Expression of α2-adrenergic receptor subtypes in prenatal rat spinal cord

Yi Huang, W. Daniel Stamer, Todd L. Anthony, David V. Kumar, Paul A. St. John, John W. Regan

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

The results of molecular cloning have revealed three subtypes of the α2-adrenergic receptors (α2 AR) that have been defined α2C10 (α2A), α2C2 (α2B) and α2C4 (α2C). The differential expression of α2 AR subtypes is affected by developmental factors in rat submandibular gland, lung and brain. In the spinal cord of postnatal and adult rats, α2A and α2C AR subtypes are expressed and appear to mediate pain perception. However, the relative expression of α2 AR subtypes in the prenatal spinal cord is unknown. In the present study subtype-specific antibodies and reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the expression and localization of the α2 AR subtypes in sections of embryonic day 14 rat spinal cords and primary cultures of cells isolated from these cords. Spinal cords were removed from day 14 embryos, and were sectioned or used for the preparation of cell cultures. After 9 days in culture, neurons were examined by immunofluorescence microscopy or used for preparation of total RNA. In both intact spinal cords and isolated cells, positive immunoreactivity was detected with antibodies against α2A and α2B subtypes, but not with antibodies against the α2C subtype. Using a dual-labeling approach, anti-α2A and anti-α2B immunoreactivity was present on the same population of neurons. RT-PCR results were consistent with immunofluorescence studies, and showed that mRNA encoding the α2A and α2B subtypes was present in total RNA prepared from primary cultures of rat spinal cord neurons. In contrast to spinal cords of postnatal or adult rats that express α2A and α2C AR subtypes on different neurons, prenatal spinal cords contain α2A and α2B AR subtypes, and these two subtypes appear to be co-expressed in the same cells.

Original languageEnglish (US)
Pages (from-to)93-104
Number of pages12
JournalDevelopmental Brain Research
Volume133
Issue number2
DOIs
StatePublished - Feb 28 2002

Keywords

  • Adenylyl cyclase inhibition
  • Analgesia
  • Antibody
  • G-Protein coupled receptor
  • Immunohistochemistry
  • cAMP

ASJC Scopus subject areas

  • Developmental Neuroscience
  • Developmental Biology

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