REGIONAL DIFFERENCES IN THE DENSITY AND SUBTYPE SPECIFICITY OF ENDOTHELIN RECEPTORS IN RABBIT URINARY-TRACT

We investigated the binding characteristics of endothelin (ET) receptors in rabbit ureter, bladder dome, bladder base, and urethra and compared the observed receptor properties with those of cloned human ET(A) and ET(B) receptors expressed in Chinese hamster ovary K-1 (CHO) cells. Receptor binding experiments with [I-125]ET-1 revealed the presence of a single class of specific, saturable, high affinity [I-125]ET-1 binding sites in all of the regions of the studied urinary tract. The rank order of the densities (B-max values) of [I-125]ET-1 binding sites was: ureter much greater than bladder dome > bladder base = urethra. ET-1 and ET-2 inhibited [I-125]ET-1 binding to the membrane particulates from the various regions of the urinary tract with single high affinity constants. A selective ET(A) receptor antagonist, BQ 123, and selective ET(B) agonists, ET-3 and sarafotoxin S6c (STXc), inhibited [I-125]ET-1 binding to bladder dome, bladder base, and urethra with high and low affinity constants indicating the presence of both ET(A) and ET(B) receptor subtypes in these tissues. The subtype specificity of ET receptors in the rabbit tissues is confirmed with inhibition data obtained from similar binding studies in cloned human ET(A) and ET(B) receptors. The proportions of high affinity binding sites for ET-3, representing ET(B) receptors, were approximately 25%, 27%, and 46% in bladder dome, bladder base, and urethra, respectively. Corresponding values for STXc were approximately 17%, 28%, and 43% in bladder dome, bladder base, and urethra, respectively. In contrast to the findings for ET-3 and STXc, the proportions of high affinity binding sites for BQ 123, representing ET(A) receptors, in bladder dome, bladder base, and urethra were approximately 84%, 74%, and 60%, respectively. In ureter, these selective compounds inhibited [I-125]ET-1 binding with either a low (ET-3 and STXc) or a high binding affinity (BQ 123), suggesting the presence of only a single receptor subtype (ET(A)) in this tissue. These data indicate that there are regional differences in the density and subtype specificity of ET receptors in the rabbit urinary tract.