Cloning and characterization of the ionotropic GABA receptor subunit ρ1 from pig (Sus scrofa).
Since human and pig eyes have remarkably anatomical and physiological similitudes swine models have been broadly used for functional studies and therapeutic research. Recently, a GABAρ-mediated relaxation of retinal vascularity suggested that GABAρ signaling may be used to improve retinal blood flow in vascular-driven impaired vision, and a further molecular characterization of GABAρ receptors would be beneficial. However, none of the GABAρ type subunits from pigs has been yet cloned; Among the 19 subunits that compose the family of GABAA receptors, ρ1-3 subunits are capable of forming homomeric channels. These homomeric receptors are particularly interesting because their pharmacological and kinetic properties are notably different from receptors composed by other GABAA subunits. Here we report the cloning of the GABAρ1subunit from the pig and the functional expression of homomeric channels in Xenopus oocytes. The most notable difference found in the pig GABAρ1 receptor was the absence of a stretch of 17 amino acids near the amino terminus (R41-V58) conserved in the rat and the human. This sequence has a higher nucleotidic match with the transcript variant 2 of the human GABAρ1 subunit. Xenopus oocytes injected with cRNA from the receptor generated currents when exposed to GABA that shared all the characteristics of other GABAρ1 subunits in mammals, including its modulation by dopamine. This study will help to increase the knowledge of the genetics of the pig, further the understanding of this important neurotransmitter receptor family and will shed some light in the evolution of these genes among mammals.
Department of Neurobiology and Behavior, University of California, Irvine, Irvine, CA 92697, USA. Electronic address: jreyesru@uci.edu.
Jorge Mauricio
JM
Reyes-Ruiz
Department of Neurobiology and Behavior, University of California, Irvine, Irvine, CA 92697, USA.
Agenor
A
Limon
Department of Neurobiology and Behavior, University of California, Irvine, Irvine, CA 92697, USA.
Ricardo
R
Miledi
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D017124:Conserved Sequence; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D051381:Rats; D018080:Receptors, GABA-B / Q000235:genetics / Q000378:metabolism*; D012160:Retina / Q000378:metabolism; D034421:Sus scrofa; D014981:Xenopus
D021122:Protein Subunits; D018080:Receptors, GABA-B
ESTs; Electrophysiology; Sequencing; Xenopus oocyte; mRNA