The rho(1) GABA receptor cloned from rat retina is down-modulated by protons

WE have cloned and sequenced the full-length cDNA of the rat GABA (gamma-aminobutyric acid) rho(1) receptor subunit. The deduced amino acid sequence (474 amino acids) of the rat rho(1) receptor is 95% homologous to the previously cloned human rho(1) receptor. The rho(1) cDNA includes a 5' 129 bp and a 3' 2.6 kb untranslated region, which contains a sequence homologous to the human medium reiteration frequency repetitive sequence, MER18. The rat rho(1) receptor shows 45-50% similarity to the GABA(A) receptor beta subunits. This similarity is among the highest between all GABA(A) receptor subunit classes, giving no support at the molecular level to the classification of the rho subunits to a novel GABA(C) receptor class. The rat rho(1) cDNA formed functional GABA receptors insensitive to bicuculline, but sensitive to blockade by picrotoxin, when transiently expressed in the human embryonic kidney cell line HEK 293. We studied the sensitivity of the rho(1)-mediated GABA current to variations in extracellular pH (pH(o)), and found that these receptors are strongly down-modulated by H+ ions. A decrease in pH(o) from 7.4 to 6.4 decreased the GABA current by 51 +/- 8%, whereas an increase in pH(o) from 7.4 to 8.4 increased the current by 77 +/- 8%. In view of the up-regulatory effect of protons on the GABA current observed in a number of preparations, the rho(1) receptors may contain a novel down-regulatory binding site for protons characteristic to these receptors.