Document Type

Honors Project

First Advisor

Dr. Robyn Puffenbarger

Degree Award Date

Spring 2004

Keywords

Transcriptional Regulation, CB2 Receptor, Positional Sub-Cloning, Promoter Target Studies

Disciplines

Biology | Microbiology

Abstract

The cannabinoid receptor system has many physiological roles including altering mood, appetite, locomotion, pain perception, inflammation, and vasodilation. The primary cannabinoid receptor of the immune system is CB2, which can be found on T cells, macrophages, and B cells. In macrophages, CB2 expression varies throughout different stages of an immune response, from low expression in resident cells, to higher levels in responsive cells, and back to low expression in activated cells. Therefore, understanding the regulation of CB2 expression is important for determining how and when immune cells are receptive to cannabinoid ligand modulation. Our primary goal is to define the promoter elements responsible for CB2 expression in immune cells. To characterize the genetic regulation of CB2, a portion of mouse chromosome 4 containing the previously defined CB2 exons (exon-lA, exon-lB, exon-2) was acquired. Various deletion constructs of three putative promoter sites have been made via PCR. To elucidate the activity of the CB2 promoter, the constructs were cloned into a luciferase reporter system. Macrophage cell lines RA W264.7 and P388D1 were used to characterize the genetic regulation of CB2. An 849 bp construct upstream of exon-lB has shown increased luciferase activity compared to a 239 bp in the same region and a 707 bp construct upstream of exon-lA. Protein assays have indicated an equal protein concentration of each sample tested suggesting a possible promoter region upstream of exon-lB.

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