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Profiling Voltage-gated Potassium Channel mRNA Expression in Nigral Neurons using Single-cell RT-PCR Techniques
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JoVE Journal Neuroscience
Profiling Voltage-gated Potassium Channel mRNA Expression in Nigral Neurons using Single-cell RT-PCR Techniques

Profiling Voltage-gated Potassium Channel mRNA Expression in Nigral Neurons using Single-cell RT-PCR Techniques

DOI:
10.3791/3136-v

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07:31 min

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September 27, 2011

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,
1Department of Pharmacology,University of Tennessee College of Medicine

Chapters

  • 00:05Title
  • 00:54Brain Slice Preparation
  • 01:42Electrophysiological Fingerprinting of Nigral Neurons
  • 02:48Cytoplasm Aspiration
  • 03:29Reverse Transcription (RT) Reaction
  • 04:23Two Stage PCR Amplification
  • 05:14Electrophysiology and Gene Expression in Selected Neurons
  • 07:04Conclusion

Summary

Automatic Translation

Automatic Translation

Neurons are first characterized electrophysiologically. Then the cytoplasm from the recorded neuron is aspirated and subjected to reverse transcription-PCR analysis to detect the expression of mRNAs for neurotransmitter synthesis enzymes, ion channels, and receptors.

Tags

ProfilingVoltage-gated Potassium ChannelMRNA ExpressionNigral NeuronsSingle-cell RT-PCR TechniquesMammalian Central Nervous SystemNeuronsMolecular CharacteristicsFunctional CharacteristicsMorphologyGene Expression AnalysisSpecific Neuron TypeWhole-cell Patch Clamp Recording TechniquesSingle-cell Reverse Transcription Polymerase Chain Reaction (scRT-PCR)MRNA Expression ProfilingSubstantial NigraElectrophysiological TechniquesNeurophysiological PropertiesFunctional PropertiesCytoplasm AspirationScRT-PCR AnalysisNeurotransmitter Synthesis EnzymesReceptorsIon ChannelsSelectivitySensitivityImmunohistochemistryAntibody SuitabilityProtein Expression Level

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