title: Identification of mRNA for endocannabinoid biosynthetic enzymes within hippocampal pyramidal cells and CA1 stratum radiatum interneuron subtypes using quantitative real-time polymerase chain reaction.
abstract: The hippocampus is required for short-term memory and contains both excitatory pyramidal cells and inhibitory interneurons. These cells exhibit various forms of synaptic plasticity, the mechanism underlying learning and memory. More recently, endocannabinoids were identified to be involved in synaptic plasticity. Our goal was to describe the distribution of endocannabinoid biosynthetic enzymes within CA1 stratum radiatum interneurons and CA3/CA1 pyramidal cells. We extracted mRNA from single interneurons and pyramidal cells and used real-time quantitative polymerase chain reaction (RT-PCR) to detect the presence of 12-lipoxygenase, N-acyl-phosphatidylethanolamine-specific phospholipase D, diacylglycerol lipase α, and type I metabotropic glutamate receptors, all known to be involved in endocannabinoid production and plasticity. We observed that the expression of endocannabinoid biosynthetic enzyme mRNA does occur within interneurons and that it is coexpressed with type I metabotropic glutamate receptors, suggesting interneurons have the potential to produce endocannabinoids. We also identified that CA3 and CA1 pyramidal cells express endocannabinoid biosynthetic enzyme mRNA. Our data provide the first molecular biological evidence for putative endocannabinoid production in interneurons, suggesting their potential ability to regulate endocannabinoid-mediated processes, such as synaptic plasticity.
authors.affiliation: Brigham Young University, Department of Physiology and Developmental Biology, Provo, UT 84602, USA.
authors.forename: C B
authors.initials: CB
authors.lastname: Merrill
authors.forename: M
authors.initials: M
authors.lastname: McNeil
authors.forename: R C
authors.initials: RC
authors.lastname: Williamson
authors.forename: B R
authors.initials: BR
authors.lastname: Poole
authors.forename: B
authors.initials: B
authors.lastname: Nelson
authors.forename: S
authors.initials: S
authors.lastname: Sudweeks
authors.forename: J G
authors.initials: JG
authors.lastname: Edwards
mesh_terms: D000818:Animals; D056547:CA1 Region, Hippocampal / Q000201:enzymology*; D056654:CA3 Region, Hippocampal / Q000378:metabolism; D063388:Endocannabinoids / Q000096:biosynthesis*; D007395:Interneurons / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D017966:Pyramidal Cells / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D063388:Endocannabinoids; D012333:RNA, Messenger