Abstract,81-and fl2-adrenergic receptor (,B-ARs) expression in the thick ascending limb of rat kidney was studied at the level of mRNA and receptor coupling to adenylyl cyclase. Absolute quantitation off,l-and .82-AR mRNAs in microdissected nephron segments was performed with an assay based on reverse transcription and polymerase chain reaction, using in vitro transcribed mutant RNAs as internal standards. In the cortical thick ascending limb (CITAL), the number of mRNA molecules/mm of tubular length was 2,806±328 (n = 12) for,l1-AR and 159±26for,#2-AR (P < 0.01). Lower levels were obtained in the medullary thick ascending, fl1-AR mRNA still being predominant.The pharmacological properties of f-ARS was also studied in the CTAL. Cyclic AMP accumulation was stimulated by ,B-agonist with a rank order of potency of isoproterenol >norepineph-rine >epinephrine. This observation, and the higher efficiency of a 6i1 than of a f62 antagonist to inhibit isoproterenol-induced cAMP accumulation, establish the typical fl1-AR sensitivity of the CTAL. No detectable contribution of atypical or ,63-ARs to adenylyl cyclase stimulation could be found. In conclusion, this study, which shows markedly different levels of l1-and ,62-AR mRNAS in the CTAL, provides a molecular basis for the predominant expression of the ,l1 receptor subtype in this nephron segment. (J. Clin. Invest. 1993.91:264-272.)