Detection of opioid receptor mRNA by RT-PCR reveals alternative splicing for the delta- and kappa-opioid receptors.
The three mu-, delta- and kappa-opioid receptors have recently been cloned and characterized at the molecular level. Our analysis of opioid receptor transcripts by RT-PCR revealed two PCR products derived from delta and kappa mRNAs with size higher than expected from the known cDNA sequences. DNA sequencing showed additional nucleotides inserted between the known splice sites, indicating the possible existence of alternative splicing pathways for delta and kappa receptors. The novel delta-opioid receptor transcript is expressed in mouse brain and contains a 243 bp insertion. This additional sequence is located at the splice junction between the first and second coding exons and is encoded by a single exon located 9 kb upstream exon 2 in the mDOR gene. The other alternative transcript occurs in human monocytic and T lymphocytic cell lines and encodes a novel form of the kappa-opioid receptor. The PCR product presents a 23 bp deletion at the 3' end of exon 2 followed by a 246 bp insertion found between exons 2 and 3. In the hKOR gene, this insertion is encoded by two DNA segments. One of them is located 0.4 kb downstream exon 2 while the second is flanking exon 3 on the 5' side. Both novel putative delta and kappa exons present in-frame stop codons that would lead to truncated receptor proteins. A possible functional or regulatory role of these shorter proteins in opioid function remains to be determined.
Gavériaux-Ruff
C
C
CNRS UPR 9050 Laboratoire des Récepteurs et Protéines Membranaires, ESBS, Illkirch, France. gaveriau@esbs.u-strasbg.fr
Peluso
J
J
Befort
K
K
Simonin
F
F
Zilliox
C
C
Kieffer
B L
BL
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D018076:DNA, Complementary / Q000302:isolation & purification; D005091:Exons; D016678:Genome; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D017465:Receptors, Opioid, delta / Q000235:genetics*; D017473:Receptors, Opioid, kappa / Q000235:genetics*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D012333:RNA, Messenger; D017465:Receptors, Opioid, delta; D017473:Receptors, Opioid, kappa