Desensitization of ET(A) endothelin receptor-mediated negative chronotropic response in right atria--species difference and intracellular mechanisms.
1. Desensitization of ET(A) endothelin receptor (ET(A)R) was compared between the rat and guinea-pig with regard to negative chronotropic response (NC) in the right atria (RA). 2. ET-1 (100 nM) produced distinct NC in the presence of BQ788 (300 nM), and positive chronotropic response (PC) in the presence of BQ123 (1 microM) in both species, showing that ETAR and ET(B) endothelin receptor (ET(B)R) mediate NC and PC, respectively. 3. Repetitive applications of ET-1 (50 nM) desensitized PC, and the second application only induced a strong NC in both species. Later applications of ET-1 produced virtually no response in the rat RA, whereas they produced BQ123-sensitive NCs repetitively in guinea-pig RA, exhibiting marked species difference in desensitization of ETAR-mediated NC. 4. Pretreatment with staurosporine (100 nM) prevented desensitization of ET(A)R in the rat RA altogether. However, phorbol 12-myristate 13-acetate (PMA, 300 nM) failed to induce, but rather hampered, desensitization of ET(A)R. 5. Partial amino acid sequencing of ET(A)Rs, spanning from the 2nd through the 4th intracellular loops, revealed that all the potential Ser/Thr phosphorylation sites, including a protein kinase C (PKC) site, are conserved among guinea-pigs, rats, rabbits, bovines and humans. 6. In guinea pig RA, pretreatment with okadaic acid (1 microg ml(-1)) and PMA did not facilitate desensitization of ET(A)R whereas these agents successfully desensitized ETAR during combined stimulation of beta-adrenoceptor and ET(A)R by isoproterenol (300 nM) and ET-1 (100 nM). 7. These results suggest that species differences in desensitization of ET(A)R are not caused by differences in the site(s) of, but caused by differences in the environment for phosphorylation of the receptor. Desensitization of ET(A)R appears to require phosphorylation of the receptor by PKC as well as a kinase stimulated by beta-adrenoceptor activation.
Ono
K
K
Division of Chemical Pharmacology and Phytochemistry, National Institute of Health Sciences, Tokyo, Japan.
Sakamoto
A
A
Masaki
T
T
Satake
M
M
D000818:Animals; D001483:Base Sequence; D065128:Endothelin Receptor Antagonists; D006168:Guinea Pigs; D006325:Heart Atria / Q000187:drug effects; D006339:Heart Rate / Q000187:drug effects*; D008297:Male; D008969:Molecular Sequence Data; D019319:Okadaic Acid / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology*; D010456:Peptides, Cyclic / Q000494:pharmacology*; D010703:Phorbol Esters / Q000494:pharmacology; D010766:Phosphorylation; D010880:Piperidines / Q000494:pharmacology*; D011493:Protein Kinase C / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D011943:Receptors, Adrenergic, beta / Q000502:physiology*; D017466:Receptors, Endothelin / Q000187:drug effects* / Q000235:genetics; D013045:Species Specificity; D019311:Staurosporine / Q000494:pharmacology; D013997:Time Factors
D065128:Endothelin Receptor Antagonists; D009842:Oligopeptides; D010456:Peptides, Cyclic; D010703:Phorbol Esters; D010880:Piperidines; D011943:Receptors, Adrenergic, beta; D017466:Receptors, Endothelin; D019319:Okadaic Acid; C086539:BQ 788; D011493:Protein Kinase C; D019311:Staurosporine; C072247:cyclo(Trp-Asp-Pro-Val-Leu)