Name: identification of dopamine d1-alpha receptor within rodent nucleus accumbens by an innovative rna in situ detection technology
Uploaded: 2018-03-27T15:00:00
Description: Watch this Scientific Journal Video about Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology at JoVE.com

Le opere presenti sono state sostenute dagli istituti nazionali di sovvenzioni di salute (NIH) HD043680, MH106392, DA013137 e NS100624. Fondo parziale è stato fornito da una sovvenzione di formazione NIH T32 nella scienza biomedica-comportamentale.

Il protocollo sperimentale è stato approvato dalla cura degli animali e uso Committee (IACUC) presso la University of South Carolina (numero di assicurazione federale: A3049-01).
1. preparazione di sezioni di cervello congelato fresco
<ol><li>Utilizzare il ceppo di ratti F344/N: tre ratti di ogni sesso, Guida in linea di 13 mesi di età, circa 320 g di peso corporeo.</li>
	<li>Regolare la concentrazione di sevoflurane al 5% (sovradosaggio del sevoflurano). Continuare l'esposizione sevoflura...

<ol>
	<li>Goldman-Rakic, P. S., Castner, S. A., Svensson, T. H., Siever, L. J., Williams, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+the+dopamine+D1+receptor+in+schizophrenia:+insights+for+cognitive+dysfunction.">Targeting the dopamine D1 receptor in schizophrenia: insights for cognitive dysfunction.</a> Psychopharmacology (Berl). 174 (1), 3-16 (2004).</li><li>Beaulieu, J. M., Gainetdinov, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+physiology,+signaling,+and+pharmacology+of+dopamine+receptors.">The physiology, signaling, and pharmacology of dopamine receptors.</a> Pharmacol Rev. 63, 182-217 (2011).</li><li>Zahrt, J., Taylor, J. R., Mathew, R. G., Arnsten, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Supranormal+stimulation+of+D1+dopamine+receptors+in+the+rodent+prefrontal+cortex+impairs+spatial+working+memory+performance.">Supranormal stimulation of D1 dopamine receptors in the rodent prefrontal cortex impairs spatial working memory performance.</a> J Neurosci. 17, 8528-8535 (1997).</li><li>Floresco, S. B., Magyar, O., Ghods-Sharifi, S., Vexelman, C., Tse, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+dopamine+receptor+subtypes+in+the+medial+prefrontal+cortex+of+the+rat+regulate+set-shifting.">Multiple dopamine receptor subtypes in the medial prefrontal cortex of the rat regulate set-shifting.</a> Neuropsychopharmacology. 31, 297-309 (2006).</li><li>Arnsten, A. F., Girgis, R. R., Gray, D. L., Mailman, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+dopamine+therapeutics+for+cognitive+deficits+in+schizophrenia.">Novel dopamine therapeutics for cognitive deficits in schizophrenia.</a> Biol Psychiatry. 81, 67-77 (2017).</li><li>Ellenbroek, B. A., Budde, S., Cools, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prepulse+inhibition+and+latent+inhibition:+the+role+of+dopamine+in+the+medial+prefrontal+cortex.">Prepulse inhibition and latent inhibition: the role of dopamine in the medial prefrontal cortex.</a> Neuroscience. 75 (2), 535-542 (1996).</li><li>Parker, K. L., Alberico, S. L., Miller, A. D., Narayanan, N. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prefrontal+D1+dopamine+signaling+is+necessary+for+temporal+expectation+during+reaction+time+performance.">Prefrontal D1 dopamine signaling is necessary for temporal expectation during reaction time performance.</a> Neuroscience. 255, 246-254 (2013).</li><li>Manduca, A., Servadio, M., Damsteegt, R., Campolongo, P., Vanderschuren, L. J., Trezza, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dopaminergic+Neurotransmission+in+the+Nucleus+Accumbens+Modulates+Social+Play+Behavior+in+Rats.">Dopaminergic Neurotransmission in the Nucleus Accumbens Modulates Social Play Behavior in Rats.</a> Neuropsychopharmacology. 41 (9), 2215-2223 (2016).</li><li>Okubo, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+prefrontal+dopamine+D1+receptors+in+schizophrenia+revealed+by+PET.">Decreased prefrontal dopamine D1 receptors in schizophrenia revealed by PET.</a> Nature. 385 (6617), 634-636 (1997).</li><li>Abi-Dargham, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prefrontal+dopamine+D1+receptors+and+working+memory+in+schizophrenia.">Prefrontal dopamine D1 receptors and working memory in schizophrenia.</a> J Neurosci. 22 (9), 3708-3719 (2002).</li><li>Shinohara, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dopamine+D1+receptor+subtype+mediates+acute+stress-induced+dendritic+growth+in+excitatory+neurons+of+the+medial+prefrontal+cortex+and+contributes+to+suppression+of+stress+susceptibility+in+mice.">Dopamine D1 receptor subtype mediates acute stress-induced dendritic growth in excitatory neurons of the medial prefrontal cortex and contributes to suppression of stress susceptibility in mice.</a> Mol Psychiatry. 19, (2017).</li><li>Okubo, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+prefrontal+dopamine+D1+receptors+in+schizophrenia+revealed+by+PET.">Decreased prefrontal dopamine D1 receptors in schizophrenia revealed by PET.</a> Nature. 385, 634-636 (1997).</li><li>Wang, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RNAscope:+a+novel+in+situ+RNA+analysis+platform+for+formalin-fixed,+paraffin-embedded+tissues.">RNAscope: a novel in situ RNA analysis platform for formalin-fixed, paraffin-embedded tissues.</a> J Mol Diagn. 14 (1), 22-29 (2012).</li><li>Paxinos, G., Watson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The rat brain in stereotaxic coordinates. , (2014).</li></ol>

In questo protocollo, descriviamo una tecnica novella di ibridazione in situ per fette di cervello fresco congelato per valutare l'espressione del ricevitore Drd1α nel nucleus accumbens (NAc) e l'espressione di tirosina idrossilasi (TH) in regione della substantia nigra (SNR). Forniamo anche i metodi di analisi dei dati per diversi pattern di colorazione: "discreto punto" o "cluster".
I passaggi critici per un'analisi di fluorescenza multisala successo RNA in situ di ibridaz...

Disfunzione del sistema della dopamina striatale è coinvolto nella progressione dei sintomi clinici osservati nelle malattie neurocognitive multiple. I ricevitori della dopamina D1 sono presenti nella corteccia prefrontale (PFC) e regioni striatal del cervello e influenzano pesantemente i processi cognitivi1, compreso il funzionamento memoria, elaborazione temporale e comportamento locomotiva2,3 , 4 , 5 , 6 , 7. precedenti studi chiarito che le modifiche dei ricevitori della dopamina D1 sono state associate con la progressione di attenzione iperattività di deficit disorder (ADHD)8, i sintomi neurocognitivi nella schizofrenia9, 10 e lo stress suscettibilità11. In particolare, nella schizofrenia, tomografia a emissione di positroni (PET) studi hanno indicato che la capacità di legame dei recettori D1 della dopamina nelle cortecce prefrontali era altamente relative al deficit cognitivi e la presenza di sintomi negativi11. La crescita dendritica dei neuroni eccitatori nella corteccia prefrontale regolato dal recettore della dopamina D1 allevia lo stress suscettibilità. Inoltre, l'atterramento del recettore D1 in neuroni corticali prefrontale mediale (mPFC) potrebbe migliorare la sconfitta sociale indotta da stress sociale evitare12.
Qui, presentiamo una nuova tecnica di RNA in situ ibridazione per visualizzare singole molecole di RNA in una cella con campioni di tessuto fresco congelato. La tecnica attuale ha molteplici vantaggi rispetto ai metodi che esistono all'interno della letteratura corrente. In primo luogo, l'attuale procedura per mantenere il contesto spaziale e morfologico del tessuto ed è stata effettuata su campioni di tessuto fresco congelato affinché altre procedure che richiedono fresco, non incorporati tessuti possono essere combinati con i metodi attuali. Procedure analoghe nei tessuti formalina-fisso e paraffina-incastonati hanno illustrato che risoluzione singola trascrizione può essere realizzato usando un RNA in situ ibridazione tecnica13. Rilevamento del RNA a livello di singola trascrizione fornisce una sensibilità superiore alla espressione di numero di copia basso così come l'opportunità di confrontare l'espressione genica a livello di singole cellule che non possono essere raggiunti da altri metodi di rilevazione dell'acido nucleico, quali tecniche di reazione a catena (PCR) della polimerasi. Inoltre, il metodo corrente mantiene immagini con un elevato rapporto segnale-rumore attraverso le sonde del RNA altamente specifiche che sono ibridate di trascritti di RNA bersaglio singolo e associate in sequenza con una cascata di molecole di amplificazione del segnale nella rilevazione sistema. Infine, l'attuale tecnologia offre l'opportunità di valutare più sistemi biologici con sue sonde proprietarie di target-specifici, piuttosto che limitare la nostra ricerca ad una sola classe di indicatori relativi al sistema quali il rilevamento di proteine di metodi di esame immuno-istochimico.
Nel nostro studio, abbiamo usato questa romanzo ibridazione in situ per RNA per valutare l'espressione del ricevitore Drd1α nel nucleus accumbens (NAc) e l'espressione di tirosina idrossilasi (TH) nella substantia nigra (SNR) dei ratti F344/N sia maschili che femminili. L'innovativo ibridazione in situ per RNA permesso di studiare i meccanismi che influenzano sia DA assorbimento e rilascio di DA simultaneamente, migliorando la nostra comprensione della complessità del sistema striatal del DA. Qui, descriviamo la procedura per fette di cervello fresco-congelato e fornire i metodi di analisi dei dati per diversi pattern di colorazione: "discreto punto" o "cluster".

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	<tbody>
		<tr height="23">
			<td height="23" style="height:23px;width:329px;">HybEZ Oven system</td>
			<td style="width:205px;">Advanced Cell Diagnostics&#160;</td>
			<td style="width:135px;">310010</td>
			<td style="width:188px;"></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">RNAscope Probe - Rn-Drd1a</td>
			<td>Advanced Cell Diagnostics&#160;</td>
			<td>317031</td>
			<td>Color channel 1, Green</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">RNAscope Probe - Rn-Th-C2</td>
			<td>Advanced Cell Diagnostics&#160;</td>
			<td>314651-C2</td>
			<td>Color channel 2, Orange</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">RNAscope Fluorescent Multiplex Reagent Kit</td>
			<td>Advanced Cell Diagnostics&#160;</td>
			<td>320850</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">ImmEdge Hydrophobic Barrier Pen</td>
			<td>Vector Laboratory</td>
			<td>H-4000</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">SuperFrost Plus Slides</td>
			<td>Fisher Scientific</td>
			<td>12-550-154%&#160;</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">4% paraformaldehyde</td>
			<td>Sigma-Aldrich</td>
			<td>158127-500G</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Sevoflurane</td>
			<td>Merritt Veterinary Supply</td>
			<td>347075</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Tissue-Tek vertical 24 slide rack</td>
			<td>Fisher Scientific</td>
			<td>NC9837976</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Tissue-Tek staining dish</td>
			<td>Fisher Scientific</td>
			<td>NC0731403</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Precision General Purpose Baths</td>
			<td>ThermoFisher Scientific</td>
			<td>TSGP28</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:329px;">Pretreatment 4</td>
			<td>Advanced Cell Diagnostics&#160;</td>
			<td>320850</td>
			<td>parts of kits</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:329px;">ProLong Gold anti-fade reagent</td>
			<td style="width:205px;">Life Technologies</td>
			<td style="width:135px;">P36930</td>
			<td style="width:188px;"></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:329px;">Amp 1-FL</td>
			<td>Advanced Cell Diagnostics&#160;</td>
			<td>320850</td>
			<td>parts of kits</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:329px;">Amp 2-FL</td>
			<td>Advanced Cell Diagnostics&#160;</td>
			<td>320850</td>
			<td>parts of kits</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:329px;">Amp 3-FL</td>
			<td>Advanced Cell Diagnostics&#160;</td>
			<td>320850</td>
			<td>parts of kits</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:329px;">Amp 4-FL-Alt A</td>
			<td>Advanced Cell Diagnostics&#160;</td>
			<td>320850</td>
			<td>parts of kits</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:329px;">EZ-C1 software package</td>
			<td style="width:205px;">Nikon Instruments</td>
			<td style="width:135px;"></td>
			<td>version 3.81b</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:329px;">SAS/STAT Software</td>
			<td style="width:205px;">SAS Institute, Inc.,</td>
			<td style="width:135px;"></td>
			<td>version 9.4</td>
		</tr>
	</tbody>
</table>

identification of dopamine d1-alpha receptor within rodent nucleus accumbens by an innovative rna in situ detection technology

Nel sistema nervoso centrale, il recettore di sottotipo D1-alfa (Drd1α) è il più abbondante recettore della dopamina (DA), che svolge un ruolo fondamentale nella regolazione dello sviluppo e crescita neuronale. Tuttavia, i meccanismi alla base di Drd1α anomalie di recettori che mediano le risposte comportamentali e modulazione della funzione di memoria di lavoro sono ancora poco chiari. Usando un romanzo RNA in situ ibridazione analisi, lo studio corrente ha identificato dopamina Drd1α del recettore e la tirosina idrossilasi (TH) espressione del RNA da correlati DA circuiti nella zona di nucleo accumbens (NAc) e regione di substantia nigra (SNR), rispettivamente. Drd1α espressione nel NAc Mostra un "discreto punto" modello di macchiatura. Sono state osservate differenze di sesso chiaro nell'espressione di Drd1α. Al contrario, TH Mostra un pattern di colorazione "cluster". Per quanto riguarda l'espressione TH, ratti femminili visualizzata un' più alta espressione di segnale per cella rispetto gli animali maschii. I metodi presentati qui forniscano una tecnica di romanzo in situ di ibridazione per indagare i cambiamenti nella disfunzione del sistema della dopamina durante la progressione di malattie del sistema nervoso centrale.

Lo studio corrente ha osservato un "punti discreti" metodo di colorazione per l'espressione di RNA nei recettori della dopamina D1-alfa (Drd1α) del NAc in ratti F344/N (Figura 1A). Segnali di fluorescenza individuali sono stati identificati facilmente e possono essere visto come unica "puntini", ognuno dei quali rappresenta una singola trascrizione di RNA all'interno della cellula. Per le immagini da NAc che visualizzano i "puntini discreti" modello di macch...

Watch this Scientific Journal Video about Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology at JoVE.com

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

Identificazione della dopamina D1-alfa del ricevitore all'interno del roditore Nucleus Accumbens da un innovativo RNA In Situ Detection Technology

Identificazione del recettore D1-alfa della dopamina nel nucleus accumbens è fondamentale per chiarire la disfunzione dei recettori D1 durante una malattia del sistema nervoso centrale. Abbiamo effettuato un romanzo determinazione del RNA in situ di ibridazione per visualizzare singole molecole di RNA in un'area specifica del cervello.

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in ...

The overall goal of this novel RNA in-situ hybridization assay is to visualize single RNA molecules in a specific brain area which are critical for clarifying dopaminergic dysfunction during central nervous system disease. This method can help answer key questions in the neuroscience field such as dopaminergic abnormalities mediating behavioral responses and modulating working memory function. The main advantage of this method is that it provides a novel in-situ hybridization technique to investigate dopamine system dysfunction during the progression of central nervous system diseases.Though this method can provide insight into overall expression levels of a target mRNA in a given brain region, it can also be applied to determine how mRNA transcripts are distributed throughout the cells of a given region. To begin this procedure, within five minutes after harvesting an adult rat brain, submerge it in liquid nitrogen for 15 seconds. Next, equilibrate the brain at minus 20 degrees Celsius in a cryostat for one hour.After an hour, cut 30 micron sections containing nucleus accumbens from 2.76 millimeters to 2.28 millimeters anterior to bregma and mount them onto the slides. Then dry the sections at minus 20 degrees Celsius for 10 minutes. Subsequently, immerse the slides in pre-chilled 4%paraformaldehyde for one hour at four degrees Celsius.Next, place the slides in an increasing ethanol gradient at room temperature then transfer the slides to fresh 100%ethanol for another five minutes. Afterward, place the slides on the absorbent paper to allow air dry. Draw a barrier around each section with a barrier pen.Let the barrier dry completely for one minute. To pre-treat the brain sections, turn on the oven and set the temperature to 40 degrees Celsius. Add three drops of pre-treatment reagent on each brain section.Then incubate the sections for 30 minutes at room temperature. Next, submerge the slides in 1X PBS for one minute at room temperature and transfer the slides to fresh 1X PBS for another minute. Slides should not stay in 1X PBS for longer than 15 minutes.To run the multiplex assay, warm the target probe for 10 minutes at 40 degrees Celsius in a water bath and then cool to room temperature. Place the RNA reagent at room temperature. Remove excess liquid from the slides with absorbent paper and place them back on the slide rack.Then add three drops of the Drd1alpha as probe on each sample slice and incubate the samples for two hours at 40 degrees Celsius. Afterward, wash the slides in 1X wash buffer for two minutes at room temperature twice. Then remove excess liquid from the slides with absorbent paper and place them back on the slide rack.Do not let brain sections dry out between incubation steps. Add three drops of AMP 1FL on each sample slice and incubate the samples for 30 minutes at 40 degrees Celsius. Afterward, wash the slides twice in 1X wash buffer for two minutes each time at room temperature.Then remove excess liquid from the slides with absorbent paper and place them back on the slide rack. Subsequently, add three drops of AMP 2FL on each sample slice. Incubate the samples for 15 minutes at 40 degrees Celsius.Next, submerge the slides in 1X wash buffer for two minutes at room temperature twice. Remove excess liquid from the slides with absorbent paper and place them back on the slide rack. Add three drops of AMP 3FL on each sample slice and incubate the samples for 30 minutes at 40 degrees Celsius.Next, wash the slides in 1X wash buffer for two minutes at room temperature twice. Remove excess liquid from the slides with absorbent paper and place them back on the slide rack. Then add three drops of AMP 4FL ALT A on each sample slice and incubate the samples for 15 minutes at 40 degrees Celsius.Wash the slide in 1X wash buffer for two minutes at room temperature twice. Remove excess liquid from the slides and immediately place two drops of mounting reagent onto each section. Next, place a 22 millimeter by 22 millimeter coverslip over each brain section.Then store the slides in the dark at two to eight degrees Celsius until dry. For imaging, turn on the confocal microscope and switch to a 60X objective. Obtain Z stack images and acquire images of Drd1alpha-labeled cells in the nucleus accumbens region.For images from the nucleus accumbens that displayed the discrete dots staining pattern, the dynamic range of expression was assessed using a semi-quantitative analysis. Each cell was scored from zero to nine based on the number of dots per cell. This hybridized signal score method can thus assess the extent and variability for Drd1alpha expression across individual cells.Here are the representative confocal images of Drd1alpha expression in male and female rats which have the discrete dots staining pattern. To account for the nested data structure, average count values were calculated for each category and used for statistical analysis. A Mann-Whitney U test was conducted to assess differences in median cell count in male and female rats.The frequency of cells in each category is analyzed using relative frequency and cumulative frequency. Female animals displayed a significant shift in the distribution with the greater frequency of cells in lower ordered categories relative to male animals. Overall, the results suggest clear sex differences in Drd1alpha expression in the nucleus accumbens.Once mastered, this technique can be done in seven hours if it is performed properly. While attempting this procedure, it's important to remember to work quickly as temperature changes or drying out of the tissue can damage the mRNA and hinder our ability to obtain a reliable signal. After watching this video, you should have a good understanding of how to perform the in-situ hybridization technique in cortical tissues, fluorescently image signal mRNA transcripts within chosen brain regions, and characterize the distribution of mRNA transcripts for statistical analysis.

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Neuroscience

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

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Currently, large-scale networks derived from dissociated neurons growing and developing in vitro on extracellular micro-transducer devices are the ...

Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression

Identificazione affidabile di vita neuroni dopaminergici in Culture Midbrain Utilizzando RNA Sequencing e TH-promotore-driven eGFP Expression

In Parkinson&#39;s Disease (PD) there is widespread neuronal loss throughout the brain with pronounced degeneration of dopaminergic neurons in the SNc, ...

Localization of Odorant Receptor Genes in Locust Antennae by RNA In Situ Hybridization

Localization of Odorant Receptor Genes in Locust Antennae by RNA In Situ Hybridization

Localizzazione dei geni del recettore degli odoranti nell&#39;antenna di Locust da RNA<em&gt; In Situ</em&gt; Ibridazione

Insects have evolved sophisticated olfactory reception systems to sense exogenous chemical signals. These chemical signals are transduced by Olfactory ...

Assessment of Dopaminergic Homeostasis in Mice by Use of High-performance Liquid Chromatography Analysis and Synaptosomal Dopamine Uptake

Valutazione dell'omeostasi dopaminergici in topi tramite l'uso di analisi di cromatografia liquida ad alte prestazioni e assorbimento della dopamina sinaptosomiali

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) ...