The murine cysteinyl leukotriene 2 (CysLT2) receptor. cDNA and genomic cloning, alternative splicing, and in vitro characterization.
Two classes of cysteinyl leukotriene receptor, CysLT(1) and CysLT(2), have been identified and pharmacologically characterized in human tissues. Although the CysLT(1) receptor mediates the proinflammatory effects of leukotrienes in human asthma, the physiological roles of CysLT(2) receptor are not defined, and a suitable mouse model would be useful in delineating function. We report here the molecular cloning and characterization of the mouse CysLT(2) receptor (mCysLT(2)R) from heart tissue. mCysLT(2)R cDNA encodes a protein of 309 amino acids, truncated at both ends compared with the human ortholog (hCysLT(2)R). The gene resides on the central region of mouse chromosome 14 and is composed of 6 exons with the entire coding region located in the last exon. Two 5'-untranslated region splice variants were identified with the short form lacking exon 3 as the predominant transcript. Although the overall expression of mCysLT(2)R is very low, the highest expression was detected in spleen, thymus, and adrenal gland by ribonuclease protection assay, and discrete sites of expression in heart were observed by in situ hybridization. Intracellular calcium mobilization in response to cysteinyl leukotriene administration was detected in human embryonic kidney 293T cells transfected with recombinant mCysLT(2)R with a rank order of potency leukotriene C(4)(LTC(4) ) = LTD(4)>>LTE(4). [(3)H]LTD(4) binding to membranes expressing mCysLT(2)R could be effectively competed by LTC(4) and LTD(4) and only partially inhibited by LTE(4) and BAYu9773. The identification of mCysLT(2)R will be useful for establishing CysLT(2)R-deficient mice and determining novel leukotriene functions.
Hui
Y
Y
Center for Experimental Therapeutics, Department of Pharmacology, University of Pennsylvania, Philadelphia, PA 19104, USA.
Yang
G
G
Galczenski
H
H
Figueroa
D J
DJ
Austin
C P
CP
Copeland
N G
NG
Gilbert
D J
DJ
Jenkins
N A
NA
Funk
C D
CD
D020121:5' Untranslated Regions; D000311:Adrenal Glands / Q000378:metabolism; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D015152:Blotting, Northern; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003433:Crosses, Genetic; D018076:DNA, Complementary / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005091:Exons; D006801:Humans; D017403:In Situ Hybridization; D007438:Introns; D017997:Leukotriene C4 / Q000378:metabolism; D017998:Leukotriene D4 / Q000378:metabolism; D008565:Membrane Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D011869:Radioligand Assay; D018077:Receptors, Leukotriene / Q000737:chemistry* / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism; D014018:Tissue Distribution; D014162:Transfection
D020121:5' Untranslated Regions; D018076:DNA, Complementary; D008565:Membrane Proteins; D012333:RNA, Messenger; D018077:Receptors, Leukotriene; D011994:Recombinant Proteins; D017997:Leukotriene C4; D017998:Leukotriene D4; C412375:cysteinyl leukotriene receptor 2; D002118:Calcium