Molecular cloning, sequencing, and distribution of feline GnRH receptor (GnRHR) and resequencing of canine GnRHR

• Published in: Theriogenology 2015-01, Vol.83 (2), p.266-275
• Main Authors: Samoylov, Alexandre M., Napier, India D., Morrison, Nancy E., Martin, Douglas R., Cox, Nancy R., Samoylova, Tatiana I.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animal reproduction; Animals; Base Sequence; Cats - genetics; Cloning, Molecular; Contraception; DNA, Complementary - chemistry; DNA, Complementary - genetics; Dogs - genetics; Domestic cat; Female; Gene expression; GnRH receptor; Humans; Male; Mice; Organ Specificity; Pituitary; Pituitary Gland - chemistry; Receptors, LHRH - analysis; Receptors, LHRH - chemistry; Receptors, LHRH - genetics; Reverse Transcriptase Polymerase Chain Reaction - veterinary; RNA, Messenger - analysis; Sequence Analysis - veterinary; Sequence Homology
• ISSN: 0093-691X; 1879-3231
• DOI: 10.1016/j.theriogenology.2014.09.020

GnRH receptors play vital roles in mammalian reproduction via regulation of gonadotropin secretion, which is essential for gametogenesis and production of gonadal steroids. GnRH receptors for more than 20 mammalian species have been sequenced, including human, mouse, and dog. This study reports the molecular cloning and sequencing of GnRH receptor (GnRHR) cDNA from the pituitary gland of the domestic cat, an important species in biomedical research. Feline GnRHR cDNA is composed of 981 nucleotides and encodes a 327 amino acid protein. Unlike the majority of mammalian species sequenced so far, but similar to canine GnRHR, feline GnRHR protein lacks asparagine in position three of the extracellular domain of the protein. At the amino acid level, feline GnRHR exhibits 95.1% identity with canine, 93.8% with human, and 88.9% with mouse GnRHR. Comparative sequence analysis of GnRHRs for multiple mammalian species led to resequencing of canine GnRHR, which differed from that previously published by a single base change that translates to a different amino acid in position 193. This single base change was confirmed in dogs of multiple breeds. Reverse transcriptase PCR analysis of GnRHR messenger RNA in different tissues from four normal cats indicated the presence of amplicons of varying lengths, including full-length as well as shortened GnRHR amplicons, pointing to the existence of truncated GnRHR transcripts in the domestic cat. This study is the first insight into molecular composition and expression of feline GnRHR and promotes better understanding of receptor organization, and distribution in various tissues of this species.