Functional beta2-adrenergic receptors are present in nonstrial tissues of the lateral wall in the gerbil cochlea.
Recently, we have demonstrated that functional beta1-adrenergic receptors are the dominant beta-adrenergic receptor subtype in the stria vascularis and that beta1-adrenergic receptors stimulate K+ secretion in strial marginal cells. The goal of the present study was to determine whether nonstrial tissues in the cochlear lateral wall contain beta-adrenergic receptors and if so which subtype is present. Pharmacological tools were used to identify receptors in functional studies where cAMP production was measured. Further, receptors were identified as transcripts by cloning and sequencing of reverse-transcriptase polymerase chain reaction (RT-PCR) products. Experiments were performed on gerbil nonstrial lateral wall tissues. Tissues stimulated with 10(-5) M isoproterenol produced 0.42 +/- 0.22 pmol cAMP per ear within 12 min (n = 14). The selective beta-adrenergic receptor agonist isoproterenol stimulated cAMP production with an EC50 of (2 +/- 3) x 10(-7) M (n = 7). Isoproterenol-stimulated cAMP production was inhibited by the beta2-adrenergic receptor antagonist ICI 118551 with an IC50 of (7 +/- 7) x 10(-6) M, which corresponds to an affinity constant of 1 x 10(-7) M (pK(DB) = 6.89 +/- 0.23, n = 3). Isoproterenol-stimulated cAMP production was not inhibited by the highly selective beta1-adrenergic receptor antagonist CGP 20712A. The IC50 and the affinity constant for CGP 20712A were estimated to be >3 x 10(-4) and >6 x 10(-6) M, respectively. RT-PCR of total RNA isolated from nonstrial lateral wall tissues using primers specific for beta1-, beta2- and beta3-adrenergic receptors revealed products of the predicted sizes for the beta1- and beta2- but not for the beta3-subtype. Sequence analysis confirmed that amplified cDNA fragments encoded gene-specific nucleotide sequences. These results demonstrate that nonstrial lateral wall tissues contain transcripts for beta1- and beta2- but not for beta3-adrenergic receptors and that the beta2-adrenergic receptor is the dominant functional receptor subtype. The cellular localization and function of the beta2-adrenergic receptors remains to be determined.
Schimanski
S
S
Cell Physiology Laboratory, Anatomy and Physiology Department, Kansas State University, Manhattan, Kans 66506, USA.
Scofield
M A
MA
Wangemann
P
P
D000319:Adrenergic beta-Antagonists / Q000494:pharmacology*; D000818:Animals; D003051:Cochlea / Q000187:drug effects*; D000242:Cyclic AMP / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D005849:Gerbillinae; D007093:Imidazoles / Q000494:pharmacology*; D007545:Isoproterenol / Q000494:pharmacology; D012313:RNA / Q000187:drug effects; D018343:Receptors, Adrenergic, beta-2 / Q000187:drug effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013316:Stria Vascularis / Q000187:drug effects
D000319:Adrenergic beta-Antagonists; D007093:Imidazoles; C034477:RNA primers; D018343:Receptors, Adrenergic, beta-2; D012313:RNA; C049763:CGP 20712A; D000242:Cyclic AMP; D007545:Isoproterenol