Basic Information
-
Description
-
- G protein-coupled receptors (GPCRs), also designated seven transmembrane (7TM) receptors and heptahelical receptors, are a protein family which interact with G proteins (heterotrimeric GTPases) to synthesize intracellular second messengers such as diacylglycerol, cyclic AMP, inositol phosphates and calcium ions. Their diverse biological functions range from vision and olfaction to neuronal and endocrine signaling and are involved in many pathological conditions.
- G protein receptor 84 (GPR84), a member of the GCPR 1 family, is an orphan GCPR expressed in bone marrow, brain, heart, muscle, colon, thymus, spleen, kidney, liver, placenta, intestine, lung and peripheral blood leukocytes. In activated T cells, GPR84 regulates early interleukin-4 (IL-4) gene expression.
-
Targeting strategy
-
Gene targeting strategy for B-hGPR84 mice.
The exon 2 of mouse Gpr84 gene that encodes the full-length protein was replaced by human GPR84 exon 2 in B-hGPR84 mice.
-
mRNA expression analysis
-
Strain specific analysis of GPR84 mRNA expression in wild-type C57BL/6N mice, B-hGPR84 mice and B-GPR84 KO mice by RT-PCR. Brain RNA were isolated from wild-type C57BL/6N mice (+/+), homozygous B-hGPR84 mice (H/H) and B-GPR84 KO mice (Δ/Δ), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human GPR84 primers. Mouse Gpr84 mRNA was detectable only in wild-type C57BL/6N mice. Human GPR84 mRNA was detectable only in homozygous B-hGPR84 mice.
-
Protein expression analysis
-
Western blot analysis of GPR84 protein expression in homozygous B-hGPR84 mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hGPR84 mice (H/H), and then analyzed by western blot with anti-GPR84 antibody. (anti-human GPR84 antibody: Santa Cruz, sc-293447). 40 μg total proteins were loaded for western blotting analysis. GPR84 was detected in lung, liver, heart and bone marrow, as the antibody is cross-recognized between human and mouse GPR84. (There was no β-actin in the heart tissue, there was no GAPDH in the bone marrow tissue.)
