Name: reliable identification of living dopaminergic neurons in midbrain cultures using rna sequencing and th-promoter-driven egfp expression
Uploaded: 2017-02-10T12:30:00
Description: Watch this Scientific Journal Video about Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression at JoVE.com

This work was supported by grants from the U.S. National Institutes of Health (DA017279, AG033954, DA037743), the Michael J. Fox Foundation, and the Caltech Innovation Initiative funding the Millard and Muriel Jacobs Genetics and Genomics Laboratory at the California Institute of Technology. We thank Brian Williams for optimising the RNA-Seq library protocol and for providing assistance. We thank Henry Amrhein for computational training. We thank Barbara J. Wold for the use of her equipment and laboratory space. We thank Igor Antoshechkin for library sequencing, and for facility management.

OBS: Alla experiment genomfördes i enlighet med de riktlinjer för skötsel och användning av djur som tillhandahålls av National Institutes of Health, och protokoll har godkänts av Institutional Animal Care och användning kommittén vid California Institute of Technology. 1. Härledning av primär Dopaminerga cellkulturer från embryonala Mouse Brain <ol><li> Lösningar och kultur Medium <ol><li> Framställning av askorbinsyra Stock Solution <ol><li> Väg upp 352 mg askorbinsyra. Til...

<ol>
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Här isolera vi enskilda celler från en heterogen population med hjälp av en fluorescerande tagg, sedan studera varje cell med encelliga RNA-punkter. Vi rapporterar att 100% av de levande TH-EGFP celler som vi skördade och sekvense var verkligen dopaminerga neuroner, baserat på förekomsten av följande tre DA-relaterade gentranskript, TH, DDC och slc6a3. Alla TH-EGFP positiva celler uttryckte dessa tre gener som bedöms av RNA-Seq. Detta var samstämmiga med elektrofysiologiska studier som visade att varje TH-EGFP ...

Parkinsons sjukdom (PD) är en obotlig, åldersrelaterad neurodegenerativ sjukdom. Orsaken till denna relativt vanlig sjukdom fortfarande dåligt förstådd. Det finns en utbredd neuronal förlust i hela hjärnan, med uttalad neuronal degenerering av dopaminerga neuroner i substantia nigra (SNC), vilket leder till diagnostiska kliniska tecken på bradykinesi, stelhet och skakningar. Primära blandade kulturer innehållande SNC dopaminerga nervceller är särskilt relevanta för Parkinsons sjukdom. Ventrala tegmentområdet (VTA) dopaminerga neuroner har varit inblandade i belöning och missbruk. Ventral mesencefala (VM) primära blandade embryonala kulturer innehåller både SNC och VTA dopaminerga (DA) neuron och GABAergic nervceller. VM primära kulturer kan vara användbara för neuroprotektion analyser och för att belysa den selektiva sårbarhet av dopaminerga neuroner. Det finns ingen tillförlitlig sätt att identifiera dopaminerga celler i odling baserat på morfologi. hanre vi utveckla teknik för att identifiera och skörda enstaka dopaminerga neuroner, och bygga encelliga, högavkastande RNA sekvense bibliotek. Vi rapporterar representativa RNA transkriptom data för enkel dopaminerga och GABAergic nervceller isolerade från mitthjärnan kulturer. Protokollet kan användas för neuroprotektion, neurodegeneration, och farmakologiska analyser för att studera effekterna av olika behandlingar på DA / GABA transkriptom. Eftersom dopaminerga neuroner utgör en liten minoritet av de nervceller som uttrycks i primär VM kulturer, kommer förmågan att på ett tillförlitligt sätt identifiera dessa nervceller i levande kulturer möjliggöra ett utökat sortiment av single-cellstudier. Dessa nya tekniker kommer att underlätta framsteg inom förstå de mekanismer som äger rum på cellnivå och kan ha applikationer på andra ställen i områdena neurovetenskap och molekylärbiologi.

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		<tr height="40">
			<td height="40" style="height:40px;width:395px;">Papain</td>
			<td style="width:241px;">Worthington Biochemical Corporation&#160;</td>
			<td style="width:108px;">LS003126</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Hanks&#8217; Balanced Salt Solution (HBSS), 1X</td>
			<td style="width:241px;">Gibco&#160;</td>
			<td style="width:108px;">14175-095</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Donor Equine Serum</td>
			<td style="width:241px;">Thermo Scientific&#160;</td>
			<td style="width:108px;">SH30074.03</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Bovine Serum Albumin (BSA)</td>
			<td style="width:241px;">Sigma&#160;</td>
			<td style="width:108px;">A7030-50G</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Medium: Neurobasal medium</td>
			<td style="width:241px;">Gibco&#160;</td>
			<td style="width:108px;">21103-049</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:395px;">Culture media that contains a stabilized form of L-glutamine, L-alanyl-L-glutamine: GlutaMAX</td>
			<td style="width:241px;">Gibco&#160;</td>
			<td style="width:108px;">35050-061</td>
			<td style="width:177px;">0.5 mM final concentration.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">L-Ascorbic acid</td>
			<td style="width:241px;">Sigma&#160;</td>
			<td style="width:108px;">A7506</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:395px;">B27</td>
			<td style="width:241px;">Gibco&#160;</td>
			<td style="width:108px;">17504-044</td>
			<td style="width:177px;">50 X stock solution, 1 X final concentration.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">35 mm glass bottom dishes</td>
			<td style="width:241px;">MatTek&#160;&#160;</td>
			<td style="width:108px;">P35GC-1.5-10-C</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Poly-L-Ornithine</td>
			<td style="width:241px;">Sigma&#160;</td>
			<td style="width:108px;">P4957</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:395px;">Laminin</td>
			<td style="width:241px;">Sigma&#160;</td>
			<td style="width:108px;">L2020</td>
			<td style="width:177px;">Stored at -20&#176;C in 20 &#181;L aliquots</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Deoxyribonuclease I (DNase)</td>
			<td style="width:241px;">Sigma&#160;</td>
			<td style="width:108px;">DN25</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Blue pipette tips</td>
			<td style="width:241px;">Sorenson Bioscience,</td>
			<td style="width:108px;">&#160;Inc. 10130</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">P1000</td>
			<td style="width:241px;"></td>
			<td style="width:108px;"></td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">10 mm shallow Petri dish</td>
			<td style="width:241px;">VWR&#160;</td>
			<td style="width:108px;">25384-324</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">37&#176;C Water bath</td>
			<td style="width:241px;"></td>
			<td style="width:108px;"></td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">37&#176;C, 5% humidity incubator</td>
			<td style="width:241px;"></td>
			<td style="width:108px;"></td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">16% paraformaldehyde (PFA)</td>
			<td style="width:241px;">Electron Microscopy Sciences&#160;</td>
			<td style="width:108px;">15710</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Triton X-100</td>
			<td style="width:241px;">Sigma</td>
			<td style="width:108px;">&#160;X100-500ML</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:395px;">Donkey serum</td>
			<td style="width:241px;">Equitech&#160;</td>
			<td style="width:108px;">SD-0100HI</td>
			<td style="width:177px;">100% stock solution, 1% final concentration.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Standard Pattern surgical scissors</td>
			<td style="width:241px;">Fine Science Tools</td>
			<td style="width:108px;">14000-14</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Forceps - 2x3 teeth</td>
			<td style="width:241px;">Fine Science Tools</td>
			<td style="width:108px;">11022-14</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Forceps - Dumont #55</td>
			<td style="width:241px;">Fine Science Tools</td>
			<td style="width:108px;">11295-51</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Forceps - Dumont #5</td>
			<td style="width:241px;">Fine Science Tools</td>
			<td style="width:108px;">11252-23</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">10X PBS, pH 7.4</td>
			<td style="width:241px;">Gibco&#160;</td>
			<td style="width:108px;">70011-044</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Dulbecco&#39;s Phosphate Buffered solution (DPBS) 1X</td>
			<td style="width:241px;">Gibco</td>
			<td style="width:108px;">14190-144</td>
			<td style="width:177px;">500 ml&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">21 guage needle&#160;</td>
			<td style="width:241px;">BD PrecisionGlide Needle</td>
			<td style="width:108px;">305167</td>
			<td style="width:177px;">100 needles</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Micropipette puller</td>
			<td style="width:241px;">Sutter Instrument.</td>
			<td style="width:108px;">model P-87</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Micromanipulator&#160;</td>
			<td style="width:241px;">Sutter Instrument</td>
			<td style="width:108px;">&#160;MP-285</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:395px;">Sequencing Kit: SMARTer Ultra Low RNA Kit for Illumina Sequencing</td>
			<td style="width:241px;">Clontech</td>
			<td style="width:108px;">634936</td>
			<td style="width:177px;">100rnxs,incl Advantage2PCR Kit</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:395px;">oligonucleotide (12 &#181;M): SMARTer IIA oligonucleotide (12 &#181;M)</td>
			<td style="width:241px;"></td>
			<td style="width:108px;"></td>
			<td style="width:177px;">From the SMARTer Kit&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Reverse transcriptase (100 units)</td>
			<td style="width:241px;">SMARTcribe reverse transcriptase</td>
			<td style="width:108px;"></td>
			<td style="width:177px;">From the SMARTer Kit&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Primer 1</td>
			<td style="width:241px;">3&#8217; CDSIIa primer&#160;</td>
			<td style="width:108px;"></td>
			<td style="width:177px;">From the SMARTer Kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Primer 2</td>
			<td style="width:241px;">IS PCR primer</td>
			<td style="width:108px;"></td>
			<td style="width:177px;">From the SMARTer Kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">50X 2 polymerase mix</td>
			<td style="width:241px;">50X Advantage 2 polymerase mix</td>
			<td style="width:108px;"></td>
			<td style="width:177px;">From the SMARTer Kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">10X PCR buffer</td>
			<td style="width:241px;">10X Advantage 2 PCR buffer</td>
			<td style="width:108px;"></td>
			<td style="width:177px;">From the SMARTer Kit</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">RNA Spikes</td>
			<td style="width:241px;">ThermoFisher</td>
			<td style="width:108px;">4456740</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">PCR cooler rack</td>
			<td style="width:241px;">IsoFreeze PCR cooler rack.</td>
			<td style="width:108px;"></td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">DNA Sample Preparation Kit&#160;</td>
			<td style="width:241px;">(Illumina/Nextera) Nexter</td>
			<td style="width:108px;">FC-121-1030</td>
			<td style="width:177px;">24 Samples</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">PCR master mix</td>
			<td style="width:241px;">Nextera PCR master mix (NPM)</td>
			<td style="width:108px;"></td>
			<td style="width:177px;">From the Nextera Kit&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">PCR primer cocktail (PPC)&#160;</td>
			<td style="width:241px;">Nextera PCR primer cocktail (PPC)&#160;</td>
			<td style="width:108px;"></td>
			<td style="width:177px;">From the Nextera Kit</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Fluorometer</td>
			<td style="width:241px;">The Qubit ThermoFisher</td>
			<td style="width:108px;">Q33216</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">HS DNA BioAnalyzer kit</td>
			<td style="width:241px;">Agilent</td>
			<td style="width:108px;">5067-4626</td>
			<td style="width:177px;">110rxns</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:395px;">Electrophoresis system&#160;</td>
			<td style="width:241px;">Agilent 2100 Bioanalyzer.</td>
			<td style="width:108px;">G2938C</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Magnetic beads: Agencourt AMPure&#160;</td>
			<td style="width:241px;">Beckman Coulter</td>
			<td style="width:108px;">A63880</td>
			<td style="width:177px;">5ml</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">RNAse wipe: RNAse Zap wipes</td>
			<td style="width:241px;">Ambion</td>
			<td style="width:108px;">AM9786</td>
			<td style="width:177px;">Size 100 Sheets</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Qubit ds HS Assay Kit</td>
			<td style="width:241px;">Molecular probes&#160;</td>
			<td style="width:108px;">Q32854</td>
			<td style="width:177px;">500 assays</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Gel extraction kit: Qiaquick Gel Extraction Kit</td>
			<td style="width:241px;">Qiagen&#160;</td>
			<td style="width:108px;">28704</td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Glass capillary tubing</td>
			<td style="width:241px;">(Kimax-51, 1.5&#8211;1.8 mm o.d.)</td>
			<td style="width:108px;"></td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Microforge</td>
			<td style="width:241px;">Narishige (or equivalent)&#160;</td>
			<td style="width:108px;"></td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:395px;">Sequencer</td>
			<td style="width:241px;">Illumina HiSeq instrument</td>
			<td style="width:108px;"></td>
			<td style="width:177px;"></td>
		</tr>
		<tr height="80">
			<td height="80" style="height:80px;width:395px;">GENSAT tyrosine hydroxylase-eGFP mouse strain&#160;</td>
			<td style="width:241px;"></td>
			<td style="width:108px;">MMRRC stock number: 000292-UNC</td>
			<td style="width:177px;">Stock Tg(Th-EGFP)DJ76Gsat/Mutant Mouse Regional Research Center</td>
		</tr>
	</tbody>
</table>

reliable identification of living dopaminergic neurons in midbrain cultures using rna sequencing and th-promoter-driven egfp expression

<html> Vid Parkinsons sjukdom (PD) det finns en utbredd neuronal förlust i hela hjärnan med uttalad degeneration av dopaminerga neuroner i SNC, vilket leder till bradykinesi, stelhet och skakningar. Identifieringen av levande dopaminerga neuroner i primära Ventral mesencefala (VM) kulturer med hjälp av en fluorescerande markör ger ett alternativt sätt att studera den selektiva sårbarhet dessa nervceller utan att förlita sig på immunfärgning av fixerade celler. Här, vi isolera, dissociera och kultur mus VM neuroner för 3 veckor. Vi identifierar sedan dopaminerga neuroner i kulturer med hjälp av EGFP fluorescens (drivs av en tyrosinhydroxylas (TH) promotor). Enskilda nervceller skördas i mikrocentrifugrör med hjälp av glasmikropipetter. Nästa, vi lysera de skördade cellerna och utföra cDNA-syntes och transposon-medierad &quot;tagmentation&quot; för att producera enkelcells RNA-Seq bibliotek 1, 2,ass = &quot;xref&quot;&gt; 3, 4, 5. Efter att ha passerat genom kvalitetskontroll check, är biblioteken encelliga sekvenserades och efterföljande analys utförs för att mäta genuttryck. Vi rapporterar transkriptom resultat för enskilda dopaminerga och GABAergic nervceller isolerade från mitthjärnan kulturer. Vi rapporterar att 100% av de levande TH-EGFP celler som skördades och sekvenserades var dopaminerga neuroner. Dessa tekniker kommer att ha omfattande tillämpningar inom neurovetenskap och molekylärbiologi.

Här rapporterar vi ett protokoll för odling ventrala mitthjärnan nervceller från en mus som uttrycker EGFP drivs av en tyrosinhydroxylas promotorsekvens, skörda enskilda fluorescerande nervceller från kulturerna, och mätning av deras RNA transkriptom med hjälp av RNA-punkter (Figur 1). Data visade att 100% av de TH-EGFP-positiva celler som skördades och sekvenser var dopaminerga neuroner, baserat på förekomsten av följande tre DA-relaterade gentranskript, TH,...

Watch this Scientific Journal Video about Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression at JoVE.com

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

Vid Parkinsons sjukdom (PD), substantia nigra (SNC) dopaminerga neuroner degenererar, vilket leder till motorisk dysfunktion. Här rapporterar vi ett protokoll för odling ventrala mitthjärnan nervceller från en mus som uttrycker EGFP drivs av en tyrosinhydroxylas (TH) promotorsekvens, skörda enskilda fluorescerande nervceller från kulturerna, och mätning av deras transkriptom med hjälp av RNA-punkter.

Tillförlitlig identifiering av Living dopaminerga neuroner i mellanhjärnan kulturer Använda RNA sekvensering och TH-promotor-driven EGFP uttryck

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

Research

JoVE Journal

Neuroscience

Application of a NMDA Receptor Conductance in Rat Midbrain Dopaminergic Neurons Using the Dynamic Clamp Technique

Tillämpning av en NMDA-receptorn Konduktans av dopaminerg Rat mitthjärnan Nervceller med det dynamiska Clamp Teknik

Neuroscientists study the function of the brain by investigating how neurons in the brain communicate. Many investigators look at changes in the ...

Organotypic Slice Cultures of Embryonic Ventral Midbrain: A System to Study Dopaminergic Neuronal Development in vitro

Organotypic Slice Cultures of Embryonic Ventral Midbrain: A System to Study Dopaminergic Neuronal Development in vitro

Organotypic Slice kulturer av embryonala på magen mitthjärnan: Ett system att studera Dopaminerga nervsystemets utveckling In vitro</em

The  mouse is an excellent model organism to study mammalian brain development due  to the abundance of molecular and genetic data. However, the ...

In ovo Electroporation in Chick Midbrain for Studying Gene Function in Dopaminergic Neuron Development

In ovo Electroporation in Chick Midbrain for Studying Gene Function in Dopaminergic Neuron Development

I ovo elektroporering i Chick mitthjärnan för att studera genfunktion av dopaminerg Neuron utveckling

Dopaminergic neurons located in the ventral midbrain control movement, emotional behavior, and reward mechanisms1-3. The dysfunction of ventral midbrain ...

In ovo Expression of MicroRNA in Ventral Chick Midbrain

In ovo Expression of MicroRNA in Ventral Chick Midbrain

I ovo Uttryck av MikroRNA i ventrala Chick mitthjärnan

Non-coding RNAs are additional players in regulating gene expression. Targeted in ovo electroporation of specific areas provides a unique tool for spatial ...

Assessing Neurodegenerative Phenotypes in Drosophila Dopaminergic Neurons by Climbing Assays and Whole Brain Immunostaining

Assessing Neurodegenerative Phenotypes in Drosophila Dopaminergic Neurons by Climbing Assays and Whole Brain Immunostaining

Uppskatta Neurodegenerativa Fenotyper i<em&gt; Drosophila</em&gt; Dopaminerga neuroner genom att klättra Analyser och Helhjärnstänkande immunostaining

Drosophila melanogaster is a valuable model organism to study aging and pathological degenerative processes in the nervous system. The advantages of the ...

Primary Culture of Mouse Dopaminergic Neurons

Primär kultur av mus dopaminerga neuroner

Dopaminergic neurons represent less than 1% of the total number of neurons in the brain. This low amount of neurons regulates important brain functions ...

Environmental Modulations of the Number of Midbrain Dopamine Neurons in Adult Mice

Miljö modulation av antalet mitthjärnan dopaminneuroner i vuxna möss

Long-lasting changes in the brain or &#8216;brain plasticity&#8217; underlie adaptive behavior and brain repair following disease or injury. Furthermore, ...

Isolation, Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains

Isolering, kultur och långsiktigt underhåll av primär mesencefala dopaminerga neuron från embryonala Gnagare Brains

Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson&#8217;s diseae. Study of the biological processes ...

External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures

Extern Excite av nervceller Använda elektriska och magnetiska fält i en-och två-dimensionella kulturer

A neuron will fire an action potential when its membrane potential exceeds a certain threshold. In typical activity of the brain, this occurs as a result ...

Determining Cell-surface Expression and Endocytic Rate of Proteins in Primary Astrocyte Cultures Using Biotinylation

Bestämning av cellytans uttryck och endocytisk hastighet för proteiner i primära astrocytkulturer med användning av biotinylering

Cell-surface proteins mediate a wide array of functions. In many cases, their activity is regulated by endocytic processes that modulate their levels at ...