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Real-Time Quantitative Reverse Transcription PCR for Detection of Opioid Receptors in Immune Cells.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2201, pp. 83-95. - Publication Year :
- 2021
-
Abstract
- Real-time quantitative reverse transcription-PCR (qRT-PCR ) is a highly sensitive molecular biology method based on the amplification of the cDNA of mRNA to detect and quantify the levels of mRNA of interest. In this chapter, we describe real-time qRT-PCR to detect and quantify mRNA of opioid receptors in immune cells. Specifically, we analyze mouse immune cells isolated from the blood and sciatic nerves exposed to a chronic constriction injury, which represents a model of neuropathic pain. We describe in detail the requirements and techniques to induce the chronic constriction injury, to isolate immune cells from the blood and injured nerves, to isolate the total RNA from immune cells, to perform a cDNA reverse transcription from the total RNA, and to perform real-time qRT-PCR for μ-, δ-, and κ-opioid receptor mRNAs.
- Subjects :
- Animals
Humans
Male
Mice
Mice, Inbred C57BL
Neuralgia
RNA, Messenger genetics
Receptors, G-Protein-Coupled genetics
Receptors, G-Protein-Coupled metabolism
Receptors, Opioid genetics
Receptors, Opioid immunology
Receptors, Opioid metabolism
Receptors, Opioid, delta genetics
Receptors, Opioid, kappa genetics
Receptors, Opioid, mu immunology
Receptors, Opioid, mu metabolism
Reverse Transcription
Sciatic Nerve injuries
Real-Time Polymerase Chain Reaction methods
Receptors, Opioid, mu genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 2201
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 32975791
- Full Text :
- https://doi.org/10.1007/978-1-0716-0884-5_8