pour cet effet anti-inflammatoire de la nicotine dans des conditions basales. Cependant, sa pr&eacute;cipitation r&eacute;duite &eacute;galement la r&eacute;ponse pro-inflammatoire LPS. Enfin, l&#39;administration in vivo de Paraoxon ou recombinante ac&eacute;tylcholinest&eacute;rase, conduisant respectivement &agrave; gain ou perte de signal cholinergique, modifi&eacute;e expression musculaire des facteurs cl&eacute;s mRNA processing et plusieurs de leurs cibles li&eacute;es &agrave; l&#39;apoptose. Plus pr&eacute;cis&eacute;ment, les d&eacute;s&eacute;quilibres cholinergiques am&eacute;lior&eacute; les activateurs de la kinase des kinases &eacute;pissage s&eacute;rine-Arginine, Clk1 et Clk3. En outre, Paraoxon &eacute;lev&eacute; les niveaux de la prot&eacute;ine anti-apoptotique, Mcl-1, gr&acirc;ce &agrave; un m&eacute;canisme de s&eacute;lection de site de polyad&eacute;nylation pr&eacute;c&eacute;demment non reconnu, produire plus, moins stable ARNm de Mcl-1. Ensemble, nos r&eacute;sultats d&eacute;montrent qu&#39;en plus de l&#39;activation de la fonction musculaire, l&#39;ac&eacute;tylcholine r&eacute;glemente survie de l&#39;inflammation et de la cellule de muscle et point &agrave; Tristetraprolin et le choix des sites de polyad&eacute;nylation ARNm de Mcl-1 comme d&#39;&eacute;ventuels participants cl&eacute;s dans les r&eacute;actions musculaires aux insultes.

Stimulation nicotinique induit une surproduction de Tristetraprolin et att&eacute;nue l&#39;inflammation dans les muscles.

Segnalazione colinergici sopprime l&#39;infiammazione nel sangue e cervello e attenua il apoptosis in altri tessuti, ma se si blocca l&#39;infiammazione nel muscolo scheletrico sotto esposizione misurino, lesioni e malattie &egrave; rimasto inesplorate. Qui, segnaliamo nicotinici attenuazione dell&#39;infiammazione e l&#39;alterazione del modello di espressione della proteina apoptotic in tessuto muscolare murino e myotubes colta, che coinvolgono il RNA-binding protein, Tristetraprolin e la proteina anti-apoptotic, Mcl-1. Nei muscoli e C2C12 myotubes, eccitazione colinergica da esposizione alla nicotina o gli antiparassitari organofosforici, Paraoxon, indotta da sovrapproduzione Tristetraprolin riducendo le trascrizioni pro-infiammatorie come IL-6, CXCL1 (KC) e CCL2 (MCP-1). Inoltre, nicotinici eccitazione in esposizione per la LPS di endotossine batteriche attenuato sovraespressione della CCL2 e soppresso l&#39;attivit&agrave; trascrizionale di NF-&#x138;B e AP-1. Tristetraprolin &egrave; stato essenziale per questo effetto antinfiammatorio di nicotina in condizioni basali. Tuttavia, l&#39;atterramento anche alterata la risposta pro-infiammatoria a LPS. Infine, somministrazione in vivo di Paraoxon o ricombinante dell&#39;acetilcolinesterasi, che portano rispettivamente a guadagno o perdita di segnalazione colinergici, modificato muscolo espressione di fattori chiave elaborazione mRNA e molti dei loro obiettivi correlati a apoptosis. In particolare, gli squilibri colinergici enhanced gli attivatori della chinasi della chinasi di splicing serina-arginina, Clk1 e Clk3. Inoltre, Paraoxon ha generato i livelli delle proteine anti-apoptotic, Mcl-1, attraverso un meccanismo di selezione del sito in precedenza non riconosciuta poliadenilazione, producendo pi&ugrave;, meno stabile trascritti di mRNA Mcl-1. Insieme, i nostri risultati dimostrano che oltre ad attivare la funzione muscolare, acetilcolina regola sopravvivenza infiammazione e delle cellule muscolari e punto di Tristetraprolin e la scelta dei siti di poliadenilazione Mcl-1 mRNA come potenziali giocatori chiave nelle reazioni muscolari agli insulti.

Nicotina stimolazione induce Tristetraprolin sovrapproduzione e attenua l&#39;infiammazione nel muscolo.

End-stage Alzheimer's disease (AD) involves drastic modifications in neuronal molecular and cellular processes, but little is known about the dynamics of these modifications during disease initiation and progression. Here, we report meta-analysis of 100 publicly available Microarray datasets using threshold-independent analysis. We found that different patients react to AD progression by variable single transcript alterations which however lead to similar changes in functional gene groups. Stratification by patients' cognitive deterioration pre`sented hippocampal-specific mRNA alterations which involved progressively changed gene categories and indicate changes in epigenetic state and microRNA profiles. In addition, datasets from laser-captured neurofibrillary tangles-free hippocampal neurons and transcript classification by cell types identified many of these changes in neurons. Intriguingly, we discovered that early-onset decline in alternative splicing, protein folding and transport transcripts occur concurrently with decreases in synaptic transmission, whereas at later stages these changes progressed into enhanced oxidative stress and inflammation. Our findings open new venues for identifying novel targets for intervention with AD progression.

Threshold-independent meta-analysis of Alzheimer's disease transcriptomes shows progressive changes in hippocampal functions, epigenetics and microRNA regulation.

Terminale maladie d&#39;Alzheimer (ma) implique des modifications drastiques dans les processus mol&eacute;culaires et cellulaires neuronaux, mais on en sait peu sur la dynamique de ces modifications au cours de la progression et l&#39;initiation de la maladie. Nous rapportons ici, meta-analysis of 100 de donn&eacute;es Microarray publiquement disponibles en utilisant l&#39;analyse ind&eacute;pendante du seuil. Nous avons constat&eacute; que diff&eacute;rents patients r&eacute;agissent &agrave; la progression de l&#39;AD par des modifications de transcription seule variable qui toutefois conduisent &agrave; des changements similaires dans les groupes de g&egrave;ne fonctionnel. Stratification des patients d&eacute;t&eacute;rioration cognitive pre&#39;sented hippocampe sp&eacute;cifiques ARNm alt&eacute;rations entra&icirc;nant progressivement chang&eacute; cat&eacute;gories de g&egrave;ne et indiquer les changements &eacute;pig&eacute;n&eacute;tiques profils d&#39;&Eacute;tat et des micro-ARN. En outre, ensembles de laser-captur&eacute; enchev&ecirc;trements neurofibrillaires libres neurones de l&#39;hippocampe et classification de la transcription par types de cellules identifi&eacute; plusieurs de ces changements dans les neurones. Curieusement, nous avons d&eacute;couvert que le d&eacute;clin pr&eacute;coce dans l&#39;&eacute;pissage alternatif, transcriptions pliage et transport de prot&eacute;ines se produisent en m&ecirc;me temps que les diminutions de la transmission synaptique, alors qu&#39;&agrave; un stade ult&eacute;rieur, ces changements ont progress&eacute; vers l&#39;inflammation et du stress oxydatif am&eacute;lior&eacute;. Nos r&eacute;sultats ouvrent de nouveaux lieux pour l&#39;identification de nouvelles cibles d&#39;intervention avec la progression de l&#39;AD.

Ind&eacute;pendant du seuil m&eacute;ta-analyse de la maladie d&#39;Alzheimer transcriptomes montre des changements progressifs dans les fonctions de l&#39;hippocampe, r&eacute;gulation &eacute;pig&eacute;n&eacute;tique et micro-ARN.

Stadio terminale della malattia di Alzheimer (AD) comporta modifiche drastiche in processi molecolari e cellulari neuronali, ma poco si sa circa le dinamiche di queste modifiche durante la malattia iniziazione e progressione. Qui, segnaliamo la meta-analisi di 100 disponibili pubblicamente Microarray DataSet mediante analisi indipendente dalla soglia. Abbiamo scoperto che diversi pazienti reagiscono alla progressione AD alterazioni di variabile singola trascrizione che tuttavia portano a cambiamenti simili nei gruppi del gene funzionale. Stratificazione di deterioramento cognitivo pre&#39;sented hippocampal specifici mRNA alterazioni pazienti che ha coinvolto progressivamente cambiato categorie gene e indicare cambiamenti epigenetici profili dello stato e microRNA. Inoltre, DataSet da trascrizione classificazione di tipi di cellule e neuroni hippocampal senza grovigli neurofibrillary laser-catturato identificati molti di questi cambiamenti nei neuroni. Curiosamente, abbiamo scoperto che il declino precoce di splicing alternativo, le trascrizioni di piegatura e trasporto della proteina si verificano contemporaneamente diminuisce nella trasmissione sinaptica, considerando che nelle fasi successive queste modifiche progredito nell&#39;infiammazione e stress ossidativo rafforzato. I nostri risultati aprire nuove sedi per l&#39;identificazione di nuovi bersagli per l&#39;intervento con la progressione della AD.

Indipendente dalla soglia di meta-analisi della malattia di Alzheimer transcriptomes illustrato progressivi cambiamenti nelle funzioni dell&#39;ippocampo, regolazione epigenetica e microRNA.

Genetic studies link inherited errors in RNA metabolism to familial neurodegenerative disease. Here, we report such errors and the underlying mechanism in sporadic Alzheimer's disease (AD). AD entorhinal cortices presented globally impaired exon exclusions and selective loss of the hnRNP A/B splicing factors. Supporting functional relevance, hnRNP A/B knockdown induced alternative splicing impairments and dendrite loss in primary neurons, and memory and electrocorticographic impairments in mice. Transgenic mice with disease-associated mutations in APP or Tau displayed no alterations in hnRNP A/B suggesting that its loss in AD is independent of AÎ² and Tau toxicity. However, cholinergic excitation increased hnRNP A/B levels while in vivo neurotoxin-mediated destruction of cholinergic neurons caused cortical AD-like decrease in hnRNP A/B and recapitulated the alternative splicing pattern of AD patients. Our findings present cholinergic-mediated hnRNP A/B loss and impaired RNA metabolism as important mechanisms involved in AD.

Cholinergic-associated loss of hnRNP-A/B in Alzheimer's disease impairs cortical splicing and cognitive function in mice.

Des &eacute;tudes g&eacute;n&eacute;tiques lien Erreurs h&eacute;rit&eacute;es dans le m&eacute;tabolisme de RNA de maladie neurod&eacute;g&eacute;n&eacute;rative familial. Nous rapportons ici, ces erreurs et le m&eacute;canisme sous-jacent dans la maladie d&#39;Alzheimer sporadique (AD). Cortex entorhinal AD pr&eacute;sent&eacute; globalement alt&eacute;r&eacute;e exon exclusions et perte s&eacute;lective de la hnRNP A / B facteurs d&#39;&eacute;pissage. Soutenir la pertinence fonctionnelle, hnRNP A / knockdown B induite par des d&eacute;ficiences &eacute;pissage alternatifs et la perte de la dendrite dans des neurones primaires et d&eacute;ficiences de m&eacute;moire et electrocorticographic chez la souris. Des souris transg&eacute;niques pr&eacute;sentant des mutations associ&eacute;es &agrave; la maladie dans l&#39;APP ou Tau n&#39;affichent aucune alt&eacute;ration dans hnRNP A / B, sugg&eacute;rant que sa perte en AD est ind&eacute;pendant de toxicit&eacute; A&beta; et Tau. Cependant, excitation cholinergique a augment&eacute; A de hnRNP / B niveaux tandis qu&#39;in vivo neurotoxine induite par la destruction des neurones cholinergiques a provoqu&eacute; corticale AD-comme diminution hnRNP A / B et r&eacute;capitul&eacute; le patron &eacute;pissage alternatif des patients atteints de ma. Nos r&eacute;sultats pr&eacute;sentent hnRNP m&eacute;diation cholinergique A / B perte et m&eacute;tabolisme de RNA douteux comme des m&eacute;canismes importants impliqu&eacute;s dans AD.

Cholinergiques associ&eacute;es &agrave; la perte des hnRNP-A/B dans la maladie d&#39;Alzheimer alt&egrave;re l&#39;&eacute;pissage corticale et la fonction cognitive chez les souris.

Studi genetici link errori ereditati nel metabolismo del RNA malattia neurodegenerativa familiare. Riportiamo tali errori ed il meccanismo sottostante in sporadico della malattia di Alzheimer (AD). Cortecce entorinale annuncio presentato globalmente compromessa esone esclusioni e perdita selettiva di hnRNP A / B fattori di splicing. Supporto di pertinenza funzionale, hnRNP A / atterramento B indotta da menomazioni splicing alternativi e perdita di dendrite in neuroni primari e disturbi della memoria ed electrocorticographic nei topi. Topi transgenici con mutazioni associate a malattie APP o Tau non visualizzato alterazioni in hnRNP A / B, suggerendo che la sua perdita in annuncio &egrave; indipendente di tossicit&agrave; A&beta; e Tau. Tuttavia, eccitazione colinergico aumentato hnRNP A / B livelli mentre in vivo neurotossina-mediata distruzione dei neuroni colinergici causata corticale diminuzione simile AD hnRNP A / B e ricapitolato il pattern di splicing alternativo dei pazienti AD. I nostri risultati presentano hnRNP colinergica mediata A / B perdita e alterato metabolismo del RNA come meccanismi importanti coinvolti nell&#39;annuncio.

Colinergica associata perdita di hnRNP-A/B nella malattia di Alzheimer altera lo splicing corticale e le funzioni cognitive nei topi.

Classification analysis based on high throughput data is a common feature in neuroscience and other fields of science, with a rapidly increasing impact on both basic biology and disease-related studies. The outcome of such classifications often serves to delineate novel biochemical mechanisms in health and disease states, identify new targets for therapeutic interference, and develop innovative diagnostic approaches. Given the importance of this type of studies, we screened 111 recently-published high-impact manuscripts involving classification analysis of gene expression, and found that 58 of them (53%) based their conclusions on a statistically invalid method which can lead to bias in a statistical sense (lower true classification accuracy then the reported classification accuracy). In this report we characterize the potential methodological error and its scope, investigate how it is influenced by different experimental parameters, and describe statistically valid methods for avoiding such classification mistakes.

Statistically invalid classification of high throughput gene expression data.

Analyse de classification bas&eacute;e sur les donn&eacute;es &agrave; haut d&eacute;bit est une caract&eacute;ristique courante dans les neurosciences et d&#39;autres domaines de la science, avec un impact augmente rapidement sur biologie fondamentale et &eacute;tudes li&eacute;s &agrave; la maladie. Le r&eacute;sultat de ces classifications sert souvent &agrave; d&eacute;limiter des nouveaux m&eacute;canismes biochimiques dans les &Eacute;tats de sant&eacute; et la maladie, identifier de nouvelles cibles d&#39;intervention th&eacute;rapeutique et d&eacute;velopper des approches innovantes de Diagnostics. Compte tenu de l&#39;importance de ce type d&#39;&eacute;tudes, les auteurs ont cribl&eacute; 111 publi&eacute; r&eacute;cemment percutantes manuscrits impliquant l&#39;analyse de la classification de l&#39;expression g&eacute;nique et trouv&eacute; que 58 d&#39;entre eux (53 %) fond&eacute; leurs conclusions sur une m&eacute;thode non statistiquement valide qui peut conduire &agrave; des biais dans un sens statistique (exactitude de v&eacute;ritable classification inf&eacute;rieure puis la pr&eacute;cision de classification signal&eacute;s). Dans le pr&eacute;sent rapport que nous caract&eacute;riser l&#39;erreur m&eacute;thodologique possible et sa port&eacute;e, &eacute;tudier comment elle est influenc&eacute;e par diff&eacute;rents param&egrave;tres exp&eacute;rimentaux et d&eacute;crire des m&eacute;thodes statistiquement valables pour &eacute;viter de telles erreurs de classification.

Classification non statistiquement valide de donn&eacute;es d&#39;expression de g&egrave;nes &agrave; haut d&eacute;bit.

Analisi di classificazione sulla base dei dati ad alta velocit&agrave; sono una caratteristica comune nelle neuroscienze e in altri campi della scienza, con un impatto crescente rapidamente sia biologia e studi riguardanti le malattie. L&#39;esito di tali classificazioni spesso serve a delineare nuovi meccanismi biochimici negli Stati di salute e malattia, identificare nuovi bersagli terapeutici interferenze e sviluppare approcci diagnostici innovativi. Data l&#39;importanza di questo tipo di studi, abbiamo proiettato 111 recentemente pubblicato ad alto impatto manoscritti che coinvolgono analisi di classificazione dell&#39;espressione genica e trovato che 58 di loro (53%) basato le loro conclusioni su un metodo statisticamente non valido che pu&ograve; portare alla polarizzazione in senso statistico (minore accuratezza della classificazione vero poi l&#39;accuratezza di classificazione riferito). In questa relazione che ci caratterizzano i potenziali errori metodologici e la sua portata, indagare come &egrave; influenzata da diversi parametri sperimentali e descrivere statisticamente validi metodi per evitare tali errori di classificazione.

Shahar Barbash

Department of Biological Chemistry and The Edmond & Lily Safra Center for Brain Sciences

The Hebrew University of Jerusalem

Stereotactic Injection of MicroRNA-expressing Lentiviruses to the Mouse Hippocampus CA1 Region and Assessment of the Behavioral Outcome

Shahar Barbash

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Biological Chemistry and The Edmond & Lily Safra Center for Brain Sciences

The Hebrew University of Jerusalem

Stereotactic Injection of MicroRNA-expressing Lentiviruses to the Mouse Hippocampus CA1 Region and Assessment of the Behavioral Outcome

Department of Biological Chemistry and The Edmond & Lily Safra Center for Brain Sciences